dbc’s new aldosterone elisa kit includes a ......1. primary hyperaldosteronism causes salt and...
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384 Neptune CrescentLondon, Ontario, Canada N6M 1A1
Tel: 519-681-8731Fax: [email protected]
Manufacturing Innovative IVDfor the World Since 1973
DBC’s NEW ALDOSTERONE ELISA KIT INCLUDES A READY-TO-USE CONJUGATE
AND BLOCKING AGENTS
REF CAN-ALD-500
significantproportionofhypertensiveindividualssufferfromresistanthyper-
tensionduetofactorssuchasnon-compliance,poornutritionalpractice(highsalt,alcohol,licorice)orsecondaryhypertension(20%ofallresistanthypertensioncases).Diagnosingthecauseofhypertensionisofparamountimportancetoselectthecorrecttherapy.
Heartfailure,stroke,renalconditionsanddementiaaresomeofthecommon
consequencesofuncontrolledhyper-tension;theoccurrenceoftheseconditionshowever,canbereducedwhentheunderlyingsourceofresistanthypertensionisidentifiedandafollowuptherapyisapplied.
Renin
Angiotensinogen
Aldosterone
Angiotensin I
Angiotensin II
ACE
THE RENIN-ANGIOTENSIN-ALDOSTERONE AXISplaysakeyroleinresistanthypertension.
Duringnormalhomeostasis,reninisreleasedunderconditionsofdehydrationorlowbloodpressure(seefigurebelow).Renin enzymatic activitythenpromotesthecleavageofAngiotensinogenandgenerationofAngiotensinI,whichinturnistransformedintoAngiotensinIIandactivatesaldosteronerelease(whichcausessaltandwaterretention,andexcretionofpotassium,magnesium,andotherions).
A 1. SpenceJD.LessonsfromAfrica:theimportanceofmeasuringplasmareninandaldosteroneinresistanthypertension.Can J Cardiol.2012;28(3):254–7.
2. SpenceJD.Physiologictailoringoftherapyforresistanthypertension:20years’experiencewithstimulatedreninprofiling.Am J Hypertens.1999;12:1077–83.
3. SpenceJD.Physiologictailoringoftreatmentinresistanthypertension.Current Cardiology Reviews.2010;6:119–123.
4. JonesES,etal.PhysiologicallyIndividualizedTherapyforResistantHypertensioninAfrica.HypertensionTeachingSeminarOrganizedbytheInternationalSocietyofHypertension(ISH)AfricaRegionalAdvisoryGroupincollaborationwiththeEuropeanSocietyofHypertension(ESH),theInternationalForumforHypertensionControlandPreventioninAfrica(IFHA)andtheMozambicanHeartAssociation(AMOCOR);April18–19,2016;Maputo,Mozambique2016.
LITERATURE
new
at a glanceCatalogue number: CAN-ALD-500Number of test wells: 96Sensitivity: 9.1 pg/mLSample Volume: 50 µLTotal assay time: 80 mins.Validated against: LC-MS/MS
1. Primaryhyperaldosteronismcausessaltandwaterretention,feedingbacktosuppressreninactivity.
2. Renalorrenovascularcausesofhypertensionleadtoelevatedreninactivitywithsecondaryhyperaldosteronism.
3. Impairmentoftherenaltubularepithelialsodiumchannel(suchasLiddle’ssyndrome)causessaltandwaterretentionandsuppressesbothreninactivityandaldosterone.
