1 Preparation of Standard Paneled Plasma for HBV, HCV, and HIV-1 in Japan Hisao Yugi Japanese Red Cross Central Blood Center Study Group of the Ministry.

1

Preparation of Standard Paneled Plasma for HBV, HCV, and HIV-1

in Japan

Hisao Yugi

Japanese Red Cross Central Blood Center

Study Group of the Ministry of Health, Labour and Welfare, Japan

2

Objectives

For standardizations and specifications of the NAT and serological test

For evaluation of reagents

To achieve the safety of blood products

3

Contents

Virus : HBV, HCV, HIV-1

Fresh frozen plasma fromvoluntary donated bloods

Source :

Panels : 100 panels each including negatives undiluted fresh frozen plasmafrom each donors

Test Items : NAT and serological tests

4

Test Items

HBV DNA TaqMan PCR(J), AMPLICOR HBV Monitor(R)

Genotype DNA sequencing

Subtype deduced from sequence

IgM anti-HBc Lumipulse(F)

anti-HBs AUSAB(A), Lumipulse(F)

anti-HBc AxSYM(A), Lumipulse(F), HBcAb ELISA(O)

DNA sequencingPre-C mutation

HBsAg PRISM(A), AxSYM(A), ARCHITECT(A),Lumipulse(F), Vitros(O), HBsAg ELISA(O)

Methods/Reagents

(A):Abbott, (F):FUJIREBIO, (J):JRC, (O):Ortho, (R):Roche

HBV Panels

5

ABCDF

11255511

HBV genotype Number

early stage: 51late/carrier stage: 42

HBV PanelsHBV DNA HBsAg anti-HBcNumberCategoryPositivePositivePositivePositiveNegative

NegativeNegativePositivePositiveNegative

NegativePositiveNegativePositiveNegative

195

32377

IIIIIIIVV

6

HBV Panels

7

HBV Panels

8

Test Items

HCV RNA TaqMan PCR(J), AMPLICOR HCV Monitor(R)

Genotype PCR

anti-HCV PHA(A), PA(F), Lumipulse(F), AxSYM(A), HCV Ab ELISA(O)

Methods/Reagents

(A):Abbott, (C):Chiron, (F):FUJIREBIO, (J):JRC, (O):Ortho, (R):Roche

HCV Panel

HCV core Ag Lumipulse(F), HCV Ag ELISA(O), HCV Ag IRMA(O)

Lumipulse(F)anti-HCV core

RIBA(C)WB

9

Preliminary Application Study

A nation-wide survey for HCV NAT

Coded panel for the survey- Genotype 1b, 2a, 2b (200-300 IU/mL)

- Negative plasma

Participants: - COBAS AMPLICOR : 92 HPs and Labs

- AMPLICOR (MWP) : 32 HPs and Labs

10

ResultFalse negative result

- 1b : 2.4% (3/124)

- 2a : 6.5% (8/124)

- 2b : 2.4% (3/124)

- Neg : 0.0% (0/124)

11 sites gave false negative results9 with COBAS, 2 with MWP

Improvement ==>> No false negative nor positive result in 11 sites in the follow-up survey

11

Hisao YugiShigeru TamatsukuriNorio YahagiTohru YamadaHidetaka MinakawaRetsuji YamanakaShunji IidaHarumichi MatsukuraMasaaki MizuiMitsunobu ImaiTeruhide YamaguchiHiroshi YoshizawaChief

JRC Central B.C.Roche Diagnostics K.K.Ortho-Clinical Diagnostics K.K.Abbott Japan Co. Ltd.FUJIREBIO Inc.JRC HeadquarterJRC HeadquarterJRC Osaka B.C.JRC Hiroshima B.C.Kanagawa Pref. Public Health Lab.National Inst. Health Sci.Hiroshima Univ. Graduate School Biomed. Sci.

Study group on the establishment of standardizations, specifications and test methods for the safety and quality of blood products under the Ministry of Health, Labour and Welfare, Japan