Performance of two real-time PCR assays for hepatitis B virus DNA detection and quantitation
Journal of Virological Methods
Laboratoire de Virologie, Centre Muraz, Bobo-Dioulasso, Burkina Faso; Unité VIH et Maladies Associées, Centre Muraz, Bobo-Dioulasso, Burkina Faso; INSERM U 1058, Infection by HIV and by agents with mucocutaneous tropism: from pathogenesis to prevention, 34394 Montpellier, France; Université Montpellier 1, 34090 Montpellier, France; CHU Montpellier, Département de Bactériologie-Virologie et Département d'Information Medicale, 34295 Montpellier, France; Africa Centre for Health and Population Studies, University of KwaZulu-Natal, Durban, South Africa
In-house developed real-time PCR (qPCR) techniques could be useful conjunctives to the management of hepatitis B virus (HBV) infection in resource-limited settings with high prevalence. Two qPCR assays (qPCR1 and qPCR2), based on primers/probes targeting conserved regions of the X and S genes of HBV respectively, were evaluated using clinical samples of varying HBV genotypes, and compared to the commercial Roche Cobas AmpliPrep/Cobas TaqMan HBV Test v2.0. The lower detection limit (LDL) was established at 104 IU/ml for qPCR1, and 91 IU/ml for qPCR2. Good agreement and correlation were obtained between the Roche assay and both qPCR assays (r = 0.834 for qPCR1; and r = 0.870 for qPCR2). Differences in HBV DNA load of > 0.5 Log10 IU/ml between the Roche and the qPCR assays were found in 49/122 samples of qPCR1, and 35/122 samples of qPCR2. qPCR1 tended to underestimate HBV DNA quantity in samples with a low viral load and overestimate HBV DNA concentration in samples with a high viral load when compared to the Roche test. Both molecular tools that were developed, used on an open real-time PCR system, were reliable for HBV DNA detection and quantitation. The qPCR2 performed better than the qPCR1 and had the additional advantage of various HBV genotype detection and quantitation. This low cost quantitative HBV DNA PCR assay may be an alternative solution when implementing national programmes to diagnose, monitor and treat HBV infection in low- to middle-income countries where testing for HBV DNA is not available in governmental health programmes. © 2014 Elsevier B.V.
virus DNA; hepatitis B surface antigen; hepatitis B virus X protein; oligonucleotide probe; primer DNA; transactivator protein; virus DNA; article; controlled study; DNA determination; DNA extraction; DNA virus; genotype; Hepatitis B virus; human; income; limit of detection; major clinical study; polymerase chain reaction; prevalence; priority journal; real time polymerase chain reaction; virus load; comparative study; evaluation study; genetics; hepatitis B; Hepatitis B virus; isolation and purification; molecular diagnosis; oligonucleotide probe; procedures; real time polymerase chain reaction; DNA Primers; DNA, Viral; Hepatitis B; Hepatitis B Surface Antigens; Hepatitis B virus; Humans; Molecular Diagnostic Techniques; Oligonucleotide Probes; Real-Time Polymerase Chain Reaction; Trans-Activators; Viral Load