Update from ISS on following EQA studies - NIBSC - … Cristiano...Biologicals Unit, CRIVIB Karen...

Karen Cristiano Biologicals Unit, CRIVIB

Update from ISS on following EQA studies: ISS EQA Tri-NAT (HCV, HIV and HBV) 2012

ISS EQA NAT HBV 2012

ISS EQA NAT WNV 2012

SOGAT XXIV

Ljubljana, 8-9 May 2013


2012 ISS EQAs for NAT

ISS EQA Tri-NAT (HCV, HBV, HIV)

ISS EQA HBV DNA

ISS EQA WNV RNA


Mandatory introduction of NAT

in Italian blood centers

June 2002 HCV RNA

April 2008 HIV RNA and HBV DNA


Italian NAT EQAs for

HCV, HIV and HBV

Since 2008, the National Blood Centre and the National

Centre for Immunobiologicals Research and

Evaluation (CRIVIB) of the ISS have been promoting

and organizing on an annual basis NAT EQAs for

HCV, HIV and HBV.


Italian Tri NAT EQAs - Scope

These programmes are aimed at assessing the performance of

laboratories in carrying out qualitative NAT assays for HCV

RNA, HIV RNA and HBV DNA…

… but they also provide participants with a tool to:

evaluate the effectiveness of their quality system by detecting

any possible weakness in the pre- and post-analytical steps of

the process

estimate the ability of their analytical methods to generate

results that meet the parameters and the technical

specifications of the assay


ISS EQA Tri NAT 2012 - Participants •

81 Italian blood centres

(+ 1 QC laboratory of a blood products’ manufacturer)

Estonia 1

Germany 2

Lithuania 1

Ireland 1

Slovenia 1

Spain 20

Switzerland 1

Thailand 16

43 foreign blood centres

Total no. of participants: 125


ISS EQA Tri NAT 2012 - NAT methods

Test kit Manufacturer Method 95% LOD IU/ml

HCV HIV HBV

COBAS TaqScreen Mpx

Roche Real Time PCR 10.7

49

3.7

COBAS TaqScreen Mpx v2.0

Roche

Real Time PCR 6.8

46

2.3

TMA/Ultrio Tigris

Novartis TMA 3

20.2

10.4

TMA/Ultrio Tigris Plus

Novartis TMA 3.1

27.6

2.1

TMA/Ultrio Manual

Novartis TMA 2.7

19.6

7.4

Virus Screening PCRa

GFE Blut Real Time PCR

1,770

1,011

179


ISS EQA Tri NAT 2012 -

Positive Materials

4th HCV RNA IS WHO 06/102 (genotype 1)

3rd HIV-1 RNA IS WHO 10/152 (subtype B)

2nd HBV DNA IS WHO 97/750 (genotype A)



Negative matrix

Starting material: human pooled plasma tested

negative for HCV, HIV and HBV (molecular and

serological markers)


ISS EQA Tri NAT 2012 - Panel composition

H I V

H C V

H B V

Phase 1 Phase 2 Phase 3 Phase 4

H B V

H B V

H I V

H B V

H I V

H C V

H C V

H C V

H I V

H I V

H B V

HCV RNA: two 60 IU/ml-samples and three 30 IU/ml-samples*

HIV-1 RNA: five 100 IU/ml-samples

HBV DNA: five 50 IU/ml-samples

Negative plasma: five samples

*These three samples were not used for participants’ evaluation

H C V


ISS EQA Tri NAT 2012 –

HCV results

IU/ml No. of samples

tested

Overall no. of

incorrect results

(only analytical

errors)

% Correct

results

60 250 1 (1) 99.6

30 372 22 (22) 94



HIV results


tested

Overall no. of

incorrect results

(only analytical

errors)

% Correct

results

100 621 2 (1) 99.7



HBV results


tested

Overall no. of

incorrect results

(only analytical

errors)

% Correct

results

50 622 1* (1) 99.8

*Sample tested positive for both HBV and HCV (COBAS TaqScreen Mpx v2.0)



Negative results

613 negative*/619samples tested

% correct results= 99.0

*6 failures: false positive results


The design of this EQA allowed participants to

verify their routine activity making them confident

regarding their performance, especially when the

level of molecular target is low (near or below the

detection limit of the NAT assay used)


Remarks


EQA: a five-year experience No. of non-proficient laboratories

2008 2009 2010 2011 2012

False negative results 6 2 4 6 2

False positive results 5 1 7 1 7

Pre/post-analytical errors 9 1 1 5 1

Total (%) 20/118 4/121 12/123 12/121 10/124 (16,95) (3,30) (9,76) (9,92) (8,06)




ISS EQA HBV DNA

ISS EQA WNV RNA


Test kit Manufacturer Method 95% LOD

IU/ml

No. of

labs

Cobas TaqMan HBV Test Roche Real Time PCR 12.0 1

Cobas TaqScreenMPX Test Roche Real Time PCR 3.7 15

Cobas TaqScreenMPX Test v2.0 Roche Real Time PCR 2.3 6

Procleix Ultrio Tigris Novartis TMA 10.4 9

Procleix Ultrio Plus Tigris Novartis TMA 2.1 6

Procleix Ultrio eSAS Novartis TMA 7.4 2

Procleix Ultrio Plus eSAS Novartis TMA 1.9 1

ISS EQA NAT HBV 2012 –

NAT methods

TOT. 40



Panel Composition

HBV DNA IS 97/750

(genotype A)

