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
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 43 foreign blood centres Estonia 1 Germany 2 Lithuania 1 Ireland 1 Slovenia 1 Spain 20 Switzerland 1 Thailand 16 Total no. of participants: 125 (+ 1 QC laboratory of a blood products manufacturer)
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 PCR a GFE Blut Real Time PCR 1,770 1,011 179
ISS EQA Tri NAT 2012 - Positive Materials 4 th HCV RNA IS WHO 06/102 (genotype 1) 3 rd HIV-1 RNA IS WHO 10/152 (subtype B) 2 nd HBV DNA IS WHO 97/750 (genotype A)
ISS EQA Tri NAT 2012 - 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
ISS EQA Tri NAT 2012 HIV results IU/ml No. of samples tested Overall no. of incorrect results (only analytical errors) % Correct results 100 621 2 (1) 99.7
ISS EQA Tri NAT 2012 HBV results IU/ml No. of samples 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)
ISS EQA Tri NAT 2012 Negative results 613 negative*/619samples tested % correct results= 99.0 *6 failures: false positive results
ISS EQA Tri NAT 2012 - Remarks 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)
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)
2012 ISS EQAs for NAT ISS EQA Tri-NAT (HCV, HBV, HIV) ISS EQA HBV DNA ISS EQA WNV RNA
ISS EQA NAT HBV 2012 NAT methods Test kit Manufacturer Method 95% LOD IU/ml Cobas TaqMan HBV Test Roche Real Time PCR 12.0 1 No. of labs 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 TOT. 40
ISS EQA NAT HBV 2012 Panel Composition IU/ml 216 36 36 6* 1* ~36 ~36 ~36 neg neg HBV DNA IS 97/750 (genotype A) *These samples were not used for participants evaluation 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)
ISS EQA NAT HBV 2012 Results 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 1/1 1/1 1/1 1/1 0% 0/1 1/1 1/1 1/1 2/2 Cobas Taqscreen MPX Test 15 15/15 15/15 15/15 93% 14/15 53% 8/15 15/15 15/15 15/15 30/30 Cobas Taqscreen MPX Test v 2.0 6 6/6 6/6 6/6 6/6 66% 4/6 6/6 6/6 6/6 12/12 Procleix Ultrio Tigris 9 9/9 9/9 9/9 9/9 11% 1/9 9/9 9/9 9/9 18/18 Procleix Ultrio esas 2 2/2 2/2 2/2 0% 0/2 0% 0/2 2/2 2/2 2/2 4/4 Procleix Ultrio Plus (Tigris and esas) 7 7/7 7/7 7/7 7/7 29% 2/7 7/7 7/7 7/7 14/14 TOTAL 40 40/40 40/40 40/40 92% 37/40 37% 15/40 40/40 40/40 40/40 80/80
ISS EQA NAT HBV 2012 Overall Performance HBV DNA 216 IU/ml 36 IU/ml 6 IU/ml 1 IU/ml Roche 22/22 22/22 21/22 12/22 100 % 100 % 95.4 % 54 % Novartis 18/18 18/18 16/18 3/18 100 % 100 % 88 % 16 % Probit Analysis - 95% LOD (overall): Roche Novartis 5.6 IU/mL 8.4 IU/mL
ISS EQA NAT HBV 2012 - Performance Ultrio HBV DNA 216 IU/ml 36 IU/ml 6 IU/ml 1 IU/ml Ultrio 11/11 11/11 9/11 1/11 100 % 100 % 81.8 % 9.1 % Ultrio plus 7/7 7/7 7/7 2/7 100 % 100 % 100 % 28.5 % Probit Analysis 95% LOD (Ultrio) Ultrio Ultrio plus 10.7 IU/ml 3.3 IU/ml
ISS EQA NAT HBV 2012 - Conclusions All participants correctly detected the 216 IU/mlsamples (genotype A) All participants correctly detected the 36 IU/mlsamples (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
2012 ISS EQAs for NAT ISS EQA Tri-NAT (HCV, HBV, HIV) ISS EQA HBV DNA ISS EQA WNV RNA
WNV in Italy: 4 WNVD 21 WNVD 17 WNF 14 NAT-positive blood donors 1 WNVD 2 WNVD
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 Phases 1 and 2* cp/ml 100 100 100 100 100 100 50 50 neg neg Lineage 1 1 1 1 2 2 2 2 *Identical panel sent in July and in October (but different coding) Total number of participants: 11 7 Laboratories: TMA 4 Laboratories: PCR Overall results:
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 veq.nat@iss.it to get the instructions for a preliminary registration.
Thank you for your attention!