1. 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

2. 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.

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

4. 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.

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

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

7. 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 )

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

9. 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.

10. 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.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.