ArticleCureus2026
Performance Evaluation of the GeneNAT HHH Real-Time Polymerase Chain Reaction System for Simultaneous Detection of Hepatitis B Virus, Hepatitis C Virus, and Human Immunodeficiency Virus.
Article in Cureus, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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4 authors.
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Abstract
Background Hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) are major blood-borne pathogens responsible for significant global morbidity and mortality. Accurate and early diagnosis is essential for initiating timely treatment and reducing disease transmission. The World Health Organization has highlighted the need for diagnostic tools that are affordable, sensitive, specific, user-friendly, rapid, and robust, particularly in resource-limited settings. This study evaluates the clinical performance of a combined polymerase chain reaction (PCR)-based detection and viral load quantification assay for HBV DNA, HCV RNA, and HIV RNA (HHH assay) using the GeneNAT 300/340 Real-Time PCR System (Genetix Biotech Asia Pvt. Ltd., India) as the test assay. Its performance was compared with established reference assays: the COBAS® AmpliPrep/COBAS® TaqMan® v2.0 (Roche Diagnostics, USA) for HBV and HCV quantification as comparator assay 1 and the Xpert® HIV Viral Load assay (Cepheid, USA) for HIV RNA quantification as comparator assay 2. Methodology A retrospective analysis was conducted using 110 anonymized, de-identified archived plasma samples collected at a tertiary liver care center in India. The cohort comprised a study group of 53 confirmed viral load-positive samples (20 HBV, 18 HCV, and 15 HIV) and a control group of 57 samples negative for HBV, HCV, and HIV infection. For evaluation of diagnostic performance, all archived specimens were tested in parallel using the GeneNAT 300/340 Real-Time PCR System as test assay and the established reference assays: COBAS® AmpliPrep/COBAS® TaqMan® v2.0 for HBV and HCV viral load quantification as comparator assay 1, and the Xpert® HIV Viral Load assay for HIV RNA quantification as comparator assay 2. Results The GeneNAT 300/340 Real-Time PCR assay demonstrated 100% concordance with the comparator assays 1 and 2, with no false-positive or false-negative results observed. The assay achieved 100% sensitivity and specificity for all three infections and delivered results within approximately 60 minutes. Conclusions The GeneNAT 300/340 Real-Time PCR assay shows significant potential to strengthen diagnostic capacity in low- and middle-income countries. Its ease of use, rapid turnaround time, and robust performance make it a strong candidate for broader implementation, including screening in blood banks and peripheral healthcare settings as part of a decentralized diagnostic approach. However, larger-scale prospective studies are required to further establish its clinical utility, cost-effectiveness, and feasibility for widespread adoption.
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