Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia

Maimuna E Mendy; Steve Kaye; Marianne van der Sande; Pura Rayco-Solon; Pauline A Waight; Deborah Shipton; Dorka Awi; Paul Snell; Hilton C Whittle; Samuel J McConkey

doi:10.1186/1743-422X-3-23

Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia

Maimuna E Mendy, Steve Kaye, Marianne van der Sande, Pura Rayco-Solon, Pauline A Waight, Deborah Shipton, Dorka Awi, Paul Snell, Hilton C Whittle, Samuel J McConkey

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

BACKGROUND/AIM: The study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials.

METHOD: Quantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers.

RESULTS: The qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 x 103 to 1.5 x 108 copies/mL) and correlated well with those from a commercial assay (r = 0.91, (p < 0.001). Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% (161/209) of HBeAg negative and in all HBeAg positive carriers.

CONCLUSION: This method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.

Original language	English
Pages (from-to)	23
Journal	Virology Journal
Volume	3
DOIs	https://doi.org/10.1186/1743-422X-3-23
Publication status	Published - 4 Apr 2006

Keywords

Adolescent
Adult
Age Factors
Aged
Antiviral Agents
Carrier State
Child
Child, Preschool
DNA Primers
DNA, Viral
Gambia
Hepatitis B Antibodies
Hepatitis B Surface Antigens
Hepatitis B e Antigens
Hepatitis B virus
Hepatitis B, Chronic
Humans
Infant
Lamivudine
Male
Middle Aged
Polymerase Chain Reaction
Reproducibility of Results
Reverse Transcriptase Inhibitors
Sensitivity and Specificity
Viral Load
Journal Article

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10.1186/1743-422X-3-23

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@article{e2add5caeb0a426296e01ff2e8d8c847,

title = "Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia",

abstract = "BACKGROUND/AIM: The study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials.METHOD: Quantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers.RESULTS: The qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 x 103 to 1.5 x 108 copies/mL) and correlated well with those from a commercial assay (r = 0.91, (p < 0.001). Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% (161/209) of HBeAg negative and in all HBeAg positive carriers.CONCLUSION: This method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.",

keywords = "Adolescent, Adult, Age Factors, Aged, Antiviral Agents, Carrier State, Child, Child, Preschool, DNA Primers, DNA, Viral, Gambia, Hepatitis B Antibodies, Hepatitis B Surface Antigens, Hepatitis B e Antigens, Hepatitis B virus, Hepatitis B, Chronic, Humans, Infant, Lamivudine, Male, Middle Aged, Polymerase Chain Reaction, Reproducibility of Results, Reverse Transcriptase Inhibitors, Sensitivity and Specificity, Viral Load, Journal Article",

author = "Mendy, {Maimuna E} and Steve Kaye and {van der Sande}, Marianne and Pura Rayco-Solon and Waight, {Pauline A} and Deborah Shipton and Dorka Awi and Paul Snell and Whittle, {Hilton C} and McConkey, {Samuel J}",

year = "2006",

month = apr,

day = "4",

doi = "10.1186/1743-422X-3-23",

language = "English",

volume = "3",

pages = "23",

journal = "Virology Journal",

issn = "1743-422X",

publisher = "BioMed Central",

}

TY - JOUR

T1 - Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia

AU - Mendy, Maimuna E

AU - Kaye, Steve

AU - van der Sande, Marianne

AU - Rayco-Solon, Pura

AU - Waight, Pauline A

AU - Shipton, Deborah

AU - Awi, Dorka

AU - Snell, Paul

AU - Whittle, Hilton C

AU - McConkey, Samuel J

PY - 2006/4/4

Y1 - 2006/4/4

N2 - BACKGROUND/AIM: The study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials.METHOD: Quantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers.RESULTS: The qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 x 103 to 1.5 x 108 copies/mL) and correlated well with those from a commercial assay (r = 0.91, (p < 0.001). Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% (161/209) of HBeAg negative and in all HBeAg positive carriers.CONCLUSION: This method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.

AB - BACKGROUND/AIM: The study aimed at developing a real-time quantitative PCR assay to monitor HBV serum virus load of chronic carriers enrolled in therapeutic trials.METHOD: Quantitative real-time PCR assay was carried out using SYBR-Green signal detection and primers specific to the S gene. Thermal cycling was performed in an ABi 5700 sequence detection system. The assay was calibrated against an international HBV DNA standard and inter- and intra-assay reproducibility determined. Levels of viral load were monitored for 1-year in lamivudine treated carriers. Correlation between HBV DNA levels and HBeAg sero-status was determined in untreated carriers.RESULTS: The qPCR assay showed good intra- and inter-assay reproducibility over a wide dynamic range (1.5 x 103 to 1.5 x 108 copies/mL) and correlated well with those from a commercial assay (r = 0.91, (p < 0.001). Viral load levels dropped dramatically but temporarily during and after a short course of lamivudine therapy. HBV DNA was a more reliable indicator of the presence of virus than HBe antigen and was detected in 77.0% (161/209) of HBeAg negative and in all HBeAg positive carriers.CONCLUSION: This method is reliable, accurate, and reproducible. HBV DNA Quantification by qPCR can be used to monitor the efficacy of HBV therapy and useful in understanding the natural history of HBV in an endemic area.

KW - Adolescent

KW - Adult

KW - Age Factors

KW - Aged

KW - Antiviral Agents

KW - Carrier State

KW - Child

KW - Child, Preschool

KW - DNA Primers

KW - DNA, Viral

KW - Gambia

KW - Hepatitis B Antibodies

KW - Hepatitis B Surface Antigens

KW - Hepatitis B e Antigens

KW - Hepatitis B virus

KW - Hepatitis B, Chronic

KW - Humans

KW - Infant

KW - Lamivudine

KW - Male

KW - Middle Aged

KW - Polymerase Chain Reaction

KW - Reproducibility of Results

KW - Reverse Transcriptase Inhibitors

KW - Sensitivity and Specificity

KW - Viral Load

KW - Journal Article

U2 - 10.1186/1743-422X-3-23

DO - 10.1186/1743-422X-3-23

M3 - Article

C2 - 16594999

SN - 1743-422X

VL - 3

SP - 23

JO - Virology Journal

JF - Virology Journal

ER -

Application of real-time PCR to quantify hepatitis B virus DNA in chronic carriers in The Gambia

Abstract

Keywords

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