External quality assessment for the detection of HCV RNA, HIV RNA and HBV DNA in plasma by nucleic acid amplification technology: a novel approach.
ABSTRACT In this EQA study a novel approach was used to assess the performance of blood centres and blood product manufacturers in detecting the possible contamination of plasma with HCV, HIV and HBV by NAT.
A panel of 12 samples, three negative and three positive for each virus, was distributed to the EQA participants. The positive samples were prepared, using the respective WHO standards, in order to obtain a viral concentration of about three times the 95% DL of the methods most commonly used by laboratories involved in blood screening by NAT. Participants were requested to test each sample of the panel on different days, possibly by different operators using their routine NAT assay.
Overall, the participants' performance was satisfactory. In particular, 49 of the 59 participants (83%) were able to correctly identify all samples. Regarding the remaining 10 laboratories, in three cases a deviation from the laboratory's procedure that could be attributed to an operator's mistake was observed, in two cases a possible cross-contamination occurred while in the remaining five cases the failure to detect the positive samples couldn't be ascribed to any relevant deviation in the laboratory's procedure.
The novel design of this EQA study allowed participants to verify their day by day activity as the study was carried out in the context of their routine testing. Under these conditions, it was demonstrated that, despite the high level of automation reached by NAT assays, human errors can still occur.
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ABSTRACT: Since 1 July 1999, in accordance with European regulations, only batches of blood products obtained from plasma pools tested and found to be non-reactive for hepatitis C virus (HCV) RNA are being released. As monitoring the performance of manufacturers involved in plasma pool testing is important to ensure reliable amplification techniques, the Istituto Superiore di Sanità, as the Italian regulatory authority, organized an external quality assessment study. A reference HCV RNA panel calibrated in international units (IU) was sent to each participant to be tested according to the validated procedure they routinely used in plasma pool testing. The panel consisted of 20 coded samples, four of which were obtained from a negative plasma pool. The remaining 16 samples, prepared by diluting the national reference preparation (ISS HCV RNA 0498), represented four half-log dilution series, each consisting of four samples containing 100, 32, 10 and 3.2 IU/ml of HCV RNA. The overall performance of the laboratories was very satisfactory. All laboratories correctly identified the negative samples. The 100- and 32-IU/ml samples were both detected in 98.4% of the assays, while the 10- and 3.2-IU/ml samples were detected in 73.4 and 50.0% of the assays, respectively. No substantial differences were observed between in-house procedures and commercial kits. This external quality assessment study showed that manufacturers of blood products have reached a high level of proficiency that fully complies with the European Pharmacopoeia requirements. This finding is reassuring in the context of the safety of blood products.Vox Sanguinis 11/2001; 81(3):143-7. · 2.85 Impact Factor
- Vox Sanguinis 06/2002; 82(4):211-2. · 2.85 Impact Factor
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ABSTRACT: A collaborative study was undertaken to establish a replacement for the current (1st) World Health Organization (WHO) hepatitis C virus (HCV) International Standard, 96/790. Both the 1(st) International Standard and the replacement standard were prepared from the same starting material by diluting a high titre genotype 1a HCV isolate in pooled, human plasma. The only difference was that each standard was lyophilized in two, separate lyophilisation runs but under the same conditions. In the study to establish the 1st International Standard, no significant difference in potency was found between the material eventually designated as the 1st International Standard and that now selected as the 2nd International Standard. The present study also showed no significant differences between the materials stored at -20 degrees C and no evidence of degradation over 5 years. Material 96/798 was established as the 2nd HCV International Standard and assigned the same unitage as the 1st International Standard, i.e. 10(5) IU/ml (50,000 IU/vial).Vox Sanguinis 05/2005; 88(3):202-4. · 2.85 Impact Factor