Preparation of plasmids containing HBV-full genome of genotype A to H and trial of HBV inactivation method Yoshiaki Okada Kiyoko Umemori Kazunari.

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Presentation transcript:

Preparation of plasmids containing HBV-full genome of genotype A to H and trial of HBV inactivation method Yoshiaki Okada Kiyoko Umemori Kazunari Yamaguchi National Institute of Infectious Disease

Background and Objects genotype B and C of HBV are dominant in Japan.Recently detection rate of genotype A is increasing in Tokyo. The preparations of HBV genotype panel are required to keep blood safety. Genotypes except A, B and C are difficult to collect in Japan. We have prepared cloned HBV-full genome by PCR. 1.Evaluation of NAT sensitivity among HBV genotypes. 2.Evaluation of HBs-Ag Diagnostic kits among HBV genotypes.

Design of Primers for HBV Full Genome Amplification (J.Virol.vol.69.p.5437-5444)

Panels of Plasmids containing HBV-full genome genotype cloning recombinant HBs-Ag sequence A done + done B done + done C done + done D done + done E done + done* F done + done G done + done Ｈ　　　 done + done *several bases are deleted.

Amplification efficiency among HBV genotypes Genotype A.B,C,D,and G genotype E and F

Evaluation of HBs-Ag diagnostic kits with recombinant HBs-Ag which were produced by prepared plasmids J Virological methods 136,254-256,2006

Development of estimation method of HBV inactivation HBV does not replicate in cell culture. Recently Owada reported that the treated HBV with neuraminidase enhance the binding to the cells which express asialoglycoprotein receptor .(J.Viral hepat.vol.13.11-18.2006)

Neuraminidase treatment enhance HBV infectivity ? HBV (+) HBV (+)　　　　　　　　　　　　　 neuraminidase ( - ) neuraminidase (+) Planning to quantitative assay

Conclusion 1.We have prepared plasmids containing HBV-full genome of genotype A to H respectively. 2.Prepared plasmid panel is useful to evaluate NAT and HBs-Ag detection kits.

Establishment of cell culture method of PrPres from BSE appendix Establishment of cell culture method of PrPres from BSE BSE (-) (+) I will report next SoGAT meeting 1 2 3 4 5 6 7 1.Neg. 2.X 1 3.X 102 4.X 103 5.X 104 6.X 105 7.X 106 Infected cells secreted infectious PrPres into the culture supernatant.