学科主题临床医学
Cellular chromosome DNA interferes with fluorescence quantitative real-time PCR detection of HBV DNA in culture medium
Pan, Xiao-Ben; Wei, Lai; Han, Jin-Chao; Gao, Yan
关键词hepatitis B virus interference quantification
刊名JOURNAL OF MEDICAL VIROLOGY
2008
DOI10.1002/jmv.21042
80期:1页:47-52
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Virology
研究领域[WOS]Virology
关键词[WOS]HEPATITIS-B-VIRUS ; QUANTIFICATION ; INFECTION
英文摘要

Fluorescence quantitative real-time PCR (FQ-PCR) is a recently developed technique increasingly used for clinical diagnosis by detection of hepatitis B virus (HBV) DNA in serum. FQ-PCR is also used in scientific research for detection of HBV DNA in cell culture. Understanding potential FQ-PCR interference factors can improve the accuracy of HBV DNA quantification in cell culture medium. HBV positive serum was diluted with culture medium to produce three test groups with HBV DNA levels of 5 x 107 copies/ml (high), 5 x 105 copies/ml (medium), and 5 x 103 copies/ml (low). Chromosome DNA was extracted from HepG2 cells and then added to high, medium, and low group samples at final concentrations of 0, 12.5, 25, 50, and 100 mu g/ml. The samples were quantified by FQ-PCR and data were evaluated using statistical software. No marked changes were seen in the quantitative curves for high level HBV DNA samples when the samples were supplemented with 0-100 mu g/ml of chromosome DNA. Interference was observed in medium level samples when 50 and 100 mu g/ml of chromosome DNA was added. Interference was also observed in low level HBV DNA samples when the concentration of added chromosome DNA was greater than 25 mu g/ml. The interference was eliminated when samples were digested by DNase I prior to PCR detection. In Conclusions, the presence of cellular chromosome DNA can interfere with the detection of HBV DNA by FQ-PCR. Removal of cellular chromosome DNA from culture media prior to FQ-PCR is necessary for reliable HBV DNA quantitative detection.

语种英语
WOS记录号WOS:000251335900006
引用统计
被引频次:4[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/64333
专题北京大学第二临床医学院_北京大学肝病研究所
作者单位Peking Univ Peoples Hosp, Peking Univ Hepatol Inst, Beijing 100044, Peoples R China
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GB/T 7714
Pan, Xiao-Ben,Wei, Lai,Han, Jin-Chao,et al. Cellular chromosome DNA interferes with fluorescence quantitative real-time PCR detection of HBV DNA in culture medium[J]. JOURNAL OF MEDICAL VIROLOGY,2008,80(1):47-52.
APA Pan, Xiao-Ben,Wei, Lai,Han, Jin-Chao,&Gao, Yan.(2008).Cellular chromosome DNA interferes with fluorescence quantitative real-time PCR detection of HBV DNA in culture medium.JOURNAL OF MEDICAL VIROLOGY,80(1),47-52.
MLA Pan, Xiao-Ben,et al."Cellular chromosome DNA interferes with fluorescence quantitative real-time PCR detection of HBV DNA in culture medium".JOURNAL OF MEDICAL VIROLOGY 80.1(2008):47-52.
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