WHO Reference Panel

Paul Erlich Institute

code 5086/08

Genotype B (vial no. 5 of WHO Ref. Panel) diluted to ~ 36 UI/mL (24 ÷ 54)

Genotype C (vial no. 7 of WHO Ref. Panel) diluted to ~ 36 UI/mL (27 ÷ 47)

Genotype D (vial no. 10 of WHO Ref. Panel) diluted to ~ 36 UI/mL (26÷ 48)

IU/ml 216 36 36 6* 1* ~36 ~36 ~36 neg neg

*These samples were not used for participants’ evaluation


Test kit No.

labs

216

IU/ml

36

IU/ml

36

IU/ml

6

IU/ml

1

IU/ml

~ 36

IU/ml

~ 36

IU/ml

~ 36

IU/ml Neg

Cobas TaqMan HBV Test 1 100%

1/1

100%

1/1

100%

1/1

100%

1/1

0%

0/1

100%

1/1

100%

1/1

100%

1/1

100%

2/2

Cobas Taqscreen MPX Test 15 100%

15/15

100%

15/15

100%

15/15

93%

14/15

53%

8/15

100%

15/15

100%

15/15

100%

15/15

100%

30/30

Cobas Taqscreen MPX Test

v 2.0 6

100%

6/6

100%

6/6

100%

6/6

100%

6/6

66%

4/6

100%

6/6

100%

6/6

100%

6/6

100%

12/12

Procleix Ultrio Tigris 9 100%

9/9

100%

9/9

100%

9/9

100%

9/9

11%

1/9

100%

9/9

100%

9/9

100%

9/9

100%

18/18

Procleix Ultrio eSAS 2 100%

2/2

100%

2/2

100%

2/2

0%

0/2

0%

0/2

100%

2/2

100%

2/2

100%

2/2

100%

4/4

Procleix Ultrio Plus

(Tigris and eSAS) 7

100%

7/7

100%

7/7

100%

7/7

100%

7/7

29%

2/7

100%

7/7

100%

7/7

100%

7/7

100%

14/14

TOTAL 40 100%

40/40

100%

40/40

100%

40/40

92%

37/40

37%

15/40

100%

40/40

100%

40/40

100%

40/40

100%

80/80

ISS EQA NAT HBV 2012 – Results


Roche 22/22 22/22 21/22 12/22

Novartis 18/18 18/18 16/18 3/18

100 % 100 % 95.4 % 54 %

100 % 100 % 88 % 16 %

HBV DNA 216 IU/ml 36 IU/ml 6 IU/ml 1 IU/ml

Probit Analysis - 95% LOD (overall):

Roche 5.6 IU/mL

Novartis 8.4 IU/mL


Overall Performance


HBV DNA 216 IU/ml 36 IU/ml 6 IU/ml 1 IU/ml

Probit Analysis – 95% LOD (Ultrio)

Ultrio 10.7 IU/ml

Ultrio plus 3.3 IU/ml

ISS EQA NAT HBV 2012 -

Performance Ultrio

Ultrio 11/11 11/11 9/11 1/11

Ultrio plus 7/7 7/7 7/7 2/7

100 % 100 % 81.8 % 9.1 %

100 % 100 % 100 % 28.5 %


All participants correctly detected the 216 IU/ml-

samples (genotype A)

All participants correctly detected the 36 IU/ml-

samples (genotypes A, B, C and D)

Detection of the 6 and 1 IU/ml-samples reflected the

stated 95% LOD of the NAT assay used by the

participant

ISS EQA NAT HBV 2012 -

Conclusions




ISS EQA HBV DNA

ISS EQA WNV RNA


2 WNVD

1 WNVD

•21 WNVD

•17 WNF

• 14 NAT-positive

blood donors

4 WNVD

WNV in

Italy:


Current preventive measures applied

in Italy in the transfusion setting

Nationwide 28-day deferral of blood donors who

have spent at least one night in an affected area*

WNV-NAT testing of blood donations collected in

Italian risk areas (namely areas identified as affected

areas* in the previous year)

*Affected area: where a human case of WNND and/or a

WNV-NAT positive donor is confirmed


ISS EQA NAT WNV 2012 -

Panel composition

*Identical panel sent in July and in October (but different coding)

Total number of participants: 11

7 Laboratories: TMA

4 Laboratories: PCR

cp/ml 100 100 100 100 100 100 50 50 neg neg

Lineage 1 1 1 1 2 2 2 2

Phases 1

and 2*

Overall results: 100%


What’s in the pipeline?

New EQA for Tri NAT started in April 2013 (Phase I)

Deadline for registration: end of May 2013

New EQA NAT WNV starting in July 2013 (Phase I)

Deadline for registration: June 2013

These EQAs are open also to foreign laboratories and are

free of charge with the exception of samples’ shipment.

Contact Giulio Pisani at [email protected] to get the

instructions for a preliminary registration.

mailto:[email protected]


Thank you for your

attention!

Update from ISS on following EQA studies - NIBSC - … Cristiano...Biologicals Unit, CRIVIB Karen...

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