Implementation of an in-house quantitative ...
Title :
Implementation of an in-house quantitative real-time polymerase chain reaction method for Hepatitis B virus quantification in West African countries
Author(s) :
Ghosh, S. [Auteur]
Indian Institute of Technology Madras [IIT Madras]
Sow, A. [Auteur]
Institut de chimie des milieux et matériaux de Poitiers [UMR 7285] [IC2MP [Poitiers]]
Guillot, C. [Auteur]
Centre Hospitalier Universitaire [Strasbourg] [CHU Strasbourg]
Jeng, A. [Auteur]
Ndow, G. [Auteur]
Njie, R. [Auteur]
Toure, S. [Auteur]
Diop, Mame Andallah [Auteur]
Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 [LOG]
Mboup, S. [Auteur]
Kane, C.T. [Auteur]
Lemoine, M. [Auteur]
Institut d'Astrophysique de Paris [IAP]
Thursz, M. [Auteur]
Zoulim, F. [Auteur]
Mendy, M. [Auteur]
Chemin, I. [Auteur]
Indian Institute of Technology Madras [IIT Madras]
Sow, A. [Auteur]
Institut de chimie des milieux et matériaux de Poitiers [UMR 7285] [IC2MP [Poitiers]]
Guillot, C. [Auteur]
Centre Hospitalier Universitaire [Strasbourg] [CHU Strasbourg]
Jeng, A. [Auteur]
Ndow, G. [Auteur]
Njie, R. [Auteur]
Toure, S. [Auteur]
Diop, Mame Andallah [Auteur]
Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 [LOG]
Mboup, S. [Auteur]
Kane, C.T. [Auteur]
Lemoine, M. [Auteur]
Institut d'Astrophysique de Paris [IAP]
Thursz, M. [Auteur]
Zoulim, F. [Auteur]
Mendy, M. [Auteur]
Chemin, I. [Auteur]
Journal title :
Journal of Viral Hepatitis
Pages :
897-904
Publisher :
Wiley-Blackwell
Publication date :
2016-11
ISSN :
1352-0504
English keyword(s) :
France
Hepatitis
Hepatitis B
Hepatitis B virus
Infection
Patients
Time
Laboratories
London
Liver
Tenofovir
Viral Load
Polymerase Chain Reaction
Hepatitis
Hepatitis B
Hepatitis B virus
Infection
Patients
Time
Laboratories
London
Liver
Tenofovir
Viral Load
Polymerase Chain Reaction
HAL domain(s) :
Sciences du Vivant [q-bio]/Cancer
English abstract : [en]
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 ...
Show more >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 kitsShow less >
Show more >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 kitsShow less >
Language :
Anglais
Peer reviewed article :
Oui
Audience :
Internationale
Popular science :
Non
Source :
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