analyte sheets-english - pribori.compribori.com/products/pdf/tosoh_reagent.pdf · the aia-pack b12...

SUMMARY AND EXPLANA-TION OF TEST Breast carcinoma-associated antigen coded by the human MUC-1gene is identified by several names including MAM 6, milk mucin, CA 27.29 and CA 15-3 (Fujirebio Diagnostics, Inc.). As indicated by epitope mapping, in-hibition of antibody binding, tracer exchange and clinical correlation studies, CA 27.29 is similar, if not identical, to CA 15-3. This antigen is a glycoprotein (Molecular weight 300 - 450 kDa), which contains 20-amino-acid tandem repetitive sequence of the mucin core and carbohydrate of more than half of the molecule (1). The number of tandem repeats and the degree of glycosylation is variable between individuals, so that CA 27.29 is very heterogeneous in structure. In malignant cells, CA 27.29 is overex-pressed on the entire cell surface, and increasing amounts are shed into the circulation. Tumors involving glandular organs, such as the breast, can produce high concentration of CA 27.29 in serum, making it useful as a tumour marker. Monoclonal antibodies that recog-nized distinct epitopes on CA 27.29 and CA 15-3 antigen molecules have been used to develop immunoassays (2-4). ST AIA-PACK 27.29 was devel-oped with a monoclonal antibody that recognized 8 amino acids in the tan-dem repeat portion of the polypeptide chain (5).Since the antigen levels cor-related closely with disease regres-sion or progression, CA27.29 proved reliable in the follow-up of patients with advanced breast cancer.(6,7)

PRINCIPLE OF THE ASSAY The ST AIA-PACK 27.29 is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. CA 27.29 present in the test sample is bound with monoclonal antibody immobilized on magnetic beads and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to re-move unbound materials and are then in-cubated with a fluorogenic substrate 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal an-tibody that binds to the beads is directly proportional to the CA 27.29 concentration in the test sample. A standard curve is constructed, and unknown sample con-centrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume: 20 µl Specimen type: Serum Assay range: up to 400 U/ml Calibration stability: 90 days Sensitivity: 0.10 U/ml

2277--2299

Reference range

<23 U/ml

Ver 05-01

PRECISION The intra-assay coefficient of variation was evaluated in two control samples by 10 repli-cate determinations. Sample Number of Mean Standard Coefficient Replicates Deviation of Variation ( U/mL) ( U/mL) (%) Control L 10 41.3 0.9 2.2 Control H 10 300.9 4.1 1.4 The inter-assay coefficient of variation was evaluated in two control samples by 21 repli-cate determinations. Sample Number of Mean Standard Coefficient Replicates Deviation of Variation ( U/mL) ( U/mL) (%) Control L 21 40.4 0.9 2.2 Control H 21 301.0 6.5 2.2 SENSITIVITY The minimal detectable concentration (MDC) of ST CA 27.29 is estimated to be 0.10 U/mL, which corresponds to 2 U/mL in original serum (dilution factor 21). The MDC is defined as that concentration of CA 27.29, which corresponds to the rate of fluorescence that is two Standard Deviations from the mean rate of fluorescence of 20 replicates of Calibrator (1). INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. - Hemoglobin (concentrations up to 412 mg/dL), free bilirubin (concentrations up to

16.5 mg/dL) and conjugated bilirubin (concentrations up to 18.2 mg/dL) do not inter-fere with the assay.

- Lipemia, as indicated by triglyceride (concentrations up to 1 667 mg/dL), does not interfere with the assay.

- Ascorbic acid (concentrations up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by albumin (concentrations up to 5 g/dL), does not interfere with

the assay.

ST AIA-PACK 27-29 025202 Calibrator Set 020302 Sample Diluting Solution 020502 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

.SUMMARY AND EXPLANA-TION OF TEST Alpha-fetoprotein (AFP) is a fetal globulin synthesized by the fetal liver, gastrointestinal tract and yolk sac(1) AFP is a glycoprotein with a molecular weight of 70,000 daltons containing 4.3% carbohydrate, of which approxi-mately 20% is sialic acid (2).Within a year after birth, AFP concentration in a normal serum decreases to a barely detectable level(3).Initially the test was used as an aid in the diagnosis of he-paticas(4), since increased serum AFP concentrations were first observed in some patients with primary hepatocel-lular carcinoma. Elevated serum AFP levels may also be observed in pa-tients with nonseminomatous testicular carcinoma (5,6,7).Increased serum AFP levels have not been observed in pa-tients with testicular carcinoma, which is of purely seminoma origin. In both nonseminomatous testicular carci-noma and primary hepatocellular car-cinoma in which AFP elevations were observed, the levels frequently decline during successful ther-apy(8,9,10).Because of this, serial moni-toring of serum AFP levels in such pa-tients has been shown to facilitate the clinical management of the disease. A few other common cancers, usually with hepatic metastases, increase AFP levels; these include carcinoma of stomach, gallbladder, prostate, and bronchi. Increased AFP levels have also been reported in non-malignant diseases including ataxia telangiec-tasia, hereditary tyrosinemia, neonatal hepatitis, atresia of the biliary tract (11,12,13,14) tumour of other tissues (15,16,17) as well as in pregnant women (18,19).

PRINCIPLE OF THE ASSAY The ST AIA-PACK AFP is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. AFP present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incu-bated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled mono-clonal antibody that binds to the beads is directly proportional to the AFP concen-tration in the test sample. A standard curve is constructed, and unknown sam-ple concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume: 25 µl Specimen type Serum Assay range: up to 100 ng/ml Calibration stability: 90 days Sensitivity: 1.0 ng/ml

Reference Interval

<10 ng/ml

AAFFPP

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 10.63 0.24 2.3 Serum B3 102.32 2.17 2.1 Serum C3 286.82 3.85 1.3 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 10.63 0.58 5.5 Serum B3 102.32 4.47 4.4 Serum C3 286.82 12.54 4.4 SENSITIVITY The minimal detectable concentration (MDC) of Alpha-Fetoprotein is estimated to be 1.0 ng/mL. The MDC is defined as that concentration of AFP, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. - Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17 mg/dL) and conjugated bilirubin (up to

18 mg/dL) do not interfere with the assay. - Lipemia, as indicated by triglyceride concentrations (up to 1666 mg/dL), does not interfere

with the assay. - Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not interfere with

the assay.

Chemotherapeutic Agents: Therapeutic concentrations of the following chemotherapeutic agents added to serum samples which were then assayed, do not interfere with ST AIA-PACK AFP Assay: Cisplatin, Velban, Bleomycin, Adriamycin, Vincristine, Methotrexate, 5-Fluorouracil, and Mitomycin C.

ST AIA-PACK AFP 025252 Calibrator Set 020352 Sample Diluting Solution 020552 Substrate Set II 020968 Wash Concentrate 020955

SUMMARY AND EXPLANATION OF TEST Synthesized exclusively by micro-organisms and found in nearly all animal tissues, vitamin B12 (cyanocobalamin) is essential for metabolism of fatty ac-ids, intermediary metabolism of folic acid, and the biosynthesis of DNA and RNA. After ingestion and initial protein digestion, the vitamin B12 forms com-plexes with the gastric glycoprotein car-riers, intrinsic factor (IF) and R factors. During digestion, the B12-IF complex is bound by receptors in the terminal ileal mucosa. The complex is dissociated as the vitamin B12 enters capillary to portal circulation for storage in the liver. Bound to the transport protein, transcobalamin II, vitamin B12 is released by hepato-cytes, as physiological needs arise. Deficiency of vitamin B12 may be pro-duced by any of several factors, includ-ing inadequate dietary intake and malabsorption. Malabsorption, the most common cause of vitamin B12 defi-ciency, is associated with auto-immune disorders (pernicious anemia), gastric disorders, and damage to or absence of the terminal ileum. Elevated levels of vitamin B12 are most commonly caused by ingestion of vita-min supplements. Elevated levels of vi-tamin B12, which persist after cessation of vitamin supplements, may indicate liver damage or myelo-poliferative dis-eases. Vitamin B12 deficiency can produce se-rious, progressive and if untreated, often irreversible neurological disorders and macrocytic or megaloblastic anemia. Because morphologically indistinguish-able megaloblastic anemias may be caused by deficiencies in either Vitamin B12 or folic acid, it is important to de-termine serum levels of both to properly diagnose the cause and effectively treat the anemia (1-6).

PRINCIPLE OF THE ASSAY The AIA-PACK B12 is a competitive enzyme immunoassay, which, after sample pre-treatment, is performed entirely within the AIA-PACK. Sample pre-treatment reagents (containing potassium cyanide, sodium hy-droxide, and dithiothreitol) release vitamin B12 from serum binding proteins in the sam-ple and converts cyanocobalamin into a sta-ble, measurable form of vitamin B12. Vitamin B12 present in the pre-treated test sample competes with enzyme-labelled vitamin B12 for a limited number of binding sites on a fluo-rescein labelled porcine intrinsic factor which then binds to anti-FITCH (fluorescein isothio-cyanate) antibody immobilized on magnetic beads. The beads are washed to remove the unbound enzyme labelled vitamin B12 and are then incubated with a fluorogenic sub-strate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme labelled vi-tamin B12 that binds to the beads is inversely proportional to the vitamin B12 concentrations in the test sample. A standard curve using a range of known standard concentrations is constructed and unknown vitamin B12 con-centrations are calculated using this curve.

Incubation time: 40 minutes Specimen volume: 100 µl Specimen type: Serum Assay range: up to 2000 pg/ml Calibration stability: 90 days Sensitivity: 84 pg/ml

Reference range

160 – 970 ng/ml

VViitt BB1122

Ver 05-01

PRECISION The infra-assay (within run) precision coefficient of variation was evaluated in four con-trol samples by 10 replicate determinations.

Sample Number of Mean Standard Coefficient Replicates Deviation of Variation (pg/mL) (pg/mL) % Control A 10 101 18.6 18.4 Control B 10 349 32.0 9.1 Control C 10 775 54.0 7.0 Control D 10 1210 75.8 6.3

The inter-assay (between run) precision coefficient of variation was evaluated at four dif-ferent concentrations by analysing controls in triplicate in 20 to 23 separate runs.

Sample Number of Mean Standard Coefficient Replicates Deviation of Variation (pg/mL) (pg/mL) % Control A 23 109 7.5 6.9 Control B 20 343 9.8 2.9 Control C 20 701 33.6 4.8 Control D 20 1311 59.9 4.6 SENSITIVITY The minimal detectable concentration (MDC) of vitamin B12 is estimated to be 84 pg/mL. The MDC is defined as that concentration of B12, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator (Calibrator 1). INTERFERENCE Interference is defined for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Added hemoglobin (up to 490 mg/dL), conjugated bilirubin (up to 14.3 mg/dL) and free bilirubin (up to 17.9 mg/dL) do not interfere with the assay.

- Lipemia, as indicated by added triglyceride (up to 1667 mg/dL), does not interfere with the assay.

- Protein, as indicated by added albumin (up to 2.5 g/dL), does not interfere with the assay.

- Ascorbic acid (up to 20.0 mg/dL) does not interfere with the assay.

ST AIA pack Vitamin B12 020293 Calibrator Set 020393 Sample Diluting Solution 020593 Pretreatment solution 020706 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Human chorionic gonadotropin (HCG) is a glycoprotein, which, like LH, FSH and TSH consists of alpha and beta chains. It contains approximately 30% carbohydrate by weight (1). The alpha chains are virtually identical in all four hormones, whereas the beta chains are different and determine both the specific biological activity and immuno-logical characteristics of each hormone (2,3). In the case of HCG, however, the amino acid sequence of the beta chain displays a considerable degree of ho-mology with that of LH(4), making it dif-ficult to achieve the required assay specificity based on conventional anti-body techniques. The use of mono-clonal antibody technology has made it possible to produce an essentially unlimited supply of antibody of pre-cisely defined specificity. HCG is produced by the placenta shortly after implantation(5-7) and in-creases at a geometric rate until it reaches a peak near the end of the first trimester. This makes it an excellent marker for the confirmation of preg-nancy and/or monitoring its course thereafter when necessary(5,8). Other conditions associated with elevated se-rum HCG concentrations are neo-plasms of trophoblastic and non-trophoblastic origin(9-12). Clinical applications of the HCG beta assay as a tumour marker include the diagnosis and management of hy-datidiform mole and chorio-carcinoma(13). The HCG beta assay is a valuable tool also in the diagnosis, staging and management of patients with testicular and ovarian germ-cell tumors(14). Ectopic production of HCG beta has been reported in cases such as carcinoma of the stomach, liver, pancreas, breast and in multiple mye-loma and melanoma(15).

PRINCIPLE OF THE ASSAY The ST AIA-PACK ß-HCG is a two-site immu-noenzymometric assay for the intact HCG molecule and beta subunit, which is performed entirely in the AIA-PACK. Intact HCG molecule and beta subunit present in the test sample are bound with monoclonal antibody immobi-lised on a magnetic solid phase, and then a distinctly different antigenic site on the beta subunits is bound with enzyme-labeled mono-clonal antibody in the AIA-PACK. The mag-netic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic substrate 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal anti-body that binds to the beads is directly propor-tional to the ß-HCG concentration in the test sample. A standard curve is constructed, and unknown sample concentrations are calcu-lated using this curve.

Incubation time: 10 minutes Specimen volume: 50 µl Specimen type: Serum Assay range: up to 400 mIU/ml Calibration stability: 90 days Sensitivity: 0.5 mIU/ml

Reference range Gestational Age mIU/mL

1 week 2 weeks 3 weeks 4 weeks

5 - 6 weeks 7 - 8 weeks

2 - 3 months 2nd trimester 3rd trimester

Non-pregnant/males

5 - 50 40 - 1.000 100 - 5.000 600 - 10.000 1.500 - 100.000 16.000 - 200.000 12.000 - 300.000 24.000 - 55.000 6.000 - 48.000 <5

ßß--HHCCGG

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (mIU/mL) (mIU/mL) (%) Serum A3 4.19 0.063 1.5 Serum B3 83.89 1.000 1.2 Serum C3 206.09 2.654 1.3 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (mIU/mL) (mIU/mL) (%) Serum A3 4.19 0.121 2.9 Serum B3 83.89 1.822 2.2 Serum C3 206.09 5.125 2.5 SENSITIVITY The minimal detectable concentration (MDC) of Total Beta HCG is estimated to be 0.5 mIU/mL. The MDC is defined as that concentration of ß-HCG, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. - Hemoglobin (up to 390.9 mg/dL), free bilirubin (up to 17.1 mg/dL) and conjugated

bilirubin (up to 17.7 mg/dL) do not interfere with the assay. - Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not

interfere with the assay. - Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not in-

terfere with the assay.