ALDOSTERONE MEASUREMENTisthereforeanoutstandingtooltodeterminethephysiologicalcausesofresistanthypertension,permittingthephysiciantochoosethemostappropriatetherapy.1-3
Thefollowingalgorithmwasusedinastudyofresistanthypertensioninthreehyper-tensionclinicsinAfrica,inastudyfundedbyGrandChallengesCanada.Thisapproachincreasedsystolicbloodpressurecontrolfrom25%inusualcareto75%inindividualizedcarebasedonaldosterone/reninprofiling.4
PrimaryAldosteronism Liddle’sVariants,AdducinPolymorphisms
RenalorRenovascular
Aldosterone High Low High
Renin Low Low High
Primarytreatment Aldosteroneantagonists:SpironolactoneEplerenone(Amilorideformenwhereeplerenoneisnotavailable)(rarelysurgery)
AmilorideAngiotensinreceptorblockersAliskiren(rarelyrevascularization)
Thepreciseandaccuratemeasurementofaldosteronebyenzymeimmunoassaycanbeanimportanttoolforthediagnosisoftheunderlyingcauseofhypertension,leadingtoappropriatetherapy.
Thisapproachnotonlyimprovesbloodpressurecontrol,thusreducingtheriskofstroke,heartfailureandrenalfailure,butalsoreducesadverseeffectsofmedicationandmayreducethecostofmedicationbyidentifyingspecifictherapy.
When this metabolism is altered, three patterns of aldosterone and renin activity levels can be produced:
PERFORMANCE
TheRfBinternationalcontrolswereassayedwiththeDBCELISAkit(CAN-ALD-500).Theresultsforthekitarethemean±SDof4independentexperiments(pg/mL).
TheresultsmatchbothLC-MS/MSresultsfromMayoClinicandfallwithintherangeofresultsestablishedfromallmethods.
RfB lot HM 2/15 A HM 2/15 BLC-MS/MS(MayoClinic) 570 110
LC-MS/MS(RfB)
Target
634 122
Range (16P–84P)
419–764 111–172
AllMethods(RfB)
Target
529 110
Range (16P–84P)
468–658 90–144
CAN-ALD-500(DBC) 636±70 105±16
Evaluation of International Controls
Aldosteroneconcentrationrangeinthepopulationdependsontheethnicandsocialcompositionandnutritionalfactors.
Each laboratory must determine its own reference ranges!
Aldosterone (pg/mL)
Freq
uenc
y
120 Putatively healthy individuals—UprightMedian:52pg/mL
Average:70pg/mL
95%confidencerange:ND–210pg/mL
REFERENCE RANGE
DBC has launched a new Aldosterone kit(CAN-ALD-500) that includes a ready-to-use conjugate and blocking agents that prevent interferences by sample endogenous substances.
PROCEDURE
ASSAY PRINCIPLEThenewDBCAldosteroneELISAkit(CAN-ALD-500)isacompetitiveimmunoassaythatusesinnovativechemistryandaspecificanti-aldosteroneantibodythatbindsquantitativelytoallisomersofaldosterone.
50μLcalibrators/samples
100μLofReady-to-UseConjugate
1hroomtemperature/shaking
Wash3x
150μLTMB
20minroomtemperature/shaking
50μLofstopsolution
Readinaplatereaderat450nm
Aldosterone (pg/mL)
OD
(450
nm
)
Typical Calibration CurveSamplecurveonly.
Donotusetocalculateresults.
Parameter DBC Competitor 1 Competitor 2 LC-MS/MSTotalassaytime 1h20min 1h30min Overnight+1h N/A
Readytousereagents Yes Yes No N/A
DynamicRange,pg/mL 10–1000 5.7–1000 4.7–250 40–1000
Samplesize,μL 50 50 100 600
Samplepre-treatment
Serum,Plasma No No Yes Yes
Urine No Yes Yes Yes
Sensitivity,pg/mL 9.1 5.7 4.7 40
Precision,CV%
Intra-assay 5.5–9.4 3.8–9.7 4.5–6.6
Inter-assay 7.6–12.8 8.6–11.5 10.8–16.3
ComparativeanalysisofserumsamplesresultsbetweenthenewDBCAldosteronekit(CAN-ALD-500)andLC-MS/MSperformedatMayoClinic.
