Yi-Chen Yang, Yu-Hsuan Chen, Show-Lan Chiu, Hwei-Fang Cheng

Drug Biology Division, Bureau of Food and Drug Analysis

Department of Health, Taiwan, ROC

National Standards for Blood Viruses in Taiwan

SoGAT XXI May 2009

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• To improve the safety of plasma products – NAT requirements in Taiwan (2002)

• The plasma pool or mini-pool should be HBV NAT (-), HCV NAT (-), and HIV NAT (-)

• For S/D treated blood products, one additional VI/R step should be performed, or the plasma pool should be HAV NAT (-)

• B19 NAT on plasma pool or mini-pool is suggested

~ the cut-off limit of B19 DNA should be < 105 IU/mL

Current Status of NAT Regulations

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Class Ilow risk

Class IIhigher risk

Class III highest risk

• Most exempt from the Medical Device GMP/QSD

• Sterile products shall still comply with Medical Device GMP/QSD

Medical Device GMP/QSD

Medical Device GMP/QSD

Registration and Market approval

Registration and Market approval

Registration and Market approval*

Regulatory Framework of Medical Device

* Special Requirements for Class III IVD ~ Submit sample to BFDA for licensing tests

The current focus of BFDA’s work in biological standards is for IVDs that are either used in blood screening or plasma pool testing for further fractionation process.

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• High titer HBV DNA (+) plasma– Screening for other blood viruses– Quantitative analysis and genotyping

• Diluted with pooled human plasma*– *Screening for all major blood viruses

• Calibrate the titers of candidates against the IS (97/746) by a collaborative study

– 11 Labs from 7 countries• Official Medicine Control Laboratories (OMCL) & CDC (Taiwan) • Hospital laboratory & Reference laboratory • Blood Service• Manufacturers of in vitro diagnostics

• Stability study 4 , -20 , -80 ℃ ℃ ℃

HBV DNA National Standard and Working Reagent

ProductItem HBV DNA National Standard HBV DNA Working Reagent

Conc. (IU/mL) 106 103

Genotype genotype B genotype B

Lot number BFDA lot 92-08 BFDA lot 92-08W

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Exclude the data out of the range of mean ± 2 SD

HBV DNA standard HBV DNA standard

HBV DNA working reagent

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HCV RNA National Standard and Working Reagent

ProductItem HCV RNA National Standard HCV RNA Working Reagent

Conc. (IU/mL) 5.2 × 104 890

Genotype genotype 1b genotype 1b

Lot number BFDA lot 93-09 BFDA lot 93-09W

• High titer HCV RNA (+) plasma– Screening for other blood viruses– Quantitative analysis and genotyping

• Diluted with pooled human cryosupernatant*– *Screening for all major blood viruses

• Calibrate the titers of candidates against the IS (96/790) by a collaborative study

– 8 Labs from 6 countries• Official Medicine Control Laboratories (OMCL) & CDC (Taiwan) • Hospital laboratory & Reference laboratory• Blood Service• Manufacturers of in vitro diagnostics

• Stability study 4 , -20 , -80 ℃ ℃ ℃

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Exclude the data out of the range of mean ± 2 SD

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B19 DNA National Standard and Working Reagent

ProductItem B19 DNA National Standard B19 DNA Working Reagent

Conc. (IU/mL) 1.9 × 106 2.0 × 104

Genotype genotype 1 genotype 1

Lot number BFDA lot 94-08 BFDA lot 94-08W

• High titer B19 DNA (+) plasma– Screening for other blood viruses– Quantitative analysis and genotyping

• Diluted with pooled human cryosupernatant*– *Screening for all major blood viruses

• Calibrate the titers of candidates against the IS (99/800) by a collaborative study

– 10 Labs from 7 countries• Official Medicine Control Laboratories (OMCL)• NAT testing laboratory • Manufacturers of plasma products• Manufacturers of in vitro diagnostics

• Stability study 25 , 4 , -20 , -80 ℃ ℃ ℃ ℃

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All data were within the range (mean ± 2 SD), showed that all laboratories are in good agreement with the results.