ST AIA-PACK B-HCG 025261 Calibrator Set 020361 Sample Diluting Solution 020561 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Human ß2 microglobulin (BMG) is a single polypeptide of 100 amino ac-ids, identified in 1968 by Berggard et al(1). The molecular weight of BMG is 11,800 daltons and represents the light-chain moiety of the major histo-compatibility complex class anti-gens(2-4).It is worth noting that BMG shows approximately 35% sequence homology with immuno–globulin do-mains. BMG is shed from the cell sur-face of all nucleated cells(2,3) during growth and differentiation, and cleared in the kidneys by filtration through the glomeruli. Since it is re-absorbed by the proximal tubular cells(5) and degraded, only trace amounts are detected in urine from healthy individuals, whereas marked elevation is seen in patients with proximal tubular dysfunctions due to exposure to aminoglycosides(6), anti-cancer or inflammatory drugs(7,8) and heavy metals(9). Determination of serum BMG is of diagnostic value in a variety of disor-ders. It can be used to assess glome-rular function in advanced renal dis-ease(10,11) and after renal trans-plantation(12,13).It can provide a measure of tumour burden and prog-nosis in patients with multiple mye-loma(14,15), B cell lymphoma(16) and chronic lymphocytic leukemia(17). It is useful to monitor disease activity in chronic inflammatory disorders, such as rheumatoid arthritis(18), systemic lupus erythematosis(19), Sjogren's syndrome(20), and Crohn's disease(21). More recently, BMG has been re-ported to be of value in monitoring patients with HIV infection(22-25).

PRINCIPLE OF THE ASSAY The ST AIA-PACK BMG is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. ß2 mi-croglobulin present in the test sample is bound with monoclonal antibody immobi-lized on a magnetic solid phase and en-zyme-labeled polyclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal an-tibody that binds to the beads is directly proportional to the BMG concentration in the test sample. A standard curve is con-structed, and unknown sample concentra-tions are calculated using this curve.

Incubation time: 10 minutes Specimen volume: 15 µl Specimen type: Serum/urine Assay range: up to 0.400 mg/l Calibration stability: 90 days Sensitivity: 0.002 mg/l

Reference Range

Serum 0.85 – 1.62 mg/L Urine < 0.3 mg/L

BBMMGG

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (mg/L) (mg/L) (%) Serum A3 0.023 0.00074 3.2 Serum B3 0.103 0.00250 2.4 Serum C3 0.247 0.00504 2.0 Urine A3 0.028 0.00047 1.7 Urine B3 0.227 0.00397 1.7 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean Pooled SD CV (mg/L) (mg/L) (%) Serum A3 0.023 0.00109 4.7 Serum B3 0.103 0.00386 3.7 Serum C3 0.247 0.00932 3.8 Urine A3 0.028 0.00087 3.1 Urine B3 0.227 0.00745 3.3 SENSITIVITY The minimal detectable concentration of ß2 microglobulin is estimated to be 0.002 mg/L as the "instrument measured concentration". For a 1:5 dilution of urine, this equates to 0.01 mg/L; for a 1:51 dilution of serum, this equates to a concentration of 0.10 mg/L. The minimal detectable con-centration is defined as that concentration of ß2 microglobulin, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17,1 mg/dL) and conjugated bilirubin (up to 17,7 mg/dL) do not interfere with the assay.

- Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not interfere with the assay.

- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not interfere

with the assay.

ST AIA-PACK BMG 025259 Calibrator Set 020359 Sample Diluting Solution 020559 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

CCEEAA SUMMARY AND EXPLANATION OF TEST Carcinoembryonic Antigen or CEA is a protein-polysaccharide b-globulin with a molecular weight of approximately 200,000 daltons and a sedimentation coefficient of 7S-8S(1,2,3). It is normally present at very low concentrations in the blood of healthy adults. Although elevated serum CEA levels were origi-nally thought to be specific for colorectal cancer(4,5), a variety of neoplastic and other disease processes cause signifi-cant elevation of CEA(6-13). Thus, CEA is used as a tumour marker in the man-agement of patients with colorectal, breast, lung, prostatic, pancreatic, ovar-ian and other carcinomas, as long as these tumors secrete the CEA moiety. Preoperative CEA levels may be of prognostic value since the level of ele-vation is correlated with body burden of tumour(8,14,15). Persistent elevation or increasing levels of CEA may indicate the presence of residual or dissemi-nated malignancy(16). Since CEA con-centration frequently declines during successful therapy and increases during disease progression, monitoring of se-rum CEA levels in patients with tumours which produce CEA has been shown to facilitate the clinical management of the disease. Measurement of serum CEA is not recommended as a screening pro-cedure since elevated CEA levels are also seen in a variety of non-neoplastic and benign diseases such as alcoholic cirrhosis, hepatitis, ulcerative colitis, and Crohn's disease(17,18) as well as in heavy cigarette smokers(19). Likewise, since not all tumors are CEA producing, a serum CEA measurement within the reference intervals found in apparently healthy subjects does not rule out the possibility of the presence of malignant disease.

PRINCIPLE OF THE ASSAY The ST AIA-PACK CEA is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. CEA present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to re-move unbound enzyme-labeled mono-clonal antibody and are then incubated with a fluorogenic substrate, 4-methylum-belliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly propor-tional to the CEA concentration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume: 100 µl Specimen type: Serum Assay range: up to 100 ng/ml Calibration stability: 90 days Sensitivity: 0.5 ng/ml

Reference range Non smoker <7 ng/ml Smoker <10 ng/ml

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 3.28 0.080 2.5 Serum B3 10.71 0.213 2.0 Serum C3 76.99 1.539 2.0 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 3.28 0.149 4.6 Serum B3 10.71 0.399 3.7 Serum C3 76.99 2.876 3.7 SENSITIVITY The minimal detectable concentration (MDC) of Carcinoembryonic Antigen is esti-mated to be 0.5 ng/mL. The MDC is defined as that concentration of CEA, which cor-responds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17 mg/dL) and conjugated bilirubin (up to 19 mg/dL) do not interfere with the assay.


- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Heparin (up to 100 U/ml) does not interfere with the assay.

ST AIA-PACK CEA 025254 Calibrator Set 020354 Sample Diluting Solution 020554 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Creatine kinase (CK) is an enzyme found in many hu-man tissues, including skeletal muscle, myocardium, and the brain. The molecule is a dimer and exists as three major isoenzyme structures, composed of either two M monomers (CK-MM), two B monomers (CK-BB), or one M and one B monomer (CK-MB). These dimers differ in molecular structure as well as tissue of primary source. CK-MM is primarily found in skeletal muscle. CK-BB is found predominantly in the brain and intes-tinal tissue, while CK-MB is seen in greatest concen-tration in cardiac muscle. All forms of CK function in the reversible phosphorylation of creatine(1,2). Accor-ding to the literature, increased serum concentration of each isoenzyme is associated with a particular disease state. Serum CK-MM is usually consistent with skeletal muscle disease (e.g. Duchenne's muscular dystrophy) or skeletal muscle trauma. Serum CK-BB elevation has been related to multiple conditions including Reye's syndrome, malignant hyperthermia, acute brain injury, cardiac failure, neurosurgical procedures and can be found even in some types of carcinoma such as gastric or prostatic. The MB isoenzyme is widely used in the diagnosis of acute myocardial infarction(3) CK activity in serum begins to increase 4-6 hours fol-lowing myocardial infarction, with peak activity reached between 18 and 30 hours. CK-MB isoenzyme returns to a normal level very rapidly, often within 24-36 hours. The diagnosis of acute myocardial infarction (AMI) is supported when serial serum samples from suspected patients show this characteristic elevation and then decline of CK-MB over the specified period of time (usually 24-36 hours). It is this temporal pattern rather than the abnormal elevation in CK-MB, which is indica-tive of the AMI. CK-MB can also be released from damaged cardiac cells in necrosis or trauma. Eleva-tions of CK-MB have also been reported in regenera-tive disorders(4). There are limitations to the use of an abnormal CK-MB as an indication of AMI, therefore, all pertinent informa-tion such as clinical history, ECG results, LD1 isoen-zyme abnormalities, ratio of CK-MB:total CK and the pattern of changes in CK-MB levels in serial samples should be taken into account when making the diagno-sis of AMI. Many methods are available for the separation and quantification of CK isoenzymes, including electro-phoresis, ion-exchange chromatography, immuno-inhibition, competitive immunoassays and immuno-metric methods. These procedures may measure ei-ther the enzymatic activity of the CK-MB isoenzyme or the mass concentration of the CK-MB fraction. The ST AIA-PACK CK-MB is a highly specific mass assay, which is not affected by high levels of the other CK isoenzymes.

PRINCIPLE OF THE ASSAY The ST AIA-PACK CK-MB is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. CK-MB present in the test sample is bound with mono-clonal antibody immobilized on a magnetic solid phase and enzyme-labelled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound en-zyme-labelled monoclonal antibody and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labelled monoclonal antibody that binds to the beads is directly propor-tional to the CK-MB concentration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 50 µl Specimen type: Serum Assay range up to 500 ng/ml Calibration stability: 90 days Sensitivity: 0.5 ng/ml

Reference range

<3.7 ng/mL

CCKK--MMBB

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 9.94 0.380 3.8 Serum B3 97.7 2.38 2.3 Serum C3 287.6 7.25 2.5 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 9.94 0.366 3.7 Serum B3 97.7 2.55 2.6 Serum C3 287.6 8.46 2.9 SENSITIVITY The minimal detectable concentration (MDC) of Creative Kinase MB Isoenzyme is esti-mated to be 0.5 ng/mL. The MDC is defined as that concentration of CK-MB, which cor-responds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. 1. Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17 mg/dL) and conjugated

bilirubin (up to 18 mg/dL) do not interfere with the assay. 2. Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not in-

terfere with the assay. 3. Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. 4. Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not inter-

fere with the assay.

ST AIA-PACK CK-MB 025269 Calibrator Set 020369 Sample Diluting Solution 020569 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

CCoorrttiissooll SUMMARY AND EXPLANATION OF TEST Cortisol, the major glucocorticoid pro-duced by the adrenal cortex regulates glucose, lipid and protein metabolism and has anti-inflammatory and immunosup-pressive effects (1,2). The cortisol level in circulation is regulated by adrenocortico-tropic hormone (ACTH) (3). ACTH is pro-duced by the pituitary gland and is regu-lated by the hypothalamus with cortico-tropin releasing factor(s) (CRF) (1). The cortisol itself also negatively regu-lates ACTH production. Therefore, meas-urement of serum cortisol and monitoring a circadian pattern (4,5) can provide infor-mation to evaluate hypothalamic-pituitary-adrenal function as well as clinical diag-nosis (6,7).

PRINCIPLE OF THE ASSAY The ST AIA-PACK CORT is a competi-tive enzyme immunoassay, which is performed entirely within the AIA-PACK. Cortisol present in the test sample com-petes with enzyme-labeled cortisol for a limited number of binding sites on a cor-tisol-specific antibody immobilized on magnetic beads. The beads are washed to remove the unbound enzyme labeled cortisol and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme labeled cortisol that binds to the beads is inversely pro-portional to the cortisol concentration in the test sample. A standard curve using a range of known standard concentra-tions is constructed and unknown corti-sol concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume: 10 µl Specimen type: Serum Assay range: up to 60 µg/dl Calibration stability: 90 days Sensitivity: 0.2 µg/dl

Reference range AM (8-10hrs) 8 - 25 µg/dl PM (16-18hrs) 1 - 17 µg/dl Midnight 1 – 5 µg/dl Urine <100 µg/24hrs

SPECIFICITY The following substances were tested for cross-reactivity. The cross-reactivity (%) is the percent of the compound, which will be identified as cortisol. If these compounds are present in the specimen at the same concentration as cortisol, the final result will be increased by these percentages. Compound Cross-reactivity Prednisolone 47.5 Cortisone 34.3 Prednisone 2.7 17a-Hydroxyprogesterone 0.28 Corticosterone 2.6 Testosterone < 0.001 Progesterone < 0.001 17b-estradiol < 0.001

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV ( µg/dL) ( µg/dL) (%) Serum A3 5.32 0.167 3.1 Serum B3 21.69 0.556 2.6 Serum C3 41.16 1.039 2.5 Total precision was determined by duplicate assays of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV ( µg/dL) ( µg/dL) (%) Serum A3 5.32 0.209 3.9 Serum B3 21.69 0.930 4.3 Serum C3 41.16 1.896 4.6 SENSITIVITY The minimal detectable concentration (MDC) of cortisol is estimated to be 0.2 µg/dL. The MDC is defined as that concentration of cortisol, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. - Hemoglobin (up to 436 mg/dL), free bilirubin (up to 17,7 mg/dL) and conjugated

bilirubin (up to 19,5 mg/dL) do not interfere with the assay. - Lipemia, as indicated by triglyceride concentrations (up to 1666 mg/dL), does not in-

terfere with the assay. - Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by added albumin (up to 5 g/dL), does not interfere with the as-

say.

ST AIA-PACK CORT 025287 Calibrator Set 020387 Sample Diluting Solution 020587 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST C-Peptide, a polypeptide 31 amino acids in length, originates in pancre-atic B-cells as a metabolically inert by-product in the synthesis of insulin from proinsulin(1). Insulin and C-Peptide are released from proinsulin in equimolar concentrations into the portal circulation(2). Therefore, C-Peptide levels can serve as an index to insulin secretion(3,4). Where insulin secretion is diminished, as in insulin-dependent diabetes, low C-Peptide levels are to be expected. Elevated C-Peptide levels may result from in-creased B-cell activity associated with insulinomas(4-6). Anti-insulin antibodies are commonly found in patients who have under-gone insulin therapy. These circulat-ing anti-insulin antibodies may inter-fere with certain immunoassays for insulin, making it difficult to use as a measure of residual B-cell activity. C-Peptide measurements are, there-fore, used as an alternative meas-urement index in this context(7). C-Peptide measurements are also used as an additional means of evaluating glucose tolerance tests(8).

