Assessment of semi-automated nucleic acid testing programme in a Regional Blood Transfusion Centre

Abstract

Background: Detection of human immunodeficiency virus type-1 (HIV-1), hepatitis-C (HCV) and hepatitis-B virus (HBV) in the blood donors is crucial. An efficient form of detection is nucleic acid testing (NAT) in blood screening. We assessed the suitability of commercial NAT testing in a developing country, focusing on the Altona RealStar assay and the method of Sacace Biotechnologies.

Methods: We have standardised and validated commercially available NAT kits with a semi-automated system for detection of HBV, HCV and HIV-1 in blood donations. The MP-NAT (mini-pool) assay consists of pooling of sample, virus extraction, amplification and detection with commercially available NAT kits. An internal control (IC) is incorporated in the assay to monitor the extraction, target amplification and detection process.

Results: The sensitivity of the Altona RealStar assay at 10-MP for each viral target was evaluated, HBV showed amplification in all diluted positive samples of 100, 50, 25, 10 and 5 IU/ml. HIV and HCV infected samples showed amplification in all diluted positive samples of 500, 100, 50 and 30 IU/ml. For HIV, out of six diluted samples of 30 IU/ml, five were amplified. A total of 14,170 seronegative blood samples were tested by RealStar PCR kit in 10-MP and 6 (0.042%) samples/pools were positive. A total of 65,362 seronegative blood donations were also tested by kits of Sacace Biotechnologies, in 10-MP and 45 (0.075%) pools were positive. The prevalence of combined NAT yield cases among routine donors was 1 in 1559 donations tested for all the 3 viruses.

Conclusion: The semi-automated combined system for NAT screening assays is robust, sensitive, reproducible, and this gives an additional layer of safety with affordable cost.

Keywords:

• Nucleic acid testing (NAT)
• HIV
• HBV
• HCV
• Semi-automated
• seronegative

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Acknowledgements

We would like to offer our sincere thanks to Dr Anand Deshpande, P. D. Hinduja National Hospital, Mumbai, India for providing the assistance with ID-NAT positive and negative plasma samples for quality-control programme. The authors thank all individuals from the TTI department of the SRK&RC, who were involved in doing the ELISA of the blood donors. We also acknowledge Dr N. Vasavada, SRK&RC, Deputy Director, who has worked tirelessly for making the NAT testing project viable.

Disclosure statement

No potential conflict of interest was reported by the authors.