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National Standards and Reference Panels for Serological AssaysNational Standards and Reference Panels for Serological Assays

No. Item Lot Note

1 1st National Standard for HBsAg (subtype ad)* 89-06 106 IU/mL

2 Anti-HCV Reference Panel 92-01

3 HBsAg Sensitivity Panel 94-12

4 Anti-HIV-1 Reference Panel 96-11

National Standards and Working Reagents National Standards and Working Reagents for for NATNAT Assays Assays

No. Item Lot Note

1 1st National Standard for HBV DNA* 92-08 106 IU/mL

2 1st Working Reagent for HBV DNA* 92-08W 103 IU/mL

3 1stNational Standard for HCV RNA* 93-09 5.2×104 IU/mL

4 1st Working Reagent for HCV RNA* 93-09W 890 IU/mL

5 1st National Standard for Parvovirus B19 DNA 94-08 1.9×106 IU/mL

6 1st Working Reagent for Parvovirus B19 DNA 94-08W 2×104 IU/mL

• Candidate Standard for HIV-1 RNA being formulated

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Anti-HIV-1 Reference Panel

• Anti-HIV-1 (+) plasma selection– Screening for other blood viruses– Characterized with anti-HIV diagnostic kits, HIV viral-load kit

• Panel members characterization by a collaborative study– 12 Labs from 5 countries

• Official Medicine Control Laboratories (OMCL) & CDC (Taiwan) • Hospital laboratory & Reference laboratory• Blood Service• Manufacturers of in vitro diagnostics

– Include 22 anti-HIV diagnostic kits & Lab. in-house WB assay

• Stability study 25 , 4 , -20 , -80 ℃ ℃ ℃ ℃

• The panel contains 6 characterized specimens– Include strong-reactive, weak-reactive, and negative samples

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• Class III IVD licensing test (BFDA)

• Post-market surveillance for high risk class III IVD (BFDA) – HBsAg diagnostic kits (2008)

• Pick up 2 products fail to meet the sensitivity requirement– Anti-HCV diagnostic kits (2009)

• kits receiving test in blood service & hospital lab. (IVD manufactures)

• R&D and quality control for diagnostic kits (IVD/Biotech manufactures)

• Sensitivity evaluation for purchasing diagnostic kits (Taiwan CDC)

• Research (BFDA & Hospital lab.)

Current Status of National Standards Used in Taiwan

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Quantitative Data from Users (I)

HBV STD (Lot 92-08)

Lab-Sample code

Log IU/mL

106 IU/mL TCD-BAbm1 5.699

TCD-BAbm2 5.580

TCD-BAbm3 5.505

TCD-BRoT1 6.041

TCD-BRoT2 6.215

BFD-BRoT1 6.371

HBV WR (Lot 92-08W)

Lab-Sample code

Log IU/mL

103 IU/mL TCD-BWAbm1 3.146

TCD-BWRoT1 3.326

Med-BWRoT1 3.049

Med-BWRoT2 3.021

Med-BWRoT3 3.196

Med-BWRoT4 3.253

Med-BWRoT5 3.212

Med-BWRoT6 3.161

Med-BWRoT7 3.299

BFD-BWRoT1 3.146

BFD-BWRoT2 3.079

BFD-BWRoT3 3.255

BFD-BWRoT4 3.079

BFD-BWRoT5 3.176

• Collaborative study resultMean ± 2SD: 6 ± 0.38 Log IU/mL

• Collaborative study resultMean ± 2SD: 3 ± 0.44 Log IU/mL

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Quantitative Data from Users (II)

HCV STD (Lot 93-09)

Lab-Sample code

Log IU/mL

5.2 ×104 U/mL TCD-CAbm1 4.199

TCD-CAbm2 3.700

TCD-CAbm3 4.155

TCD-CRoT1 4.869

TCD-CRoT2 4.913

TCD-CRoT3 5.045

HCV WR (Lot 93-09W)

Lab-Sample code

Log IU/mL

890 IU/mL TCD-CWAbm1 2.389

TCD-CWRoT1 3.243

Reg-CWRoT1 2.807

Reg-CWRoT2 2.455

Med-CWRoT1 2.799

Med-CWRoT2 2.736

Med-CWRoT3 2.732

Com-CWRoT1 2.600

Com-CWRoT2 2.630

• Collaborative study resultMean ± 2SD: 4.719 ± 0.192 Log IU/mL

• Collaborative study resultMean ± 2SD: 2.949 ± 0.486 Log IU/mL

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• We have established HBV, HCV, and B19 national standards and working reagents for NAT assays.

– The standards were prepared by BFDA, and established through the international collaborative studies.

• Candidate standard for HIV-1 RNA being formulated.

• These NAT standards are currently used for evaluation of the molecular diagnostics market in Taiwan.

– Licensing test & post-market surveillance for high risk class III IVD

– Kits receiving test in blood service & hospital lab

– R&D and quality control for diagnostic kits

– Sensitivity evaluation for purchasing diagnostic kits

Summary

Thank you for your attention

AcknowledgementsThanks to all participants

of the collaborative study groups

Thanks to all data-feedback users