PRINCIPLE OF THE ASSAY The ST AIA-PACK C-Peptide is a two-site immunoenzymometric assay, which is per-formed entirely in the AIA-PACK. C-Peptide present in the test sample is bound with antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to re-move unbound enzyme-labeled mono-clonal antibody and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal an-tibody that binds to the beads is directly proportional to the C-Peptide concentra-tion in the test sample. A standard curve is constructed, and unknown sample con-centrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 20 µl Specimen type: Serum/Urine Assay range: up to 30 ng/m Urine up to 300 ng/ml Calibration stability: 90 days Sensitivity: 0.2 ng/ml Urine 2.0 ng/ml

Reference range Serum: 1.1 － 3.3 ng/mL Serum concentrations typically increase post-prandially. Urine: 48 － 100 µg/24 hrs

CC--PPeeppttiiddee

SPECIFICITY Compound Cross-reactivity (%) C-Peptide 100 Proinsulin 69 Insulin 0.005

Ver 05-01

PRECISION Within run precision was determined using two controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 1.71 0.0478 2.8 Serum B3 5.23 0.1064 2.0 Total precision was determined by the duplicate assay of two controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (ng/mL) (ng/mL) (%) Serum A3 1.71 0.0966 5.7 Serum B3 5.23 0.3016 5.8 SENSITIVITY The minimal detectable concentration (MDC) of C-Peptide is estimated to be 0.2 ng/mL. The MDC is defined as that concentration of C-Peptide, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.



- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not

interfere with the assay.

ST AIA-PACK C-Pep 025284 Calibrator Set 020384 Sample Diluting Solution 020584 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Estradiol (1,3,5,10)-estratriene-3,17s-diol; E2) is a steroid hormone pro-duced mainly by the ovary in females and, in lesser amounts, by the testes in males (1). In non-pregnant subjects, there is cyclic variation in the concen-tration of E2 with the highest concen-trations occurring just prior to ovula-tion (2,3). In the first half of the men-strual cycle, follicle stimulating hor-mone (FSH) accelerates follicular de-velopment and E2 secretion. Rapid secretion of E2 imposes a positive feedback influence upon the pituitary gland, resulting in the release of lu-teinising hormone (LH) from the pitui-tary, and ovulation begins (4). The measurement of E2 simultane-ously with FSH and LH levels can provide information to evaluate ovary function (5). In males, E2 measure-ments are used for investigating fem-inizing syndromes such as gynaeco-mastia (6).

PRINCIPLE OF THE ASSAY The ST AIA-PACK E2 is a competitive en-zyme immunoassay, which is performed entirely within the AIA-PACK. Estradiol pre-sent in the test sample competes with en-zyme-labeled estradiol for a limited number of binding sites on a estradiol-specific anti-body immobilized on magnetic beads. The beads are washed to remove the unbound enzyme labeled estradiol and are then in-cubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme labeled estradiol that binds to the beads is inversely proportional to the estradiol concentration in the test sample. A standard curve using a range of known standard concentrations is con-structed and unknown estradiol concentra-tions are calculated using this curve.

Incubation time: 10 minutes Specimen vol 75 µl Specimen type: Serum Assay range: up to 3000 pg/ml Calibration stability: 90 days Sensitivity: 25 pg/ml

REFERENCE RANGE Category ranges (pg/mL) Male <75 Female Follicular phase 40.7 - 220.4 Ovulatory 209.0 - 424.6 Luteal phase 50.7 - 242.1 Post menopausal <75

EEssttrraaddiiooll

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (pg/mL) (pg/mL) (%) Serum A 110.2 6.70 6.1 Serum B 301.9 10.13 3.4 Serum C 747.3 19.72 2.6 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (pg/mL) (pg/mL) (%) Serum A 110.2 10.07 9.1 Serum B 301.9 16.67 5.5 Serum C 747.3 28.60 3.8 SENSITIVITY The minimal detectable concentration (MDC) of Estradiol is estimated to be 25 pg/mL. The MDC is defined as that concentration of E2, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 411 mg/dL), free bilirubin (up to 17,2 mg/dL) and conjugated bilirubin (up to 1,8 mg/dL) do not interfere with the assay.

- Lipemia, as indicated by triglyceride concentrations (up to 83 mg/dL) does not interfere with the assay.

- Protein, as indicated by added albumin (up to 1.2 g/dL), does not interfere with the assay.- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Conjugated bilirubin (more than 3.9 mg/mL) may interfere with the assay. Such speci-

mens may show falsely elevated E2 concentration.

ST AIA-PACK E2 025274 Calibrator Set 020374 Sample Diluting Solution 020574 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

FFeerrrriittiinnSUMMARY AND EXPLANATION OF THE TEST Ferritin is the major soluble iron storage protein from which iron may be mobi-lized for the synthesis of hemoglobin, Myoglobin and other iron-containing proteins(1-3). Ferritin is present in high concentra-tions in the cytoplasm of reticuloendo-thelial cells, liver cells, spleen cells and developing red cell precursors in bone marrow. Extracts from various tissues have different isoferritin distribution(3-6). The more acidic isoferritins are found primarily in heart, kidney and many tu-mour tissues. The liver, spleen and se-rum, on the other hand, contain the more basic isoferritins. Therefore, most immunoassays utilize basic isoferritins crystallized from human liver or spleen for the measurement of serum Ferritin. Studies on patients with iron-deficiency and iron overload abnormalities con-firmed that measurement of serum Fer-ritin concentrations reflect the amount of storage iron in the body(4-12).

PRINCIPLE OF THE ASSAY The ST AIA-PACK FER is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. Ferritin present in the test sample is bound with monoclonal antibody immo-bilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate, 4-methyl-umbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the Ferritin concentration in the test sample. A standard curve is con-structed, and unknown sample concen-trations are calculated using this curve.


Reference range Male 25.0 – 380 ng/ml Female 15.0 – 150 ng/ml

SPECIFICITY The cross-reactivity (%) is the percent of the compound, which will be identi-fied as Ferritin. If these compounds are present in the specimen at the same concentration as Ferritin, the final result will be increased by these percentages. Compound Cross-reactivity (%) Liver Ferritin 100 Spleen Ferritin 123.8 Heart Ferritin 5.6 Placental Ferritin 143.1

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 20.5 0.477 2.3 Serum B3 246.6 5.566 2.3 Serum C3 673.4 15.392 2.3 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard devia-tion (SD) and coefficient of variation (CV). Sample Mean SD CV (ng/mL) (ng/mL) (%) Serum A3 20.5 1.01 4.9 Serum B3 246.6 10.57 4.3 Serum C3 673.4 31.47 4.7 SENSITIVITY The minimal detectable concentration (MDC) of Ferritin is estimated to be 3.0 ng/mL. The MDC is defined as that concentration of FER, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 436 mg/dL), free bilirubin (up to 17.7 mg/dL) and conju-gated bilirubin (up to 19.5 mg/dL) do not interfere with the assay.


- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL),

does not interfere with the assay.

ST AIA-PACK FER 025253 Calibrator Set 020353 Sample Diluting Solution 020553 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

FFoollaatteeSUMMARY AND EXPLANATION OF TEST Synthesized exclusively by micro-organisms and found in nearly all animal tissues, vitamin B12 (cyanocobalamin) is essential for metabolism of fatty acids, intermediary metabolism of folic acid, and the biosynthesis of DNA and RNA. After ingestion and initial protein diges-tion, the vitamin B12 forms complexes with the gastric glycoprotein carriers, in-trinsic factor (IF) and R factors. During digestion, the B12-IF complex is bound by receptors in the terminal ileal mucosa. The complex is dissociated as the vita-min B12 enters capillary to portal circula-tion for storage in the liver. Bound to the transport protein, transcobalamin II, vi-tamin B12 is released by hepatocytes, as physiological needs arise. Deficiency of vitamin B12 may be pro-duced by any of several factors, including inadequate dietary intake and malab-sorption. Malabsorption, the most com-mon cause of vitamin B12 deficiency, is associated with auto-immune disorders (pernicious anemia), gastric disorders, and damage to or absence of the termi-nal ileum. Elevated levels of vitamin B12 are most commonly caused by ingestion of vitamin supplements. Elevated levels of vitamin B12, which persist after cessation of vi-tamin supplements, may indicate liver damage or myelopoliferative diseases. Vitamin B12 deficiency can produce seri-ous, progressive and if untreated, often irreversible neurological disorders and macrocytic or megaloblastic anemia. Be-cause morphologically indistinguishable megaloblastic anemias may be caused by deficiencies in either Vitamin B12 or folic acid, it is important to determine se-rum levels of both to properly diagnose the cause and effectively treat the ane-mia (1-6).

PRINCIPLE OF THE ASSAY The AIA-PACK B12 is a competitive enzyme immunoassay, which, after sample pre-treatment, is performed entirely within the AIA-PACK. Sample pre-treatment reagents (containing potassium cyanide, sodium hy-droxide, and dithiothreitol) release vitamin B12 from serum binding proteins in the sample and converts cyanocobalamin into a stable, measurable form of vitamin B12. Vi-tamin B12 present in the pre-treated test sample competes with enzyme-labelled vi-tamin B12 for a limited number of binding sites on a fluorescein labelled porcine intrin-sic factor which then binds to anti-FITCH (fluorescein isothiocyanate) antibody immo-bilized on magnetic beads. The beads are washed to remove the unbound enzyme la-belled vitamin B12 and are then incubated with a fluorogenic substrate, 4-methylum-belliferyl phosphate (4MUP). The amount of enzyme labelled vitamin B12 that binds to the beads is inversely proportional to the vi-tamin B12 concentrations in the test sample. A standard curve using a range of known standard concentrations is constructed and unknown vitamin B12 concentrations are calculated using this curve.


Reference range 2.0 – 16.0 ng/ml

Ver 05-01

PRECISION The infra-assay (within run) precision coefficient of variation was evaluated in four con-trol samples by 10 replicate determinations. Sample Number of Mean Standard Coefficient Replicates Deviation of Variation (pg/mL) (pg/mL) % Control A 10 101 18.6 18.4 Control B 10 349 32.0 9.1 Control C 10 775 54.0 7.0 Control D 10 1210 75.8 6.3 The inter-assay (between run) precision coefficient of variation was evaluated at four different concentrations by analysing controls in triplicate in 20 to 23 separate runs. Sample Number of Mean Standard Coefficient Replicates Deviation of Variation (pg/mL) (pg/mL) % Control A 23 109 7.5 6.9 Control B 20 343 9.8 2.9 Control C 20 701 33.6 4.8 Control D 20 1311 59.9 4.6 SENSITIVITY The minimal detectable concentration (MDC) of vitamin B12 is estimated to be 84 pg/mL. The MDC is defined as that concentration of B12, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator (Calibrator 1). INTERFERENCE Interference is defined for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.


- Lipemia, as indicated by added triglyceride (up to 1667 mg/dL), does not inter-fere with the assay.


- Ascorbic acid (up to 20.0 mg/dL) does not interfere with the assay.

AIA Vitamin Folate 020292 Calibrator Set 020392 Sample Diluting Solution 020592 Pretreatment solution 020707 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Follicle-Stimulating Hormone (FSH) is a glycoprotein, which, like LH, hCG and TSH consists of alpha and beta chains. The alpha chain is virtually identical in all four hormones, whereas the beta chains are different and determine both the specific bio-logical activity and immunological characteristics of each hormone(1,2). The FSH molecule contains approxi-mately 16% carbohydrate and has a molecular weight of approximately 28,000 to 30,000 daltons(3). FSH stimulates ovarian follicle growth and estrogen production in women, and testicular spermatogenesis in men(4-

7). Hypothalamic control of both FSH and LH secretion by the anterior pitui-tary appears to be by a common re-leasing hormone, gonadotropin-releasing hormone (GnRH) with negative feed back at the hypotha-lamic level by estrogen in the female and testosterone in the male. Determination of FSH concentrations is essential in assessment and moni-toring of patients with suspected in-fertility. Other physiological disorders associated with abnormal FSH secre-tion have been reported.

PRINCIPLE OF THE ASSAY The ST AIA-PACK FSH is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. FSH present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to re-move unbound enzyme-labeled mono-clonal antibody and are then incubated with a fluorogenic substrate 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal an-tibody that binds to the beads is directly proportional to the FSH concentration in the test sample. A standard curve is con-structed and unknown sample concentra-tions are calculated using this curve.

Incubation time: 10 minutes Specimen vol 50 µl Specimen type: Serum Assay range: up to 100 mIU/ml Calibration stability: 90 days Sensitivity: 1.0 mIU/ml

Reference range

Male 2.1 - 18.6 mIU/ml Ovulating Female

Follicular phase 4.5 - 11.0 mIU/ml Ovulatory 3.6 – 20.6 mIU/ml Luteal phase 1.5 – 10.8 mIU/ml

Postmenopausal Female 36.6 – 168.8 mIU/ml Boys 1.6 – 8.0 mIU/ml Girls 2.0 – 12.0 mIU/ml

FFSSHH

Ver 05-01

PRECISION

Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (mIU/mL) (mIU/mL) (%) Serum A3 4.96 0.128 2.6 Serum B3 16.37 0.290 1.8 Serum C3 60.15 0.886 1.5 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (mIU/mL) (mIU/mL) (%) Serum A3 4.96 0.276 5.6 Serum B3 16.37 0.774 4.7 Serum C3 60.15 2.573 4.3 SENSITIVITY The minimal detectable concentration (MDC) of Follicle-Stimulating Hormone is estimated to be 1.0 mIU/mL. The MDC is defined as that concentration of FSH, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17.7 mg/dL) and conjugated bilirubin (up to 18.2 mg/dL) do not interfere with the assay.




