NAT Proficiency Study Program in Japan
Saeko Mizusawa, Yoshiaki OkadaNational Institute of Infectious Diseases, Japan
SoGAT XXIIn Brussels, Belgium
28-29 May 2009
BackgroundNational Standards for NAT Calibrated against WHO ISHCV(1999 ), HBV(2002 ), HIV(2002)
National Standards for NAT Calibrated against WHO ISHCV(1999 ), HBV(2002 ), HIV(2002)
Guideline for Industry on NAT 2004 HCV, HBV, HIV Validated method with 100 IU/ml (95%DL)
Guideline for Industry on NAT 2004 HCV, HBV, HIV Validated method with 100 IU/ml (95%DL)
Guideline for Look Back Study 2005Guideline for Look Back Study 2005 HCV: core antigenHCV: core antigen HBV: NATHBV: NAT HIV: antibodyHIV: antibody
Guideline for Look Back Study 2005Guideline for Look Back Study 2005 HCV: core antigenHCV: core antigen HBV: NATHBV: NAT HIV: antibodyHIV: antibody
Objectives
To assess the proficiency and sensitivity of the validated routine NAT implemented in plasma pool and blood screening.
To assess the proficiency and sensitivity of the validated routine NAT implemented in plasma pool and blood screening.
To assess the performance of diagnostic HBV-NAT to test recipients according to Look-back guideline.To chose the suitable HBV-NAT kits if necessary.
To assess the performance of diagnostic HBV-NAT to test recipients according to Look-back guideline.To chose the suitable HBV-NAT kits if necessary.
Materials and Methods
Panels :Panels :Prepared by NIID by diluting the respective national Standard. Each panel consists of 8 consists of 8 coded samples, 7 positive and one negative coded samples, 7 positive and one negative samples. samples.
Assay:Assay:Three Three independent assays on different days with validated routine NAT methodsNAT methods
Data :Data :Reported ”positive or negative” for qualitative assays and “concentration” for quantitative assays.The presented data were analyzed by NIID.
Panels :Panels :Prepared by NIID by diluting the respective national Standard. Each panel consists of 8 consists of 8 coded samples, 7 positive and one negative coded samples, 7 positive and one negative samples. samples.
Assay:Assay:Three Three independent assays on different days with validated routine NAT methodsNAT methods
Data :Data :Reported ”positive or negative” for qualitative assays and “concentration” for quantitative assays.The presented data were analyzed by NIID.
The 1st NAT Control Surveillance The 1st NAT Control Surveillance 20062006
HBV-NAT: ParticipantsHBV-NAT: Participants
Organization Location No. of Organizatio
ns
No. of Laboratorie
s
Blood Product Manufacturer& blood center
Japan 4 6US/EU 3 4
Diagnostic Laboratory
Japan 7 7
Kit Manufacturer
Japan 2 3
OCL Japan 1 1Total 17 21
AssayNumber of Assay Sets
Manufact.& blood c.
OCLDiagnostic
LabKit maker Total
Qualitative AmpliScreen 6 1 1 8 Monitor 1 1* 2 In-house (5 kinds )
7 2 9
TMA 1 1Total 15 1 4 20Quantitative Diagnostic kit Kit A 7 1* 8 Kit B 3 1 4 Total 10 2 12
Overall 15 1 10 6 31
NAT Control Surveillance 2006NAT Control Surveillance 2006HBV-NAT: MethodsHBV-NAT: Methods
HBV-NAT Control Surveillance 2006(1) Assays Required by NAT-Guideline
02468
101214161820
Nu
mb
er
of
As
sa
y S
ets
10000 3000 1000 300 100 30 10 0
HBV-DNA (IU/mL)
no positive/ 3 assays
1 positive/ 3assays
2 positive/ 3 assays
3 positive/ 3 assays
HBV-NAT Control Surveillance 2006(2) Assays Recommended by Guideline for Look-back Study
0
20
40
60
80
100
10000 3000 1000 300 100 30 10 0
HBV-DNA (IU/ mL)
Tota
l pos
itive
/ To
tal t
este
d
(%)
n=24 Diagnostic Kit A:Quantitative assay Used by all the 7 diagnostic labs.No false-positive results were observed.
Samples containing 100 IU/ml of HBV-DNA or higher were able to measur in 100%.
Diagnostic Kit A:Quantitative assay Used by all the 7 diagnostic labs.No false-positive results were observed.
Samples containing 100 IU/ml of HBV-DNA or higher were able to measur in 100%. Diagnostic Kit B:Quantitative assayUsed by major 3 diagnostic labs.No false-positive results were observed.
