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首页> 外文期刊>Journal of Clinical Microbiology >A Genotype-Independent Real-Time PCR Assay for Quantification of Hepatitis B Virus DNA
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A Genotype-Independent Real-Time PCR Assay for Quantification of Hepatitis B Virus DNA

机译：非基因型实时荧光定量PCR检测乙型肝炎病毒DNA

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Accurate quantification of hepatitis B virus (HBV) DNA levels is important for monitoring patients with chronic HBV infection and for assessing their responses to antiviral therapy. This study aimed to develop a real-time PCR assay that is sensitive and can accurately quantify a wide range of HBV DNA levels across the known HBV genotypes. An “in-house” real-time PCR assay using primers and a TaqMan probe in a highly conserved region of the HBV surface gene was designed. The assay was standardized against a WHO standard and validated against plasmids of HBV genotypes A through H. The linear quantification range was approximately 5 × 100 to 2.0 × 109 IU/ml. Results of samples from patients infected with HBV genotypes A through H tested using our real-time “in-house” PCR assay showed an excellent correlation with those of the Cobas Amplicor HBV Monitor (R2 = 0.9435) and the Cobas TaqMan HBV (R2 = 0.9873) tests. We have established a real-time PCR assay that is genotype independent and can accurately quantify a wide range of HBV DNA levels. Further studies of additional samples are ongoing to validate the genotype independence of our assay.
机译：准确定量乙肝病毒（HBV）DNA水平对于监测慢性HBV感染患者和评估其对抗病毒治疗的反应非常重要。这项研究旨在开发一种实时PCR检测方法，该检测方法灵敏并且可以准确量化已知HBV基因型中广泛的HBV DNA水平。设计了使用引物和TaqMan探针在HBV表面基因高度保守的区域内进行的“内部”实时PCR分析。该测定方法根据WHO标准进行了标准化，并针对A至H基因型HBV质粒进行了验证。线性定量范围约为5×10 0 至2.0×10 9 IU /毫升使用我们的实时“内部” PCR试验检测了感染了A型至H型HBV基因的患者的样品，结果与Cobas Amplicor HBV Monitor（ R 2 = 0.9435）和Cobas TaqMan HBV（ R 2 = 0.9873）测试。我们已经建立了一种实时PCR检测方法，该检测方法与基因型无关，可以准确定量各种HBV DNA水平。其他样品的进一步研究正在进行中，以验证我们测定的基因型独立性。

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