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