ST AIA-PACK FSH 025265 Calibrator Set 020365 Sample Diluting Solution 020565 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Triiodothyronine (T3) is present in human serum in an equilibrium mix-ture of bound and free forms, with approximately 0.4% of the total T3 circulating as Free Triiodothyronine (FT3). Any change in the serum con-centration of binding proteins will cause a parallel rise in the concentra-tion of total T3 with the FT3 remain-ing relatively unchanged (1). Its physiological action is apparent only when free fractions of T3 and T4 are taken in by target cell membrane re-ceptors (2,3,4). Since protein bound T3 is not available for cellular uptake and does not appear to exert any meta-bolic effect, the measurement of FT3 may therefore represent a more valid clinical assessment of patient thyroid status (5,6,7,8,9,10).Direct measurement of free T3 enables thyroid function examination even in the presence of abnormal liver function, hormone fluc-tuation during pregnancy (11), and variations in levels of serum binding proteins.

PRINCIPLE OF THE ASSAY The ST AIA-PACK FT3 is a competitive enzyme immunoassay, which is per-formed entirely within the AIA-PACK. Free fraction of T3-(FT3) present in the test sample competes with enzyme-labelled T3 for a limited number of bind-ing sites on an T3 -specific antibody im-mobilised on magnetic beads. The beads are washed to remove the unbound en-zyme-labelled T3 and are then incubated with a fluorogenic substrate. The amount of enzyme-labelled T3 that binds to the beads is inversely proportional to the FT3 concentration in the test sample. A stan-dard curve using a range of known stan-dard concentrations is constructed and unknown FT3 concentrations are calcu-lated using this curve.

Incubation time: 10 minutes Specimen vol 50 µl Specimen type: Serum Assay range: up to 25.0 pg/ml Calibration stability: 90 days Sensitivity: 0.70 pg/ml

Reference range

2.1 – 3.8 pg/ml

FFTT33

SPECIFICITY The following substances were tested for cross-reactivity. Cross-reactivity is expressed in terms of the percentage of the concentra-tion of each substance which will be meas-ured as FT3. Compound Cross-reactivity % L-Triiodothyronine (L-T3) 100 D-Triiodothyronine (D-T3) 125 L -Thyroxine (L-T3) 0.15 D-Thyroxine (D-T4) 0.07 3, 3', 5'-Triiodothyronine 0.3 3, 5-Diiodo-L-tyrosine <0.001

Ver 05-01

PRECISION The Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to ob-tain the pooled standard deviation (SD), which was then used to calculate the coefficient of varia-tion (CV).

Standard Coefficient Number Mean Deviation of Variation Sample of Factor (pg/ml) (pg/ml) (%) Serum A 20 2.65 0.124 4.7 Serum B 20 5.44 0.212 3.9 Serum C 20 13.57 0.321 2.4

Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV).

Standard Coefficient Number Mean Deviation of Variation Sample of Factor (pg/ml) (pg/ml) (%) serum A 20 2.65 0.183 6.9 serum B 20 5.44 0.262 4.8 Serum C 20 13.57 0.492 3.6 SENSITIVITY The minimal detectable concentration (MDC) of FT3 is estimated to be 0.7 pg/ml. The MDC is de-fined as that concentration of FT3, which corresponds to the rate of fluorescence that is two Stan-dard Deviations below from the mean rate of fluorescence of 20 replicates of a zero calibrator (Calibrator #1). INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 390.9 mg/dl), free bilirubin (up to 17.1 mg/dl) and conjugated bilirubin (up to 18.2 mg/dl) do not interfere with the assay.

- Ascorbic acid (up to 20 mg/dl) does not interfere with the assay. - Lipemia, as indicated by triglyceride concentrations (up to 416.7 mg/dl) does not interfere

with the assay. - Heparin (up to 100 U/ml) does not interfere with the assay. - Citrate trisodique (up to 2 mg/ml) does not interfere with the assay

ST AIA-PACK FT3 025286 Calibrator Set 020386 Sample Diluting Solution 020586 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST L-thyroxine (3, 5, 3-, 5-L-tetraiodothyronine (T4)) produced by the thyroid gland, circu-lates in the blood 99.97% bound to plasma proteins including thyroxine-binding globulin (TBG), thyroxine binding prealbumin (TBPA) and albumin(1,2). Approximately 0.03% of the total circulating thyroxine is unbound. This free T4 (FT4) is believed to be the physiologically active portion of the thyroxine, which stimulates the metabolism and controls, via the pituitary, the feedback system involving the release of TSH(3). His-torically, measurement of total serum T4 (bound + free) has been used to assess the clinical status of the thyroid gland(4,5). How-ever, this analysis is not diagnostically ac-curate when significant changes occur in the serum binding proteins(6,7). Alterations in TBG concentration, pregnancy, oral con-traceptives, estrogen therapy or drugs which alter the binding of thyroxine to the carrier proteins may cause corresponding changes in the total T4 when unbound free thyroxine levels remain relatively un-changed(8,9). Therefore, measurement of the free T4 (FT4) typically correlates more closely to the patient's actual thyroid status than the total (10).

PRINCIPLE OF THE ASSAY The ST AIA-PACK FT4 is a competi-tive enzyme immunoassay, which is performed entirely within the AIA-PACK. The thyroxine not bound to serum proteins (free T4) competes with enzyme-labeled T4 for a limited number of binding sites on a T4-specific antibody immobilized on magnetic beads. After incubation, the beads are washed to remove the un-bound enzyme-labeled T4 and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phos-phate (4MUP). The amount of en-zyme-labeled T4 that binds to the beads is inversely proportional to the free T4 concentration in the test sample. A standard curve using a range of known standard concentra-tions is constructed and unknown sample free T4 concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 10 µl Specimen type: Serum Assay range: up to 9.0 ng/dl Calibration stability: 90 days Sensitivity: 0.1ng/dl

Reference range

0.82 – 2.0 pg/ml

FFTT44

SPECIFICITY The following substances were tested for cross-reactivity. Cross-reactivity is ex-pressed in terms of the percentage of the concentration of each substance which will be measured as thyroxine. Compound Cross-reactivity(%) L-Thyroxine (L-T4) 100.0% D-Thyroxine (D-T4) 29.5 L-Triiodothyronine (L-T3) 1.1 D-Triiodothyronine (D-T3) 0.6 3, 3, 5-Triiodothyropropionic acid 1.8 3, 5-Diiodothyropropionic acid 0.13 3, 5-Diiodo-L-tyrosine < 0.1

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/dL) (ng/dL) (%) Serum A 0.56 0.02 4.0 Serum B 1.634 0.04 2.4 Serum C 4.27 0.09 2.2 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (ng/dL) (ng/dL) (%) Serum A 0.55 0.027 4.9 Serum B 1.62 0.056 3.5 Serum C 4.27 0.101 2.4 SENSITIVITY The minimal detectable concentration (MDC) of Free Thyroxine is estimated to be 0.10 ng/dL. The MDC is defined as that concentration of FT4, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.



- Protein, as indicated by human albumin concentrations (up to 40 mg/ml), does not interfere with the assay.

ST AIA-PACK FT4 025268 Calibrator Set 020368 Sample Diluting Solution 020568 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Human chorionic gonadotropin (HCG) is a glycoprotein, which, like LH, FSH and TSH consists of alpha and beta chains. It contains approximately 30% carbohy-drate by weight(1). The alpha chains are virtually identical in all four hormones, whereas the beta chains are different and determine both the specific biologi-cal activity and immunological character-istics of each hormone(2,3). In the case of HCG, however, the amino acid sequence of the beta chain displays a considerable degree of homology with that of LH(4), making it difficult to achieve the required assay specificity based on conventional antibody techniques. The use of mono-clonal antibody technology has made it possible to produce an essentially unlim-ited supply of antibody of precisely de-fined specificity. HCG is produced by the placenta shortly after implantation(5-7), and increases at a geometric rate until it reaches a peak near the end of the first trimester. This makes it an excellent marker for the con-firmation of pregnancy and/or monitoring its course thereafter when necessary(5,8). Other conditions associated with ele-vated serum HCG concentrations are neoplasms of trophoblastic and non- tro-phoblastic origin(9-12).

PRINCIPLE OF THE ASSAY The ST AIA-PACK HCG is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. HCG present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to re-move unbound enzyme-labeled mono-clonal antibody and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal an-tibody that binds to the beads is directly proportional to the HCG concentration in the test sample. A standard curve is con-structed, and unknown sample concentra-tions are calculated using this curve.


Reference Range

Gestational Age mIU/mL Gestational Age mIU/mL 1 week

2 weeks 3 weeks 4 weeks

5 - 6 weeks

5 to 50 40 to 1.000 100 to 5.000 600 to 10.000

1.500 to 100.000

7 - 8 weeks 2 - 3 months 2nd trimester 3rd trimester Non pregnant

16.000 to 200.000 12.000 to 300.000 24.000 to 55.000 6.000 to 48.000 <5.0

HHCCGG

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (mIU/mL) (mIU/mL) (%) Serum A3 5.61 0.112 2.0 Serum B3 109.39 2.023 1.9 Serum C3 274.16 4.833 1.8 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (mIU/mL) (mIU/mL) (%) Serum A3 5.61 0.177 3.2 Serum B3 109.39 3.603 3.3 Serum C3 274.16 9.564 3.5 SENSITIVITY The minimal detectable concentration (MDC) of Human Chorionic Gonadotropin is es-timated to be 0.5 mIU/mL. The MDC is defined as that concentration of HCG, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.





ST AIA-PACK HCG 025256 Calibrator Set 020356 Sample Diluting Solution 020556 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TESTHuman growth hormone, also known as HGH or somatotropin, is one of the main hormones pro-duced by the anterior portion of the pituitary gland. Unlike most hormones, HGH acts to di-rectly stimulate the target cells without the in-termediate involvement of a target endocrine organ. HGH promotes growth in soft tissue, car-tilage and bones, aids protein synthesis and greatly influences the metabolism of carbohy-drates and lipids. HGH is a polypeptide made up of 191 amino acids and has a molecular weight of 21,500 daltons. The half life of circulating human growth hormone is relatively short (20-30 minutes). The release of HGH from the pituitary is influenced by a variety of factors including chemical stimuli and the balance between growth hormone re-leasing factor (GHRF) and somatostatin (growth hormone release inhibiting hormone)(1,2). Sleep, stress, exercise, fasting, high protein meals, hy-poglycemia and certain amino acids such as L-dopa, glucagon, arginine and histamine all cause increases in HGH secretion. HGH release is inhibited by glucose, cortisol, free fatty acids and circulating HGH. The effect of hypersecretion of growth hormone depends on the age of the patient. Acromegaly, a progressive enlargement of the extremities, is caused by excess HGH in adults. In children, hypersecretion causes the phenomenon of ac-celerated growth with weakened bones and muscles termed gigantism. Hypersecretion can be caused by pituitary hyperplasia or tumors or by secondary factors such as a lack of release inhibiting factors or ectopic production of HGH by certain types of carcinoma(3-5). Hyposecretion of HGH may be caused by le-sions within the hypothalamus, an increase in the production of somatostatin, a lack of growth hormone releasing factor, pituitary tumors that destroy the cells producing HGH or even dam-age to the pituitary by external factors such as trauma or infection. Children deficient in HGH are said to have a condition known as pituitary dwarfism. Measuring random growth hormone is not usually meaningful. Screening tests for HGH deficiency, HGH hypersecretion and assess-ment of pituitary-hypothalamic function, utilize a variety of provocative tests (challenge tests). In this type of testing, serial blood samples are drawn and HGH response is measured against criteria for the specific testing protocol.

PRINCIPLE OF THE ASSAY The ST AIA-PACK HGH is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. Growth hormone present in the test sample is bound with monoclonal anti-body immobilized on a magnetic solid phase and enzyme-labeled mono-clonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic substrate, 4-methyl-umbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is di-rectly proportional to the HGH concen-tration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 75 µl Specimen type: Serum Assay range: up to 50 ng/ml Calibration stability: 90 days Sensitivity: 0.07 ng/ml

Reference range

<2.1 ng/ml

HHGGHH

Ver 05-01

PRECISION The intra-assay (within run) precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 2.60 0.048 1.9 Serum B3 7.87 0.100 1.3 Serum C3 25.33 0.227 0.9 Plasma A3 1.62 0.031 1.9 Plasma B3 6.06 0.100 1.6 Plasma C3 17.00 0.256 1.5 The inter-assay (between run) precision coefficient of variation was evaluated at three different concentrations, by analysing samples and heparinized plasma in 20 separate runs over 20 days. Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 2.60 0.093 3.6 Serum B3 7.87 0.204 2.6 Serum C3 25.33 0.442 1.7 Plasma A3 1.62 0.033 2.0 Plasma B3 6.06 0.093 1.5 Plasma C3 17.00 0.212 1.9 SENSITIVITY The minimal detectable concentration (MDC) of human growth hormone is estimated to be 0.07 ng/mL. The MDC is defined as that concentration of HGH, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluores-cence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17.7 mg/dL) and conjugated

bilirubin (up to 19.5 mg/dL) do not interfere with the assay. Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not in-

terfere with the assay. Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not inter-

fere with the assay.

ST AIA-PACK HGH 025266 Calibrator Set 020366 Sample Diluting Solution 020566 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Like the other immunoglobulin classes, IgE exhibits the typical four chain structure consisting of two light chains and two heavy chains. The two light chains and the variable portion of the heavy chain are common to all immunoglobulin classes. This portion of the molecule also contains the antigen recognition site. The IgE class specificity is contained within the constant region of the molecule, which is comprised of approximately three quarters of each heavy chain from the carboxyl-terminal end(1). IgE is capable of binding to recep-tors on mast cells and basophils. Through a series of complex and poorly understood reactions, an IgE-antigen complex is trig-gered to release certain vasoactive agents, such as histamine, into the blood stream producing the symptoms charac-teristic of "allergies"(2). Total serum IgE levels are of value in dif-ferentiating the allergic individual from non-allergic patients(3). Significantly ele-vated IgE concentrations are reported in allergic rhinitis, extrinsic asthma, urticaria and atopic eczema(4-7). Other conditions associated with elevated levels of IgE have also been reported(8-11).