LC-MS/MS Aldosterone (pg/mL)
DB
C A
ldos
tero
ne E
LISA
(pg/
mL) y=1.0578x+52.782
r=0.95,n=36
PERFORMANCE
DBC has launched a new Aldosterone kit(CAN-ALD-500) that includes a ready-to-use conjugate and blocking agents that prevent interferences by sample endogenous substances.
PROCEDURE
ASSAY PRINCIPLEThenewDBCAldosteroneELISAkit(CAN-ALD-500)isacompetitiveimmunoassaythatusesinnovativechemistryandaspecificanti-aldosteroneantibodythatbindsquantitativelytoallisomersofaldosterone.
50μLcalibrators/samples
100μLofReady-to-UseConjugate
1hroomtemperature/shaking
Wash3x
150μLTMB
20minroomtemperature/shaking
50μLofstopsolution
Readinaplatereaderat450nm
Aldosterone (pg/mL)
OD
(450
nm
)
Typical Calibration CurveSamplecurveonly.
Donotusetocalculateresults.
Parameter DBC Competitor 1 Competitor 2 LC-MS/MSTotalassaytime 1h20min 1h30min Overnight+1h N/A
Readytousereagents Yes Yes No N/A
DynamicRange,pg/mL 10–1000 5.7–1000 4.7–250 40–1000
Samplesize,μL 50 50 100 600
Samplepre-treatment
Serum,Plasma No No Yes Yes
Urine No Yes Yes Yes
Sensitivity,pg/mL 9.1 5.7 4.7 40
Precision,CV%
Intra-assay 5.5–9.4 3.8–9.7 4.5–6.6
Inter-assay 7.6–12.8 8.6–11.5 10.8–16.3
ComparativeanalysisofserumsamplesresultsbetweenthenewDBCAldosteronekit(CAN-ALD-500)andLC-MS/MSperformedatMayoClinic.
LC-MS/MS Aldosterone (pg/mL)
DB
C A
ldos
tero
ne E
LISA
(pg/
mL) y=1.0578x+52.782r=0.95,n=36
PERFORMANCE
1. Primaryhyperaldosteronismcausessaltandwaterretention,feedingbacktosuppressreninactivity.
2. Renalorrenovascularcausesofhypertensionleadtoelevatedreninactivitywithsecondaryhyperaldosteronism.
3. Impairmentoftherenaltubularepithelialsodiumchannel(suchasLiddle’ssyndrome)causessaltandwaterretentionandsuppressesbothreninactivityandaldosterone.
ALDOSTERONE MEASUREMENTisthereforeanoutstandingtooltodeterminethephysiologicalcausesofresistanthypertension,permittingthephysiciantochoosethemostappropriatetherapy.1-3
Thefollowingalgorithmwasusedinastudyofresistanthypertensioninthreehyper-tensionclinicsinAfrica,inastudyfundedbyGrandChallengesCanada.Thisapproachincreasedsystolicbloodpressurecontrolfrom25%inusualcareto75%inindividualizedcarebasedonaldosterone/reninprofiling.4
PrimaryAldosteronism Liddle’sVariants,AdducinPolymorphisms
RenalorRenovascular
Aldosterone High Low High
Renin Low Low High
Primarytreatment Aldosteroneantagonists:SpironolactoneEplerenone(Amilorideformenwhereeplerenoneisnotavailable)(rarelysurgery)
AmilorideAngiotensinreceptorblockersAliskiren(rarelyrevascularization)
Thepreciseandaccuratemeasurementofaldosteronebyenzymeimmunoassaycanbeanimportanttoolforthediagnosisoftheunderlyingcauseofhypertension,leadingtoappropriatetherapy.
Thisapproachnotonlyimprovesbloodpressurecontrol,thusreducingtheriskofstroke,heartfailureandrenalfailure,butalsoreducesadverseeffectsofmedicationandmayreducethecostofmedicationbyidentifyingspecifictherapy.
When this metabolism is altered, three patterns of aldosterone and renin activity levels can be produced:
PERFORMANCE
TheRfBinternationalcontrolswereassayedwiththeDBCELISAkit(CAN-ALD-500).Theresultsforthekitarethemean±SDof4independentexperiments(pg/mL).
