1. 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. 2 Objectives For standardizations and specifications of the NAT and serological test For evaluation of reagents To achieve the safety of blood products

3. 3 Contents Virus :HBV, HCV, HIV-1 Fresh frozen plasma from voluntary donated bloods Source : Panels :100 panels each including negatives undiluted fresh frozen plasma from each donors Test Items :NAT and serological tests

4. 4 Test Items HBV DNA TaqMan PCR(J), AMPLICOR HBV Monitor(R) GenotypeDNA sequencing Subtypededuced from sequence IgM anti-HBcLumipulse(F) anti-HBsAUSAB(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. 5 ABCDFABCDF 11 25 55 1 HBV genotypeNumber early stage:51 late/carrier stage:42 HBV Panels HBV DNAHBsAganti-HBcNumberCategory Positive Negative Positive Negative Positive Negative Positive Negative 19 5 32 37 7 I II III IV V

6. 6 HBV Panels

7. 7

8. 8 Test Items HCV RNA TaqMan PCR(J), AMPLICOR HCV Monitor(R) GenotypePCR 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 AgLumipulse(F), HCV Ag ELISA(O), HCV Ag IRMA(O) Lumipulse(F)anti-HCV core RIBA(C)WB

9. 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. 10 Result False 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 results 9 with COBAS, 2 with MWP Improvement ==>> No false negative nor positive result in 11 sites in the follow-up survey

11. 11 Hisao Yugi Shigeru Tamatsukuri Norio Yahagi Tohru Yamada Hidetaka Minakawa Retsuji Yamanaka Shunji Iida Harumichi Matsukura Masaaki Mizui Mitsunobu Imai Teruhide Yamaguchi Hiroshi Yoshizawa Chief JRC Central B.C. Roche Diagnostics K.K. Ortho-Clinical Diagnostics K.K. Abbott Japan Co. Ltd. FUJIREBIO Inc. JRC Headquarter JRC 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