PRINCIPLE OF THE ASSAY The ST AIA-PACK IgE II is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. Human IgE present in the test sample is bound with monoclonal antibody immo-bilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the IgE concentration in the test sample. A stan-dard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 10 µl Specimen type: Serum Assay range: 4000 IU/ml Calibration stability: 90 days Sensitivity: 3.0 IU/ml

Reference range <1 yr < 20 IU/ml 1-3 yrs < 30 IU/ml 3-4 yrs < 45 IU/ml 4-6 yrs < 100 IU/ml 7-8 yrs < 180 IU/ml >8 yrs < 150 IU/ml

IIgg EE IIII

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (IU/ml) (IU/ml) (%) Serum A3 44.1 0.935 2.1 Serum B3 262.8 6.24 2.4 Serum C3 1754.0 48.2 2.8 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (IU/ml) (IU/ml) (%) Serum A3 44.1 1.24 2.8 Serum B3 262.8 8.24 3.1 Serum C3 1754.0 54.6 3.1 SENSITIVITY The minimal detectable concentration (MDC) of Immunoglobulin E is estimated to be 3.0 IU/ml. The MDC is defined as that concentration of IgE, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. Hemoglobin (up to 430 mg/dL), free bilirubin (up to 17 mg/dL) and conjugated

bilirubin (up to 19 mg/dL) do not interfere with the assay. Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not

interfere with the assay. Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not in-


ST AIA-PACK IgE II 025295 Calibrator Set 020395 Sample Diluting Solution 020595 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Insulin, the antidiabetic hormone, is produced in the pancreatic B cell as(1,2) a large preproinsulin containing 109 amino acid residues with a mo-lecular weight of approximately 11,500. This peptide is rapidly con-verted by cleavage to proinsulin con-sisting of 86 amino acid residues with a molecular weight of approximately 9,000 and is stored within the B cell secretory granules(3). Equimolar quan-tities of insulin (51 amino acids with a molecular weight of approximately 6,000) and C-peptide (31 amino acids, molecular weight of approximately 3,000) are produced through prote-olytic cleavage and then secreted along with a small amount of proinsu-lin(4). Insulin exists in polymeric forms depending on the pH and zinc content. The monometric insulin molecule is composed of two polypeptide chains, "alpha" and "beta" which are con-nected by two interchain disulfide bridges of cystine(3). Insulin is degraded in most tissues and has a plasma half-life of 7 - 15 minutes in man. It is also rapidly and completely inactivated in the gastroin-testinal tract. Daily production of insu-lin in a healthy adult is 40 - 50 units. After binding to its specific receptors on target cell membranes, insulin acts as an anabolic and anticatabolic hor-mone, influencing the rates of carbo-hydrate, lipid, protein and electrolyte metabolism(5-7). Insulin release is stimulated by glu-cose. A failure to respond to this glu-cose stimulus may be one of the fun-damental defects in human diabetes(8-

10). The factors stimulating or inhibiting insulin release or the factors decreas-ing the tissue response to insulin have been well documented

PRINCIPLE OF THE ASSAY The ST AIA-PACK IRI is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. Insulin present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labelled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labelled monoclonal antibody and are then incu-bated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labelled mono-clonal antibody that binds to the beads is directly proportional to the IRI concentra-tion in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 50 µl Specimen type: Serum Assay range: up to 320 µU/ml Calibration stability: 90 days Sensitivity: 0.5 µU/ml

Reference range

<17 µU/ml

IInnssuulliinn

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV). Sample Mean Pooled SD CV ( µU/mL) ( µU/mL) (%) Serum A3 12.3 0.277 2.3 Serum B3 92.2 1.324 1.4 Serum C3 164.4 2.436 1.5 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV ( µU/mL) ( µU/mL) (%) Serum A3 12.3 0.559 4.6 Serum B3 92.2 2.425 2.6 Serum C3 164.4 4.669 2.8 SENSITIVITY The minimal detectable concentration (MDC) of insulin is estimated to be 0,5 µU/mL. The MDC is defined as that concentration of Insulin, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. Free bilirubin (up to 17,1 mg/dL) and conjugated bilirubin (up to 18.2 mg/dL) do not interfere

with the assay. Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not interfere

with the assay. Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not interfere with

the assay. Hemoglobin could interfere with the assay particularly at low range. Haemolysed samples should not be used for the AIA-PACK IRI assay because hemolysis

may falsely bring down the value due to Insulin Degrading Enzyme in red blood cells

ST AIA-PACK IRI 025260 Calibrator Set 020360 Sample Diluting Solution 020560 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Human Luteinising Hormone (LH) is a glyco-protein, which, like FSH, HCG and TSH con-sists of alpha and beta chains. The Alpha chain is virtually identical in all four hor-mones, whereas the beta chains are differ-ent and determine both the specific biologi-cal activity and immunological characteristics of each hormone(1,2). The LH molecule contains 15-18% carbohy-drate and has a molecular weight of ap-proximately 29,000 daltons(3,4). LH induces ovulation and thereafter maintains the cor-pus luteum and progesterone production in women(5-8). In men, LH stimulates testoster-one production by the Leydig cells(5,6,9). Hypothalamic control of both LH and FSH secretion by the anterior pituitary appears to be by a common releasing hormone, gonad-otropin-releasing hormone (GnRH) or lu-teinising hormone-releasing hormone (LHRH), with negative feedback at the hypo-thalamic level by estrogen in female and tes-tosterone in male. Through the use of monoclonal antibody technology, which provides the necessary specificity and sensitivity, determination of LH concentrations is critically important in assessment and monitoring of patients with suspected infertility(10,11). The raised levels of LH and LH: FSH ratio is frequently found in polycystic ovary syndrome (PCOS), a com-mon cause of infertility in women(12,13) and other physiological disorders associated with abnormal LH secretion (12-15).

PRINCIPLE OF THE ASSAY The ST AIA-PACK LH II is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. LH present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled mono-clonal antibody in the AIA-PACK. The magnetic beads are washed to re-move unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly propor-tional to the LH concentration in the test sample. A standard curve is con-structed, and unknown sample con-centrations are calculated using this curve.


Reference range Male 1.7 – 11.2 mIU/mL Ovulating Female:

Follicular phase 1.7 – 13.3 mIU/mL Ovulatory 4.1 – 68.7 mIU/mL Luteal phase 0.5 – 19.8 mIU/mL

Postmenopausal 14.4 – 62.2 mIU/mL Boys 0.5 – 4.0 mIU/mL Girls 2.0 – 12.0 mIU/mL

LLHH IIII

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (mIU/mL) (mIU/mL) (%) Serum A3 4.94 0.123 2.5 Serum B3 25.27 0.597 2.4 Serum C3 98.14 1.733 1.8 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (mIU/mL) (mIU/mL) (%) Serum A3 4.94 0.134 2.7 Serum B3 25.27 0.535 2.1 Serum C3 98.14 2.032 2.1 SENSITIVITY The minimal detectable concentration (MDC) of Luteinising Hormone is estimated to be 0.2 mIU/mL. The MDC is defined as that concentration of LH, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. Hemoglobin (up to 411.8 mg/dL), free bilirubin (up to 17.1 mg/dL) and conjugated

bilirubin (up to 18.2 mg/dL) do not interfere with the assay. Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not



ST AIA-PACK LH II 025296 Calibrator Set 020396 Sample Diluting Solution 020596 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANA-TION OF TEST Myoglobin is an oxygen-binding heme protein of low molecular weight (17,800) that is found in both cardiac and skeletal muscle. Myoglobin is re-leased from the muscle cells and ap-pears in the circulation after injury to the skeletal muscles or myocardium (e.g. acute myocardial infarction or AMI), surgery, exercise, or myopathies.1 In AMI, damaged myocardial tissue leads to elevated Myoglobin above normal levels within 2-3 hours after onset of symptoms, reaching peak levels at 6 to 9 hours, and returning to the normal range by 24 hours.2 This rapid elevation of Myoglobin in the cir-culation has lead to its usefulness for the early diagnosis and monitoring of AMI. 3-5 Increases in Myoglobin con-centration following thrombolytic treatment act as an early predictor of infarct reperfusion.6-8 Successful reperfusion of a blocked coronary ar-tery is indicated by Myoglobin levels which peak between 30 minutes and 2 hours following thrombolytic therapy.

PRINCIPLE OF THE ASSAY The AIA-PACK Myoglobin is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. Myoglobin present in the test sample is bound with monoclonal antibody im-mobilised on a magnetic solid phase and enzyme-labelled monoclonal anti-body in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labelled monoclonal antibody and are then incubated with a fluoro-genic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of en-zyme-labelled monoclonal antibody that binds to the beads is directly pro-portional to the Myoglobin concentra-tion in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen vol 10 µl Specimen type: Serum Assay range: up to 1000 ng/ml Calibration stability: 90 days Sensitivity: 2.0 ng/ml

Reference range

<73.0 ng/mL

MMyyoogglloobbiinn

SPECIFICITY The following substances were tested for cross-reactivity. The cross-reactivity (%) is the percent of the compound, which will be identified as Myoglobin. If these compounds are present in the specimen at the same concentration as Myoglobin, the final result will be increased by these percentages. Compound Cross-reactivity (%) Haemoglobin 0.0

Ver 05-01

PRECISION The intra-assay (within run) precision coefficient of variation was evaluated in three control samples by 10 replicate determinations.

Sample

Number of Replicates

Mean

(ng/mL)

Standard Deviation (ng/mL)

Coefficient of Variation

(%) Sample A Sample B Sample C

10 10 10

32.6 110.0 930.0

0.52 3.04

26.20

1.6 2.8 2.8

The inter-assay (between run) precision coefficient of variation was evaluated at three different concentrations by analysing control samples in 30 separate runs.

Sample


Mean

(ng/mL)

Standard Deviation (ng/mL)


(%) Sample A Sample B Sample C

30 30 30

31.4 113.0 971.0

1.40 4.65

43.30

4.4 4.1 4.5

SENSITIVITY The minimal detectable concentration (MDC) of Myoglobin is estimated to be 2.0 ng/mL. The MDC is defined as that concentration of Myoglobin, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluores-cence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.


- Lipemia, as indicated by added triglyceride (up to 1000 mg/dL) does not interfere with the assay.

- Protein, as indicated by added albumin (up to 5 g/dL), does not interfere with the assay.


ST AIA-PACK MYO 025297 Calibrator Set 020397 Sample Diluting Solution 020597 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST CA125 determinant, identified in 1981 by Bast et al, is associated with a high mo-lecular weight (220K Dalton) glycopro-tein complex. The precise immunological and physiological function of the glyco-protein complex expressing CA125 is unknown (1). Elevated serum levels of CA125 are associated with greater than 80 percent of nonmucinous epithelial ovarian neoplasms. Other gynecological neoplasms primarily endometrial and fallopian tube carcinomas and certain tumors of the pancreas, liver, colon, breast, and lung are also associated with elevated serum levels of CA125 (2). Recent studies indicate that monitoring serial determinations of CA125 in con-junction with chemotherapy may have clinical value in deciding patient progno-sis (3). Studies also suggest that long term monitoring of CA125 in patients with known ovarian carcinomas has util-ity in detecting early recurrence and dis-ease progression (4).

SUMMARY AND EXPLANATION OF TEST CA125 determinant, identified in 1981 by Bast et al, is associated with a high mo-lecular weight (220K Dalton) glycopro-tein complex. The precise immunological and physiological function of the glyco-protein complex expressing CA125 is unknown (1). Elevated serum levels of CA125 are associated with greater than 80 percent of nonmucinous epithelial ovarian neoplasms. Other gynecological neoplasms primarily endometrial and fallopian tube carcinomas and certain tumors of the pancreas, liver, colon, breast, and lung are also associated with elevated serum levels of CA125 (2). Recent studies indicate that monitoring serial determinations of CA125 in con-junction with chemotherapy may have clinical value in deciding patient progno-sis (3). Studies also suggest that long term monitoring of CA125 in patients with known ovarian carcinomas has util-ity in detecting early recurrence and dis-ease progression (4).

Incubation time: 10 minutes Specimen volume 100 µl Specimen type: Serum Assay range: up to 1000 U/ml Calibration stability: 90 days Sensitivity: 2.0 U/ml

Reference range

< 37 U/mL

OOVVCCAA

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV ( U/mL) ( U/mL) (%) Serum A3 21.5 0.701 3.3 Serum B3 102.5 2.64 2.6 Serum C3 657.6 22.8 3.5 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV ( U/mL) ( U/mL) (%) Serum A4 59.9 2.2 3.7 Serum B4 756 24.3 3.2 SENSITIVITY The minimal detectable concentration (MDC) of CA125 is estimated to be 2 U/mL. The MDC is defined as that concentration of CA125 which corresponds to the rate of fluo-rescence that is two standard deviations from the estimated mean rate of fluorescence of 10 replicate determinations of a zero calibrator (Calibrator (1)). INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.





ST AIA-PACK OVCA 025288 Calibrator Set 020388 Sample Diluting Solution 020588 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST The first acid phosphatase was discov-ered in 1924 and was found to be of erythrocyte origin(1). Prostatic acid phos-phatase (PAP) enzymatic activity was first measured on the urine of men in 1935(2) and was found to be localized in organs of the male genital tract(3,4). Since serum PAP concentrations were found to be elevated in many men with primary prostatic carcinoma and metas-tatic lesions of the prostate(5,6) it was suggested that PAP may be a significant tumour marker. Prostatic cancer is the second most common type of cancer found in men, occurring in approximately 50% of those over 70 years old(7,8). PAP is released into the blood stream by pri-mary tumors or following relapse of a prostatic malignancy(9-11). Therefore, sensitive and accurate measurement of serum PAP is essential in monitoring the effectiveness of therapy(9-14).