TheresultsmatchbothLC-MS/MSresultsfromMayoClinicandfallwithintherangeofresultsestablishedfromallmethods.
RfB lot HM 2/15 A HM 2/15 BLC-MS/MS(MayoClinic) 570 110
LC-MS/MS(RfB)
Target
634 122
Range (16P–84P)
419–764 111–172
AllMethods(RfB)
Target
529 110
Range (16P–84P)
468–658 90–144
CAN-ALD-500(DBC) 636±70 105±16
Evaluation of International Controls
Aldosteroneconcentrationrangeinthepopulationdependsontheethnicandsocialcompositionandnutritionalfactors.
Each laboratory must determine its own reference ranges!
Aldosterone (pg/mL)
Freq
uenc
y
120 Putatively healthy individuals—UprightMedian:52pg/mL
Average:70pg/mL
95%confidencerange:ND–210pg/mL
REFERENCE RANGE
significantproportionofhypertensiveindividualssufferfromresistanthyper-
tensionduetofactorssuchasnon-compliance,poornutritionalpractice(highsalt,alcohol,licorice)orsecondaryhypertension(20%ofallresistanthypertensioncases).Diagnosingthecauseofhypertensionisofparamountimportancetoselectthecorrecttherapy.
Heartfailure,stroke,renalconditionsanddementiaaresomeofthecommon
consequencesofuncontrolledhyper-tension;theoccurrenceoftheseconditionshowever,canbereducedwhentheunderlyingsourceofresistanthypertensionisidentifiedandafollowuptherapyisapplied.
Renin
Angiotensinogen
Aldosterone
Angiotensin I
Angiotensin II
ACE
THE RENIN-ANGIOTENSIN-ALDOSTERONE AXISplaysakeyroleinresistanthypertension.
Duringnormalhomeostasis,reninisreleasedunderconditionsofdehydrationorlowbloodpressure(seefigurebelow).Renin enzymatic activitythenpromotesthecleavageofAngiotensinogenandgenerationofAngiotensinI,whichinturnistransformedintoAngiotensinIIandactivatesaldosteronerelease(whichcausessaltandwaterretention,andexcretionofpotassium,magnesium,andotherions).
A 1. SpenceJD.LessonsfromAfrica:theimportanceofmeasuringplasmareninandaldosteroneinresistanthypertension.Can J Cardiol.2012;28(3):254–7.
2. SpenceJD.Physiologictailoringoftherapyforresistanthypertension:20years’experiencewithstimulatedreninprofiling.Am J Hypertens.1999;12:1077–83.
3. SpenceJD.Physiologictailoringoftreatmentinresistanthypertension.Current Cardiology Reviews.2010;6:119–123.
4. JonesES,etal.PhysiologicallyIndividualizedTherapyforResistantHypertensioninAfrica.HypertensionTeachingSeminarOrganizedbytheInternationalSocietyofHypertension(ISH)AfricaRegionalAdvisoryGroupincollaborationwiththeEuropeanSocietyofHypertension(ESH),theInternationalForumforHypertensionControlandPreventioninAfrica(IFHA)andtheMozambicanHeartAssociation(AMOCOR);April18–19,2016;Maputo,Mozambique2016.
LITERATURE
new
at a glanceCatalogue number: CAN-ALD-500Number of test wells: 96Sensitivity: 9.1 pg/mLSample Volume: 50 µLTotal assay time: 80 mins.Validated against: LC-MS/MS
384 Neptune CrescentLondon, Ontario, Canada N6M 1A1
Tel: 519-681-8731 Fax: [email protected]
Manufacturing Innovative IVD for the World Since 1973
DBC’s NEW ALDOSTERONEELISA KIT INCLUDES AREADY-TO-USE CONJUGATE
AND BLOCKING AGENTS
REF CAN-ALD-500