Skip to Main content Skip to Navigation
Journal articles

Implementation of an in-house quantitative real-time polymerase chain reaction method for Hepatitis B virus quantification in West African countries

Abstract : Hepatitis B virus (HBV) is a major cause of chronic liver disease worldwide. HBV infection is diagnosed by serological tests, while real-time polymerase chain reaction (qRT-PCR) assays are used to quantify viral load, which is a crucial parameter to determine viral replication and to monitor antiviral treatments. However, measuring viral load in resource-limited countries remains nonsystematic, due to the high cost of commercial kits. Here, we describe the development, validation and implementation of a low-cost, in-house qRT-PCR assay to monitor HBV viral load in chronic carriers enrolled in the PROLIFICA programme in the Gambia and Senegal. Over 1500 HBsAg-positive patients, including 210 chronically infected HBV patients, who were given antiviral treatment (tenofovir), were monitored by qRT-PCR using the SYBR Green- and HBV-specific primers. Twenty-four tenofovir-treated patients were followed up and their viral load was tested every 3 months over the 12-month experimental time course. Compared to commercial assays, our in-house assay was shown to be (i) highly reliable, with good intra- and interassay reproducibility over a wide range (45-4.5 x 108 copies mL-1 ), (ii) very similar in the viral loads detected (R2 = .90), (iii) highly sensitive, as it detected loads as low as 30 copies mL-1 (~5 IU mL-1 ), (iv) cheaper (2- to 3-fold), (v) easier to implement and (vi) more rapid. Based on our experience, we recommend this assay as a reliable alternative to commercial assays, for monitoring HBV viraemia in resource-limited, highly endemic countries to reduce the cost and technical obstacles associated with commercial kits
Document type :
Journal articles
Complete list of metadata

https://hal.archives-ouvertes.fr/hal-01792611
Contributor : Christelle Cheval Connect in order to contact the contributor
Submitted on : Tuesday, May 15, 2018 - 3:59:19 PM
Last modification on : Friday, June 3, 2022 - 10:22:03 AM

Links full text

Identifiers

Citation

S. Ghosh, A. Sow, C. Guillot, A. Jeng, G. Ndow, et al.. Implementation of an in-house quantitative real-time polymerase chain reaction method for Hepatitis B virus quantification in West African countries. Journal of Viral Hepatitis, Wiley-Blackwell, 2016, 23, pp.897-904. ⟨10.1111/jvh.12561⟩. ⟨hal-01792611⟩

Share

Metrics

Record views

55