PRINCIPLE OF THE ASSAY The ST AIA-PACK PAP is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. PAP present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incu-bated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled mono-clonal antibody that binds to the beads is directly proportional to the PAP concen-tration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume 75 µl Specimen type: Serum Assay range: up to 40 ng/ml Calibration stability: 90 days Sensitivity: 0.2 ng/ml

Reference range

<1.2 ng/mL

PPAAPP

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 2.13 0.042 2.0 Serum B3 5.34 0.100 1.9 Serum C3 31.95 0.630 2.0 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (ng/mL) (ng/mL) (%) Serum A3 2.13 0.073 3.4 Serum B3 5.34 0.187 3.5 Serum C3 31.95 1.079 3.4 SENSITIVITY The minimal detectable concentration (MDC) of Prostatic Acid Phosphatase is esti-mated to be 0.2 ng/mL. The MDC is defined as that concentration of PAP, which corre-sponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. - Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17 mg/dL) and conjugated

bilirubin (up to 18 mg/dL) do not interfere with the assay. - Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not

interfere with the assay. - Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not in-


ST AIA-PACK PAP 025262 Calibrator Set 020362 Sample Diluting Solution 020562 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Progesterone (pregn-4-ene-3, 20-dione) is a steroid hormone produced mainly by the ovary and adrenal cortex in females and, in lesser amounts, by the adrenal cortex in males (1,2). Progesterone is se-creted in small amounts by the ovaries during the follicular phase of the men-strual cycle and increases sharply follow-ing ovulation and corpus luteum devel-opment (3). Unless pregnancy occurs, a steep decline to follicular levels occurs prior to the next menstrual cycle (4). This pattern provides the rationale for the use of serum progesterone measurements as a reliable method for ovulation detec-tion(5). Daily progesterone levels are considered the most accurate means for document-ing luteal phase defect (6). Measurements of serum progesterone can also be used to detect and evaluate patients at risk for abortion during the early weeks of gesta-tion (7,8).

PRINCIPLE OF THE ASSAY The ST AIA-PACK PROG is a competi-tive enzyme immunoassay, which is per-formed entirely within the AIA-PACK. Progesterone present in the test sample competes with enzyme-labeled proges-terone for a limited number of binding sites on a progesterone-specific anti-body immobilized on magnetic beads. The beads are washed to remove the unbound enzyme labeled progesterone and are then incubated with a fluoro-genic substrate, 4-methyl-umbelliferyl phosphate (4MUP). The amount of en-zyme labeled progesterone that binds to the beads is inversely proportional to the progesterone concentration in the test sample. A standard curve using a range of known standard concentrations is constructed and unknown progesterone concentrations are calculated using this curve.


Reference range Male < 0.5 Female Follicular phase < 1.0 Luteal phase 0.37 ~ 18.4 Post menopausal < 0.5

PPrrooggeesstteerroonnee

SPECIFICITY The following substances were tested for cross-reactivity. Cross-reactivity (%) is the percent of the compound, which will be iden-tified as progesterone. If these compounds are present in the specimen at the same concentration as progesterone, the final re-sult will be increased by these percentages. Compound Cross-reactivity Progesterone 100.0 17a-Hydroxyprogesterone 1.2 11-Deoxycorticosterone 3.5 Pregnelone 0.3 Corticosterone 1.79 5b-Pregnan-3,20-dione 6.3 Testosterone 0.03 Cortisol ND 5a-Pregnan-3,20-dione 12.8

Ver 05-01

PRECISION Within run precision was determined using four controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coefficient of variation (CV).

Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Control A 0.902 0.0962 10.7 Control B 2.48 0.158 6.4 Control C 8.65 0.437 5.1 Control D 17.73 0.577 3.3 Total precision was determined by the duplicate assay of four controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV).

Sample Mean SD CV (ng/mL) (ng/mL) (%) Control A 0.902 0.117 12.9 Control B 2.48 0.205 8.3 Control C 8.65 0.541 6.3 Control D 17.73 0.826 4.7 SENSITIVITY The minimal detectable concentration (MDC) of progesterone is estimated to be 0.1 ng/mL. The MDC is defined as that concentration of progesterone, which corresponds to the rate of fluores-cence that is two standard deviations from the mean rate of fluorescence of 20 replicate determina-tions of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 412 mg/dL), free bilirubin (up to 15.5 mg/dL) and conjugated bilirubin (up to 1.8 mg/dL) do not interfere with the assay.

- Lipemia, as indicated by triglyceride concentrations (up to 71 mg/dL) does not inter-fere with the assay.


- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Conjugated bilirubin (more than 9.5 mg/mL) may interfere with the assay. Such

specimens may show falsely elevated PROG concentration.

ST AIA-PACK PROG 025281 Calibrator Set 020381 Sample Diluting Solution 020581 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Prolactin (PRL) is a protein of 198 amino acids with a molecular weight of ap-proximately 21,000. Prolactin is secreted from the anterior pituitary gland and acts directly on the mammary gland to stimu-late milk production(1,2). When secreted in excess, it increases adrenal andro-gens and blunts the response of gonad-otropes and gonadal cells to tropic fac-tors by decreasing gonadoliberin. Thus, chronically elevated Prolactin induces amenorrhea and infertility in women (3-5) and impotence in men. Prolactin synthe-sis and secretion is under inhibitory con-trol by the hypothalamus. The hypotha-lamic Prolactin Inhibiting Factor (PIF) is now established as dopamine(1,2,6). Measurement of serum Prolactin con-centrations is essential in evaluating pa-tients with infertility or suspected hypo-thalamic-pituitary dysfunction. Increased serum Prolactin concentrations are ob-served in pregnancy and during lacta-tion. Other conditions associated with abnormal Prolactin concentrations have been reported(3,6,7-11).

PRINCIPLE OF THE ASSAY The ST AIA-PACK PRL is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. Prolactin present in the test sample is bound with monoclonal antibody immo-bilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the Prolactin concentration in the test sam-ple. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.


Reference range

Male 3.6 – 16.3 ng/mL Female 4.1 – 28.9 ng/mL

PPrroollaaccttiinn

SPECIFICITY The following substances were tested for cross-reactivity. The cross-reactivity (%) is the percent of the compound, which will be identified as Prolactin. If these compounds are present in the specimen at the same concentration as Prolactin, the final result will be increased by these percentages. Compound Cross-reactivity (%) PRL 100 LH 0.03 FSH < 0.01 TSH < 0.01 hPL < 0.01 hGH 0.04

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 4.32 0.0898 2.1 Serum B3 20.6 0.299 1.5 Serum C3 100.7 1.77 1.8 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV).

Sample Mean SD CV (ng/mL) (ng/mL) (%) Serum A3 4.32 0.126 2.9 Serum B3 20.6 0.580 2.8 Serum C3 100.7 2.72 2.7 SENSITIVITY The minimal detectable concentration (MDC) of Prolactin is estimated to be 1.0 ng/mL. The MDC is defined as that concentration of PRL, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. Hemoglobin (up to 390.9 mg/dL), free bilirubin (up to 17.1 mg/dL) and conjugated

bilirubin (up to 18.2 mg/dL) do not interfere with the assay. Lipemia, as indicated by triglyceride concentrations (up to 1660 mg/dL), does not



ST AIA-PACK PRL 025255 Calibrator Set 020355 Sample Diluting Solution 020555 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Prostate Specific Antigen (PSA) was identified in 1977 by Wang, et al.1 PSA is a single chain glycoprotein of ap-proximately 34,000 daltons containing 7% carbohydrate.2 Functionally and im-munohistochemically, PSA is distinct from prostatic acid phosphatase (PAP)3 and is confined to the cytoplasm of prostatic acinar cells and ductal epithe-lium.4 The presence of PSA has been demon-strated in normal, benign hyperplastic, and malignant prostatic tissue, in metas-tatic prostatic carcinoma, and also in prostatic fluid as well as seminal plasma.5 PSA is not present, however, in any other tissue from men, nor is it produced by cancers of the lung, colon, rectum, stomach, pancreas or thyroid.6 Elevated serum PSA levels have been reported in patients with prostate cancer, benign prostatic hypertrophy, or inflam-matory conditions of other adjacent geni-tourinary tissues.3,7 Prostate cancer cannot be diagnosed until biopsy results confirm the presence of cancer cells. Studies indicate that PSA is an impor-tant tool in assessing the effect of ther-apy.8 Especially in patients being treated with hormone therapy, concurrent serial determinations of PSA and PAP may provide added clinical value in monitor-ing patients with prostatic cancer.9

PRINCIPLE OF THE ASSAY The ST AIA-PACK PA is a two-site im-munoenzymometric assay, which is per-formed entirely in the ST AIA-PACK. PSA present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the ST AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the PSA concentration in the test sample. A stan-dard curve is constructed, and unknown sample concentrations are calculated using this curve.


Reference range

<49 yrs < 2.0 ng/ml 50 – 59 yrs < 3.5 ng/ml 60 – 69 yrs < 4.5 ng/ml 70 – 79 yrs < 6.5 ng/ml

PPSSAA

Ver 05-01

Precision

Within run precision was determined using three controls in total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation(SD), which was then used to calcu-late the coefficient of variation(CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A 2.27 0.055 2.4 Serum B 7.08 0.110 1.6 Serum C 29.54 0.487 1.6 The total precision was determined by the duplicate assay of three controls in 20 sepa-rate runs. The means of each run were used to calculate the standard deviation(SD) and coefficient of variation(CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A 2.27 0.094 4.1 Serum B 7.08 0.209 3.0 Serum C 29.54 0.934 3.2 Sensitivity The minimal detectable concentration (MDC) of Prostate Specific Antigen is estimated to be 0.1 ng/mL. The MDC is defined as that concentration of PSA, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluo-rescence of 10 replicate determinations of a zero calibrator. Interference Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 390 mg/dL) , free bilirubin(up to 16 mg/mL) and conjugated bilirubin (up to 18 mg/dL) do not interfere with the assay.

- Lipemia, as indicated by triglyceride concentrations(up to 1,660 mg/dL), does not interfere with the assay.

- Ascorbic acid (up to 20mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 5 g/dL), does not


ST AIA-PACK PA 025263 Calibrator Set 020363 Sample Diluting Solution 020563 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Carbohydrate antigen 19-9 (CA 19-9) is a digestive cancer associated antigen found by Koprowski and his co-workers in 1979(1,2). CA 19-9 is an antigen hold-ing sialyl Lewis A glycated chain as an antigen determinant and is present in blood in the form of mucin glycoprotein(3-

5). CA 19-9 is contained in tissues such as pancreas, gallbladder, meconium, and saliva as well as in tumour tissues. Elevated serum CA 19-9 levels have been reported in patients with pancreatic cancer, biliary cancer, carcinoma of co-lon and rectum, and liver cancer. Many of these examples are observed in pan-creatic and biliary cancers(6-8). Since false positive rate is less for be-nign diseases, measurement of CA 19-9 is widely used as an effective test for di-agnosis of malignant tumors of pancreas and biliary and as effective tools to make a decision of medical treatment effec-tiveness and to do follow-ups.

PRINCIPLE OF THE ASSAY The ST AIA-PACK SLa is a two-site im-munoenzymometric assay, which is per-formed entirely in the AIA-PACK. CA 19-9 present in the test sample is bound with monoclonal antibody immobilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incu-bated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled mono-clonal antibody that binds to the beads is directly proportional to the CA 19-9 con-centration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume 30 µl Specimen type: Serum Assay range: up to 400 U/ml Calibration stability: 90 days Sensitivity: 0.1 U/ml

Reference range

< 37 U/ml

SSLLaa

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV).

Sample Mean Pooled SD CV ( U/mL) ( U/mL) (%) Serum A3 20.5 0.572 2.8 Serum B3 30.3 0.766 2.5 Serum C3 59.2 1.349 2.3 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV ( U/mL) ( U/mL) (%) Serum A3 20.5 0.897 4.4 Serum B3 30.3 1.202 4.0 Serum C3 59.2 2.330 3.9 SENSITIVITY The minimal detectable concentration (MDC) of CA 19-9 is estimated to be 1 U/mL. The MDC is defined as that concentration of CA 19-9 which corresponds to the rate of fluo-rescence that is two Standard Deviations from the estimated mean rate of fluorescence of 10 replicate determinations of a zero calibrator (Calibrator 1). INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.



- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay. - Protein, as indicated by human albumin concentrations (up to 2.5 g/dL), does not


ST AIA-PACK SLa 025271 Calibrator Set 020371 Sample Diluting Solution 020571 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

Summary and Explanation of Test Testosterone is one of major male sex hormone produced by the interstitial cells of Leydig in the testes. The measurement of total testosterone in serum can provide information to evaluate adrenal and testicu-lar functions. It is useful for the diagnosis of the hypergonadism (1) and hyperpituita-rism(2) in man, and hirsutism(3), menstrual disorders(4), and polycystic ovarian syn-drome(5) in woman. It is also useful for the characterization and follow-up of some cancers such as testes(6), breast, ovarian, and adrenal tumours(7-9).

Principle of the Assay The ST AIA-PACK Testosterone is a competitive immunoenzymometric as-say, which is performed entirely in the AIA-PACK. Testosterone present in the test sample competes with enzyme-labeled testosterone for a limited num-ber of binding sites on the testosterone specific monoclonal antibody immobi-lized on a magnetic solid phase. The magnetic beads are washed to remove unbound enzyme-labeled testosterone and are then incubated with a fluoro-genic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of en-zyme-labeled testosterone that binds to the beads is inversely proportional to the testosterone concentration in the test sample. A standard curve is constructed, and unknown sample concentrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume 70 µl Specimen type: Serum Assay range: up to 2200 ng/dl Calibration stability: 90 days Sensitivity: 7 ng/dl

Reference range

Male 262 – 870 ng/dl Female 9.0 – 56.0 ng/dl

TTeessttoosstteerroonnee

SPECIFICITY The following substances were tested for cross-reactivity. Cross-reactivity is ex-pressed in term of the percentage of the concentration of each substance which will be measured as testosterone. Substance Concen-

tration added (ng/dL)

% cross reactivity (mole %)

Androstenedione 200000 1.71 Aldosterone 160000 0.00 Androsterone 2000000 0.03 Corticosterone 100000 0.01 Cortisol 160000 0.00 Cortisone 160000 0.00 Danazol 4000 0.00 11-Deoxycortisol 20000 0.00 Dexamethasone 160000 0.00 DHEA 5000 0.00 DHEA-sulfate 20000 0.00 5a-Dihydrotestosterone 1000 0.81 Esteadiol 200000 0.00 Estrone 10000 0.00 Ethisterone 1000 0.00 Fluoxymesterone 100000 0.00 19-Hydroxy-androstenedione

5000 0.04

Methyltestosterone 600 0.14 Norethindrone 500 0.00 Prednisone 16000 0.00 Progesterone 20000 0.02 Norethynodrel 1000 0.03 Spironolactone 20000 0.01 Triamcinolone 1000 0.00

Ver 05-01

PRECISION The intra-assay (within run) precision coefficient of variation was evaluated in three serum and samples by 10 replicate determinations in five different lots of reagent.