Samples containing 10000 IU/ml were able to measur in 100%, 3000 IU/ml were able to measur in 92%, 300 IU/ml or less were not measured.
Diagnostic Kit B:Quantitative assayUsed by major 3 diagnostic labs.No false-positive results were observed.
Samples containing 10000 IU/ml were able to measur in 100%, 3000 IU/ml were able to measur in 92%, 300 IU/ml or less were not measured.
The 2The 2ndnd NAT Control Surveillance NAT Control Surveillance 20072007
HCV-NAT & HIV-NAT: ParticipantsHCV-NAT & HIV-NAT: Participants
OrganizationLocation
HCV-NAT HIV-NATOrg Lab Org Lab
Blood Product Manufacturer& blood center
Japan 4 6 4 6US/EU 2 3 2 3
Diagnostic Lab
Japan 7 7 3 3
Kit Manufacturer Japan 1 1 1 1OCL Japan 1 1 1 1
Total 15 18 11 14
HCV-NAT Control Surveillance HCV-NAT Control Surveillance 20072007Methods (Qualitative)Methods (Qualitative)
Assay
Number of Assay Sets
Manufact.
& Blood C.
OCLDiagnostic
Lab
Kit make
rTotal
AmpliScreen 4 1 1 6
TaqScreen MPX
1 1
TMA Assay 1 1
In-house (4 kinds)
6 1 7
Total 12 1 2 15
Amplicap/Amplicor
4 1 5
COBAS Amplicor 2 2
TMA Assay 1 1
Total 7 1 8
Overall 12 1 7 3 23
HCV-NAT Control HCV-NAT Control Surveillance 2007Surveillance 2007
0
2
4
6
8
10
12
14
16
Nu
mb
er
of
As
sa
y S
ets
10000 1000 300 100 30 10 3 0
HCV- RNA (IU/mL)
no positive/ 3 assays
1 positive/ 3assays
2 positive/ 3 assays
3 positive/ 3 assays
44/4545/45
AssayNumber of Assay Sets
Manufacture
& Blood c.OCL
Diagnostic Lab
Kit maker Total
Qualitative AmpliScreen 4 1 1 6 TaqScreen 1 1 TMA 1 1 In-house (4 kinds )
6 1 7
小計 12 1 2 15Quantitative Monitor 2 1 3 Monitor Standard
1 1 2
小計 3 2 5 合計 12 1 3 4 20
HIV-NAT Assay Methods
HIV-NAT Control Surveillance
2007
0
2
4
6
8
10
12
14
16
Nu
mb
er
of
As
sa
y S
ets
10000 3000 1000 300 100 30 10 0
HIV-RNA (IU/mL)
no positive/ 3 assays
1 positive/ 3assays
2 positive/ 3 assays
3 positive/ 3 assays
44/45 37/45
Summary (1)NAT for Plasma Pool and Blood Screening
1. All the samples containing 1000 IU/ml virus or more were detectable: HBV in 60/60, HCV in 45/45, HIV in 45/45.No false positive results were observed.
2. Samples containing 300 IU/ml, which is three times the 95%DL , were positive for HBV in 60/60 (100%), for HCV in 45/45 (100%), for HIV in 44/45 (98%). The laboratory which failed to detect 1/3 of 300 IU/ml of HIV samples was able to detect 100 IU/ml in 2/3.
3. Samples containing 100 IU/ml (95%DL) were positive for HBV in 60/60 (100%), for HCV in 44/45 (98%), for HIV in 37/45 (82%).
4. Overall, the qualities of NAT, proficiency and the sensitivity , were satisfactory .
Summary ( 2 )NAT for Look-back Study
1. Two quantitative HBV-DNA kits were available (Kit A and B).No false positive results were observed in both kit A and kit B.
2. Kit A was able to measure all the positive samples (10 – 10000 IU/ml) while kit B was able to measure samples containing 3000 IU/ml or more.
Summary ( 3 )
NAT control surveillance should be continued:
To reflect the results on regulations.
To improve the performance of participants
To confirm that routine NAT is able to detect all the genotypes/subtypes.
To assess performance of newly developed assay systems.
Collaboration with:
Working Group on NAT Control SurveillanceChaired by Dr. Teruhide Yamaguchi National Institute of Health Sciences
Department of Bacteriology II, NIIDYoshinobu Horiuchi
Department of Blood and Safety Research, NIIDToshiaki Mizuochi
Kazunari Yamaguchi