1. Establishment of National Standards for NAT Blood Viruses and NAT Proficiency Study Program in Japan
Saeko Mizusawa, Yoshiaki Okada
National Institute of Infectious Diseases, Japan
SoGAT XX
In Warsaw, Poland
12-13 June 2007

2. Study Design for National NAT Standards
<Preparation of the Candidate: JRC>
A candidate plasma was diluted in cryosupernatant, and stored at –80℃.
<Organizer & Participants>
NAT Working Group (Chaired by Dr Teruhide Yamaguchi )
Industries for Plasma Derivatives/ JRC/Proficient Labs/NIID
< Assay >
End-point assay
1st assay: fold dilutions to determine endpoint
2nd –5th assay: 7 half-log dilutions around the endpoint
<Potency of the Candidate >
Calibrated against WHO International Standards
Subcommittee on Safety for Plasma-Derived Products
Working Group on the Establishment of National Standard for Nucleic Acid Technology Assay
<Preparation of the Candidate>
2 Candidate Source Plasma (>10 6 IU/mL)
The selected plasma was diluted to 105IU per mL,-80C
<Assay for Determination of the Potency >
End-point assay
Five independent assay on different days
Five independent assays on different days
1st assay: fold dilution to determine endpoint
2nd –5th assay: 7 half-log dilution around the endpoint
Candidate (fresh vial )
WHO IS (reconstitute & distribute in aliquots, stored at –80C)
Diluted in plasma
<Estimate of the Potency for the Candidate>

3. Summary of Assays Used for HCV-NAT by the Participants in the Study
Table1. Assays used in the collaborative study.
a) R&D:Smi-test EX-R&D
Amplicor: Amplicor HCV virsion 1 (Roche)
QIAamp:QIAamp DNA Blood Mini Kit (QIAGEN)
b) Eq. Vol. Amplified : the equivalent volume of sample that was amplified in an assay

4. Potency of HCV-122 Calibrated against WHO IS for HCV-RNA (96/790)
Overall (a) = the overall mean potency calculated from the all laboratories. Overall (b) = the overall mean potency calculated from the data excluding laboratories 1 and 2.
Overall (a) = the overall mean potency calculated from the all laboratories
Overall (b) = the overall mean potency calculated from the data excluding laboratories 1 and 2.

5. Potency of HCV-122 Calibrated against WHO IS for HCV-RNA (96/790)
Mean ( )
Anti-log (0.979 – 1.588)

6. Potency of HBV-129 Calibrated against the WHO IS for HBV-DNA (97/746)3 4
5-1
5-2 6 7 2 -3 -2 -1
Lab. Code No.
Log Rerative Potency to
IS 97/746
Mean ( )
Anti-log ( )
Relative Potency against WHO IS
Potency
for HBV-DNA (97/746)
( IU/mL)
Log
-0.352 (
-0.165) 10
5.65
Anti-log
0.444 (
-0.683)
4.4x10 5

7. Potency of HIV-00047 Calibrated against the WHO IS for HIV-DNA (97/656)
1.8 temes 10 to the fifth

8. National Standards for NAT, Japan
International standard
HCV
(genotype)
1.0x105
( 1b )
( 1 )
HBV
4.4x105
( C )
1.0x106
( A )
HIV-1
1.4x105
( B )
IU/mL
Each vial of a National Standard contains
0.5ml of positive plasma diluted incryosupernatant and should be stored at –80℃.

9. NAT Proficiency Program in Japan
National Standard for NAT
1999 HCV
2002 HBV
2002 HIV
Guideline for industry on NAT 2004
HCV, HBV, HIV
detection limit: at least 100IU/mL
NAT Control Surveillance
2006 HBV
2007 HCV
2007 HIV

10. HBV-NAT Control Surveillance 2006
Panel: 3 independent assays
National Standard for HBV-NAT (Genotype C)
10000、3000、1000、300、100、30、10　IU/mL & HBV Negative Plasma
Laboratories of Industries :
Location
No. of
Industries
Laboratories
Japan 4 6
USA 2 EU 1
Total 7 10

11. Results of HBV-NAT Control Surveillance
36 assays =3 assays x 12 assay systems
% of Positive results

12. Summary National Standard for NAT
1999 HCV
2002 HBV
2002 HIV
HBV-NAT for Blood Products used in Japan Fulfills Sensitivity Required in Japan NAT Guideline

13. Collaboration with: Department of Bacteriology II, , NIID
Yoshinobu Horiuchi
Department of Blood and Safety Research, NIID
Mizuochi Toshiaki
Kazunari Yamaguchi
Working Group on Establishment of the National Standards for Nucleic Acid　Technology Assay
Working Group on NAT Control Surveillance