Rgt. Lot

Sample


Mean (ng/dL)

Standard Deviation (ng/dL)

Coefficient of Variation (%)

BZ12472

Serum A Serum B Serum C

10 10 10

52.0 800 1633

1.80 19.6 45.9

3.5 2.4 2.8

BZ12473


10 10 10

53.0 798 1558

2.79 15.4 28.2

5.3 1.9 1.8

C212474


10 10 10

61.4 796 1503

3.19 25.9 46.0

5.2 3.3 3.1

The inter-assay (between run) precision coefficient of variation was evaluated at three different concentrations by analyzing control samples in 20 separate runs.

Sample


Mean (ng/dL)

Standard Deviation (ng/dL)

Coefficient of Variation (%)

Sample A Sample B Sample C

20 20 20

88.6 627.2 1402

5.31 16.8 34.8

5.99 2.69 2.48

SENSITIVITY The minimal detectable concentration (MDC) of testosterone is estimated to be 7 ng/dL . The MDC is defined as that concentration of testosterone, which corresponds to the rate of fluores-cence that is two standard deviations from the mean rate of fluorescence of 20 replicate deter-minations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

Added hemoglobin (up to 411.8 mg/dL), conjugated bilirubin (up to 18.2 mg/dL) and free bilirubin (up to 17 mg/dL) do not interfere with the assay.

Lipemia, as indicated by added triglyceride (up to 500 mg/dL) does not interfere with the assay.

Added albumin (up to 0.5 g/dL) does not interfere with the assay. Added ascorbic acid (up to 20 mg/dL) does not interfere with the assay. Added rheumatoid factor (up to 45 IU/ml) does not interfere with the assay. Heparin (up to 100 U/ml) does not interfere with the assay

ST AIA-PACK Test 025204 Calibrator Set 020304 Sample Diluting Solution 020504 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Anti-thyroid antibodies, especially anti-microsomal antibodies and anti-thyroglobulin antibodies (TgAb) have been shown to be relevant to autoim-mune thyroid disease such as Graves’ disease and Hashimoto’s dis-ease(1).However, TgAb sometimes test positive for autoimmune diseases other than those involving the thyroid, so the disease specificity of TgAb is not clear (2,3). Thyroglobulin (Tg) is a protein that has a molecular weight of about 660 kDa. It is synthesised in thyroid gland cells and is the main component of the colloid con-tained in the thyroid follicles. Tg plays an important role in the synthesis and pres-ervation of thyroid hormones. Although TgAb have conventionally been assayed by semi-quantitative methods using indi-rect agglutination, highly sensitive quan-titative methods such as the RIA and EIA methods have been developed (4–9). AIA-PACK TgAb is an enzyme immuno-assay reagent to measure concentra-tions of TgAb in human serum using Tg antigen coated beads and anti-human IgG antibody labelled with alkaline phosphatase.

PRINCIPLE OF THE ASSAY The AIA-PACK TgAb is a two-step im-munoenzymometric assay, which is per-formed in the AIA cup. TgAb present in test sample are bound to Tg antigen immobilised on the beads. The beads are washed to remove unbound materi-als and are then incubated with enzyme-labelled monoclonal anti-human IgG an-tibody. The beads are washed again to remove unbound enzyme-labelled monoclonal antibody and are then incu-bated with a fluorogenic substrate. The amount of enzyme-labelled antibody that binds to the beads is directly propor-tional to the TgAb concentration in the test sample. A standard curve using a range of known standard concentrations is constructed and unknown sample TgAb concentrations are calculated us-ing this curve.

Incubation time: 40 minutes Specimen volume 50 µl Specimen type: Serum Assay range: up to 40 IU/ml Calibration stability: 90 days Sensitivity: 0.12 IU/ml

Reference range

<13.6 IU/ml

TTgg AAbb

Ver 05-01

PRECISION The intra-assay coefficient of variation was evaluated in three samples by 10 replicate determinations.

Sample Nr of Rep-licates

Mean (IU/ml)

Standard De-viation (IU/ml)


(%) Sample L 10 2.91 0.134 4.6 Sample M 10 11.92 0.489 4.1 Sample H 10 22.59 0.700 3.1

The inter-assay coefficient of variation was evaluated at two different concentrations by analysing samples in 20 separate runs.

Sample Nr of Rep-licates

Mean (IU/ml)

Standard Deviation

(IU/ml)


(%) Sample L 20 2.96 0.084 2.8 Sample H 20 23.54 0.592 2.5

SENSITIVITY The minimal detectable concentration of human TgAb using the AIA-PACK TgAb kit is estimated to be 0.12 IU/ml, which corresponds to 6 IU/ml in original serum sample (dilu-tion factor 51). The minimal detectable concentration is defined as that concentration of human TgAb which corresponding to the rate of fluorescence that is two Standard De-viations from the mean rate of fluorescence of 20 replicate determinations of Calibrator 1. INTERFERENCES Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

Hemoglobin (up to 454 mg/dl), free bilirubin (up to 13.8 mg/dl) and conjugate bilirubin (up to 19.9 mg/dl) do not interfere with the assay.

Lipemia, as indicated by triglyceride concentrations (up to 293 mg/dl), does not interfere with the assay.

Protein, as indicated by albumin concentrations (up to 5 g/dl), does not interfere with the assay.

Ascorbic acid (up to 20 mg/dl) does not interfere with the assay.

ST AIA-PACK TgAb 025291 Calibrator Set 020391 Sample Diluting Solution 020591 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Anti-thyroid antibodies, especially anti-microsomal antibodies and anti-thyroglobulin antibodies have been shown to be relevant to autoimmune thy-roid disease such as Graves' disease and Hashimoto's disease. For the detec-tion of the anti-microsomal antibodies, some methods based on the thyroid mi-crosomal fraction coated cell or particles were available. Recently, the major component of the antigen recognised by the anti-microsomal antibodies was identified as the thyroid peroxidase (TPO Ab)(1-2) The findings have led to some sensitive and specific assays for the anti-microsomal antibodies that are anti-TPO antibodies (TPO Ab) using TPO antigen (3-8). AIA-PACK TPO Ab is an enzyme immu-noassay reagent to measure concentra-tions of TPO Ab in human serum using TPO antigen coated beads and anti-human IgG antibody labelled with alka-line phosphatase.

PRINCIPLE OF THE ASSAY The AIA-PACK TPO Ab is a two-step immunoenzymometric assay, which is performed in the AIA cup. TPO Ab pre-sent in test sample is bound to TPO an-tigen immobilised on the beads. The beads are washed to remove unbound materials and are then incubated with enzyme-labelled monoclonal anti-human IgG antibody. The beads are washed again to remove unbound enzyme-labelled monoclonal antibody and are then incubated with a fluorogenic sub-strate. The amount of enzyme labelled antibody that binds to the beads is di-rectly proportional to the TPO Ab con-centration in the test sample. A standard curve using a range of known standard concentrations is constructed and un-known sample TPO Ab concentrations are calculated using this curve.

Incubation time: 40 minutes Specimen volume 10 µl Specimen type: Serum Assay range: up to 20 IU/ml Calibration stability: 90 days Sensitivity: 0.05 IU/ml

Reference range

<3.2 IU/ml

TTPPOO AAbb

Ver 05-01

PRECISION The intra-assay coefficient of variation was evaluated in three samples by 10 replicate determinations Sample Number of Mean Standard Coefficient Replicates Deviation of Variation (IU/ml) (IU/ml) (%) Sample L. 10 1.91 0.068 3.6 Sample M 10 10.30 0.333 3.2 Sample H 10 17.10 0.753 4.4 The inter-assay coefficient of variation was evaluated at two different concentrations by analysing samples in 20 separate runs. Sample Number of Mean Standard Coefficient Replicates Deviation of Variation (IU/ml) (IU/ml) (%) Sample L 20 1.90 0.049 2.6 Sample H 20 15.82 0.402 2.5 SENSITIVITY The minimal delectable concentration of human TPO Ab using the AIA-PACK TPO Ab kit is estimated to be 0.05 IU/ml, which corresponds to 2.5 IU/ml in original serum sam-ple (dilution factor 51). The minimal detectable concentration is defined as that concen-tration of human TPO Ab, which corresponds to the rate of fluorescence that is two Standard Deviations from the mean rate of fluorescence of 20 replicate determinations of Calibrator 1. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

Hemoglobin (up to 411 mg/dl,), free bilirubin (up to 13.8 mg/dl) and conjugate bilirubin (up to 19.9 mg/dl) do not interfere with the assay.

Lipemia, as indicated by triglyceride concentrations (up to 293 mg/dl), does not interfere with the assay.

Protein, as indicated by albumin concentrations (up to 5 g/dl) does not interfere with the assay.

Ascorbic acid (up to 20 ml/dl) does not interfere with the assay. The results were obtained by adding these interfering substances to diluted samples.

ST AIA-PACK TPO 025290 Calibrator Set 020390 Sample Diluting Solution 020590 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Troponin exists as a complex of three distinct proteins: troponin T, troponin C, and troponin I1). The troponins regulate the calcium-modulated interaction of actin and myosin in striated (skeletal and cardiac) muscle with tro-ponin I acting as the actinomyosin ATPase inhibitor. Three isoforms of troponin I have been identi-fied2) which are products of different genes with unique amino acid sequences3). Cardiac troponin I (cTnI) is found exclusively in cardiac tissue 4). Though the three forms of troponin I share some homologous structure, cTnI has a sequence of 31 amino acid residues at the amino terminus of the molecule which renders cTnI distinctly antigenically different from the other isoforms of troponin I, as well as tro-ponin C and troponin T 5). Monoclonal antibod-ies developed to react with epitopes in this unique portion of the cTnI molecule have made possible the development of in vitro di-agnostic assays with great specificity for cTnI. Biochemical markers of myocardial injury are considered the "gold standard" for the diagno-sis of AMI and are particularly important in the nondiagnostic EKG patients 6, 7). Clinical inves-tigations have shown elevated cTnI concentra-tions to be extraordinarily useful in aiding the diagnosis of cardiac specific injury in patients with acute myocardial infarction 8), myocardial contusion 9), and unstable angina 10). Following an AMI, cTnI is released into the bloodstream from the damaged heart muscle and within 4 - 8 hours, levels of cTnI increase above levels established for non-AMI specimens. Peak concentrations are generally reached 12 - 18 hours after the AMI onset and cTnI concentra-tions remain elevated for 5 - 10 days11, 12, 13). Troponin I shows a temporal rise and fall in AMI patients similar to that seen with CKMB mass. However, recent studies indicate that because of its cardiac specificity, cTnI is much more useful than CKMB in detecting myocar-dial injury especially in the presence of skele-tal muscle damage and lesions (rhabdomyoly-sis, polytraumatism)3, 10,14,15).

PRINCIPLE OF THE ASSAY The AIA-PACK cTnI 2nd-Gen is a two-site immunoenzymometric assay which is performed entirely in the AIA-PACK. Troponin I present in the test sample is bound with monoclonal antibody immo-bilized on a magnetic solid phase and enzyme-labeled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labeled monoclonal antibody that binds to the beads is directly proportional to the tro-ponin I concentration in the test sample. A standard curve is constructed, and unknown sample concentrations are cal-culated using this curve. AIA-PACK cTnI 2nd-Gen replaces the original AIA-PACK cTnI product and has been labeled as second generation to avoid any possible confusion.


Reference range

With clinical specificity equal to 100% using <0.1 ng/mL as a range of values for AMI cut-off.

TTrrooppoonniinn II

Ver 05-01

PRECISION The intra-assay (within run) precision coefficient of variation was evaluated in three serum sam-ples and two plasma samples by 10 replicate determinations in five different lots of reagent on the AIA-21. Sample Number of Mean Deviation CV Replicates (ng/mL) (ng/mL) (%) Serum A 10 3.48 0.12 3.3 Serum B 10 8.05 0.24 3.0 Serum C 10 43.90 0.54 1.2 Plasma A 10 4.63 0.07 1.6 Plasma B 10 26.54 0.58 2.2 The inter-assay (between run) precision coefficient of variation was evaluated at five different concentrations by analyzing control samples in 20 separate runs on the AIA-21. Sample Number of Mean Deviation Coefficient of Replicates (ng/mL) (ng/mL) Variation (%) Serum A 20 4.67 0.09 1.9 Serum B 20 10.48 0.26 2.5 Serum C 20 50.76 1.18 2.3 Plasma A 20 6.42 0.18 2.9 Plasma B 20 32.48 0.67 2.1 SENSITIVITY The minimal detectable concentration (MDC) of troponin I is estimated to be 0.02 ng/mL. The MDC is defined as that concentration of troponin I which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. These studies were run on the AIA-600 II on multiple lots of reagents INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration. • Added hemoglobin (up to 436 mg/dL), conjugated bilirubin (up to 18.6 mg/dL) and

free bilirubin (up to 16.5 mg/dL) do not interfere with the assay. • Lipemia, as indicated by added triglyceride (up to 2000 mg/dL) does not interfere with

the assay. • Added albumin (up to 2.5 g/dL) does not interfere with the assay. • Added ascorbic acid (up to 20 mg/dL) does not interfere with the assay. • Added rheumatoid factor (up to 45 IU/ml) does not interfere with the assay. • Heparin (up to 50 U/ml) does not interfere with the assay.

ST AIA-PACK Troponin 025205 Calibrator Set 020305 Sample Diluting Solution 020505 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Thyroid Stimulating Hormone is a glyco-protein hormone secreted by the anterior pituitary gland. When feedback suppres-sion of the pituitary is reduced by a re-duced production of thyroid hormones (T4 and T3), TSH rises in an attempt to in-crease thyroid hormone production. This rise occurs while the patient is still asymp-tomatic and thus is an early and very sen-sitive indication of hypothyroidism(1-5). TSH is also controlled by the hypothalamic pep-tide, thyrotropin releasing hormone (TRH).Accurate determination of serum TSH is the most useful and sensitive test for pri-mary hypothyroidism, where serum thyroid hormone concentrations are depressed and serum TSH concentrations are signifi-cantly elevated. Serum TSH determina-tions may also be used to differentiate be-tween pituitary (secondary) and hypotha-lamic (tertiary) hypothyroidisms (6-9). Through the use of monoclonal antibody technology, which provides the necessary specificity and sensitivity, the usefulness of TSH determination in the diagnosis of hyperthyroidism distinguished from euthy-roidism has been well established.

PRINCIPLE OF THE ASSAY The ST AIA-PACK TSH is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK test cups. TSH present in the test sample is bound with monoclonal antibody immo-bilized on magnetic beads and mono-clonal antibody conjugated with bovine alkaline phosphatase in the AIA- PACK test cups. The magnetic beads are washed to remove unbound enzyme-labeled monoclonal antibody and are then incubated with a fluorogenic sub-strate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme conju-gated with monoclonal antibody that binds to the beads is directly propor-tional to the TSH concentration in the test sample. A standard curve is con-structed, and unknown sample concen-trations are calculated using this curve

Incubation time: 10 minutes Specimen volume 100 µl Specimen type: Serum Assay range: up to 100 µIU/ml Calibration stability: 90 days Sensitivity: 0.01 µIU/ml

Reference range

0.39 – 3.55 µIU/ml

TTSSHH

SPECIFICITY The following substances were tested for cross-reactivity. Cross-reactivity is expressed in terms of the percentage of the concentra-tion of each substance, which will be meas-ured as TSH.

Compound Cross-reactivity(%) TSH 100 HCG < 0.01 FSH < 0,01 LH 0.17 HGH < 0.01

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calculate the coeffi-cient of variation (CV). Sample Mean Pooled SD CV ( µIU/ml) ( µIU/ml) (%) Serum A3 2.19 0.064 2.9 Serum B3 8.26 0.192 2.3 Serum C3 80.63 2.016 2.5 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and co-efficient of variation (CV). Sample Mean Pooled SD CV ( µIU/ml) ( µIU/ml) (%) Serum A3 2.19 0.110 5.0 Serum B3 8.26 0.399 4.8 Serum C3 80.63 3.577 4.4 SENSITIVITY The minimal detectable concentration (MDC) of TSH is estimated to be 0.01 µIU/ml. The MDC is defined as that concentration of TSH, which corresponds to the rate of fluores-cence that is two standard deviations from the mean rate of fluorescence of 30 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 390 mg/dL), free bilirubin (up to 17,1 mg/dL) and conjugated bilirubin (up to 18.2 mg/dL) do not interfere with the assay.


- Protein, as indicated by human serum albumin concentrations (up to 50 mg/ml). does not interfere with the assay.


ST AIA-PACK TSH 025294 Calibrator Set 020394 Sample Diluting Solution 020594 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Triiodothyronine (T3) and thyroid hor-mone, (thyroxine;T4), regulate a variety of biochemical processes throughout the body (1). The majority of T3 in circulation is produced enzymatically by monodeio-dination of T4 in the peripheral tissues, rather than from direct secretion from the thyroid gland (2). Approximately one-third of all T4 secreted is deiodinated to yield T3 (3). Serum T3 measurements can be a valu-able component of a thyroid-function screening panel in diagnosing certain disorders of thyroid function in addition to conditions caused by iodide defi-ciency. Assays for T3 are valuable in early detection of hyperthyroidism and for monitoring the efficacy of treatment for thyroid disorders (4). A normal T3 value in the presence of an elevated T4 and/or Free T4 (FT4) level may also help to rule out hyperthyroidism (5).

PRINCIPLE OF THE ASSAY The ST AIA-PACK TT3 is a competitive enzyme immunoassay, which is per-formed entirely within the AIA-PACK. Triiodothyronine, which is displaced from its binding proteins by ANS (8-anilino-1-naphthalene sulfonic acid), and free T3 present in the test sample compete with enzyme-labeled T3 for a limited number of binding sites on a T3 specific antibody immobilized on magnetic beads. The beads are washed to remove the un-bound enzyme-labeled T3 and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4-MUP). The amount of enzyme labeled T3 that binds to the beads is inversely proportional to the T3 concentration in the test sample. A standard curve using a range of known standard concentra-tions is prepared and unknown T3 con-centrations are calculated using this curve.

Incubation time: 10 minutes Specimen volume 25 µl Specimen type: Serum Assay range: up to 8.0 ng/ml Calibration stability: 90 days Sensitivity: 0.25 ng/ml

Reference range

0.79 - 1.58 ng/mL

TTTT33

SPECIFICITY The following substances were tested for cross-reactivity. Cross-reactivity is expressed in terms of the percentage of the concentration of each substance which will be measured as T3. Compound Cross-reactivity L-Triiodothyronine (L-T3) 100 % D-Triiodothyronine (D-T3) 125 L-Thyroxine (L-T4) 0.15 D-Thyroxine (D-T4) 0.07 3, 3’, 5’-Triiodothyronine 0.3 3, 5-Diiodo-L-tyrosine < 0.001

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV (ng/mL) (ng/mL) (%) Serum A3 0.71 0.027 3.8 Serum B3 1.72 0.040 2.3 Serum C3 3.72 0.090 2.4 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV (ng/mL) (ng/mL) (%) Serum A3 0.71 0.044 6.3 Serum B3 1.72 0.063 3.7 Serum C3 3.72 0.119 3.2 SENSITIVITY The minimal detectable concentration (MDC) of T3 is estimated to be 0.2 ng/mL. The MDC is defined as that concentration of T3, which corresponds to the rate of fluores-cence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.



- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay.

ST AIA-PACK TT3 025282 Calibrator Set 020382 Sample Diluting Solution 020582 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Evaluation of thyroid status is complex. The primary function of the thyroid gland is the secretion of thyroxine (T4) or triio-dothyronine (T3). Abnormal secretion of T4 and/or T3 may lead either to hyper- or hypo-thyroidism. The synthesis and release of T4 and T3 are in response to a hypothalamic-pituitary signal, Thyroid Stimulating Hormone (TSH), which is released from the anterior pituitary and is the principal regulator of thyroid activ-ity (1). The release of TSH is controlled by Thyrotropin Releasing Hormone (TRH) from the hypothalamus(2). This combined system regulating the release of thyroid hormone is the hypothalamic-pituitary axis (3,4). In the circulation, T4 is 99.97% protein bound (0.03% free) while T3 is 99.7% bound (0.3% free). Thyroxine Binding Globulin (TBG) is the primary binding protein. To a lesser ex-tent, Thyroxine Binding Prealbumin (TBPA) and albumin can also bind T4 (5,6,7). Only unbound (free) forms exert the physiological actions. T4 is largely converted to T3 in periph-eral tissues by monodeiodinatio (8). Total T4 rises and falls with the TBG level in euthyroid individuals. An erroneous in-terpretation of thyroid function may be obtained if a condition exists which changes the TBG concentrations. Cer-tain drugs compete with T4 for binding to TBG, which results in decreased levels of total T4 through the negative feed-back of thyroid hormone concentration on TSH secretion (9,10).

PRINCIPLE OF THE ASSAY The ST AIA-PACK T4 is a competitive enzyme immunoassay, which is per-formed entirely within the AIA-PACK. Thyroxine, which is displaced from its binding proteins by ANS (8-anilino-1-naphthalene sulfonic acid), and free T4 present in the test sample compete with enzyme-labeled thyroxine for a limited number of binding sites on a thyroxine-specific antibody immobilized on mag-netic beads. The beads are washed to remove the unbound enzyme labeled thyroxine and are then incubated with a fluorogenic substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme labeled thyroxine that binds to the beads is inversely pro-portional to the thyroxine concentration in the test sample. A standard curve us-ing a range of known standard concen-trations is constructed and unknown thy-roxine concentrations are calculated us-ing this curve.

Incubation time: 10 minutes Specimen volume 10 µl Specimen type: Serum Assay range: up to 24 µg/dl Calibration stability: 90 days Sensitivity: 0.5 µg/dl

Reference range

4.0 - 11.0 µg/dL

TT44

Ver 05-01

PRECISION Within run precision was determined using three controls in a total of 20 runs. Within each run, one set of duplicates per control was assayed. The mean of each duplicate was used to obtain the pooled standard deviation (SD), which was then used to calcu-late the coefficient of variation (CV). Sample Mean Pooled SD CV ( µg/dL) ( µg/dL) (%) Serum A3 2.23 0.135 6.1 Serum B3 6.86 0.266 3.9 Serum C3 15.14 0.460 3.0 Total precision was determined by the duplicate assay of three controls in 20 separate runs. The means of each run were used to calculate the standard deviation (SD) and coefficient of variation (CV). Sample Mean SD CV ( µg/dL) ( µg/dL) (%) Serum A3 2.23 0.138 6.2 Serum B3 6.86 0.295 4.3 Serum C3 15.14 0.601 4.0 SENSITIVITY The minimal detectable concentration (MDC) of thyroxine is estimated to be 0.5 µg/dL. The MDC is defined as that concentration of thyroxine, which corresponds to the rate of fluorescence that is two standard deviations from the mean rate of fluorescence of 20 replicate determinations of a zero calibrator. INTERFERENCE Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.



- Ascorbic acid (up to 20 mg/dL) does not interfere with the assay.

ST AIA-PACK T4 025258 Calibrator Set 020358 Sample Diluting Solution 020558 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

SUMMARY AND EXPLANATION OF TEST Prostate Specific Antigen (PSA) or Pros-tate Antigen (PA) was identified in 1977 by Wang et al (1) PA is a single chain glycoprotein of approximately 34 kDal-tons containing 7% carbohydrate(2). The presence of PSA has been demon-strated in normal, benign hyperplastic and malignant prostatic tissue, in metas-tatic prostatic carcinoma, and also in prostatic fluid as well as seminal plasma(5). PSA is not present, however, in any other tissue from men, nor is it produced by cancers of the breast, lung, colon, rectum, stomach, pancreas or thyroid(6) Elevated serum PA levels have been reported in patients with prostate cancer, benign prostatic hypertrophy, or inflammatory conditions of other adja-cent genitourinary tissues.(3,7)

In serum it exists in the free form and complexed to a series of protease inhibi-tors. PSA in serum forms stable com-plexes with ACT and α2-macroglobulin. The ACT-PSA complex represents a predominant fraction of circulating PSA, however less for patients with Benign Prostate Hyperplasia (BPH) than those with prostate cancer. ACT-PSA complex and free fraction PSA are both immuno detectable forms of PSA in serum. Uncomplexed fraction PSA assists in discriminating between BPH and prostatic cancer, in addition to total PSA. It has been shown that the determination of the different forms of PSA is helpful to discriminate between prostatic carci-noma and Hyperplasia. Among patients with elevated PSA concentrations, the men with prostatic carcinoma have a tendency to have relatively more ACT-PSA complex present than the men hav-ing BPH.

PRINCIPLE OF THE ASSAY The ACT-PSA complex is precipitated, after which the uncomplexed PA fraction of the sample is determined with the AIA-PACK ucPA. The AIA-PACK ucPA is a two-site immunoenzymometric as-say, which is performed entirely in the AIA-PACK. Uncomplexed fraction PSA present in the pre-treated test sample is bound with monoclonal antibody immo-bilised on magnetic beads and enzyme-labelled monoclonal antibody in the AIA-PACK. The magnetic beads are washed to remove unbound enzyme-labelled monoclonal antibody and are then incu-bated with a fluorogenic Substrate, 4-methylumbelliferyl phosphate (4MUP). The amount of enzyme-labelled mono-clonal antibody that binds to the beads is directly proportional to the uncomplexed fraction PSA concentration in the test sample.


Reference range

See pack insert

uuccPPAA

Ver 05-01

PRECISION The intra-assay coefficient of variation was evaluated in two control samples by 20 rep-licate determinations in three different runs. Sample Number of Mean Standard Coefficient of

Replicates (ng/mL) Deviation (ng/mL) Variation (%) Serum A 20 1.52 0.09 6.1 Serum B 20 8.49 0.47 5.5 Serum C 20 27.92 0.83 3.0 The inter-assay coefficient of variation was evaluated in two control samples by 20 rep-licate determinations. Sample Number of Mean Standard Coefficient of

Replicates (ng/mL) Deviation (ng/mL) Variation (%) Serum A 20 1.40 0.08 5.8 Serum B 20 8.01 0.33 4.1 Serum C 20 25.79 1.22 4.7 SPECIFICITY The following substance was tested for cross-reactivity. Cross reactivity is expressed in terms of the percentage of the concentration of each substance. Compound Cross-reactivity (%) PAP 0.0 SENSITIVITY The minimal detectable concentration (MDC) of ucPA is estimated to be 0.01 ng/mL. The MDC is defined as that concentration of free fraction PSA that corresponds to the rate of fluorescence that is two Standard Deviations from the estimated mean rate of fluorescence of 20 replicates of Calibrator 1. INTERFERENCES Interference is defined, for purposes of this study, to be recovery outside of 10% of the known specimen mean concentration.

- Hemoglobin (up to 470 mg/dL) and free bilirubin (up to 17 mg/dL) do not interfere with the assay.

- Lipemia, as indicated by triglyceride concentrations (up to 1.600 mg/dL), does not interfere with the assay.

- Added Protein (up to 50 mg/dL) does not interfere with the assay.

ST AIA-PACK ucPA 025202 Calibrator Set 020302 Sample Diluting Solution 020502 Substrate Set II 020968 Wash Concentrate 020955 Diluent Concentrate 020956

analyte sheets-english - pribori.compribori.com/products/pdf/tosoh_reagent.pdf · the aia-pack b12...

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