血清中HBV-DNA的两种定量PCR方法的比较

Comparison of Two Different PCR Assays in the Quantitative Measurement of Serum Hepatitis B Virus DNA

目的比较乙型肝炎病毒脱氧核糖核酸(HBV-DNA)定量测定的两种方法:聚合酶链式反应-酶联免疫吸附测定(PCR-ELISA)法和荧光定量-聚合酶链式反应(FQ-PCR)法.方法检测了两种方法的灵敏度、批内批间可信度和特异性.结果PCR-ELISA法和FQ-PCR法在特异性和批内批间可信度方面相似.在85个HBV表面抗原阳性的病人血清中两种方法的检测结果96%保持一致,HBV-DNA血清阳性率分别为84%和80%,其结果取对数后有一定线性关系(r=0.588 9).PCR-ELISA法的灵敏度比FQ-PCR法高出一个稀释度,PCR-ELISA的最小检测限7.6×103拷贝/ml,而FQ-PCR为2.1×104拷贝/ml.但FQ-PCR法的技术复杂性要简单得多,分析时间(2 h)比PCR-ELISA(8 h)要短得多.结论两种方法各有优缺点,每个实验室都应根据各自的需求加以选择.

objective To compare two different PCR assays in the quantitative measurement of serum hepatitis B virus DNA: PCR-ELISA and fluorescence quantitative PCR (FQ-PCR). Methods Examined the sensitivity, linearity, variability and specificity of two methods. Results Results showed similar and satisfactory assay specificity,as well as mterassay and intra-assay variability values, for both PCR-ELISA and FQ-PCR. Ninty-six percent of 85 serum samples from hepatitis B surface antigen-positive patients showed concordant results with the two assays. The HBV-DNA seropositivity rates for the 85 samples were 84% and 80% by PCR-ELISA and FQ-PCR respectively .and a linear relationship (r = 0. 588 9)was observed between two sets of data after logarithmic conversion. The PCR-ELTSA was little more sensitive than the FQ-PCR assay by 1 dilution,with the lowest reading being 7. 6×103 copy/ml (2.1 × 104copy/ml by FQ-PCR). But FQ-PCR assay was technically much less complex and required a much shorter assay time (2 h) than the PCR-ELISA(8 h). Conclusions The two methods have their advantages and disadvantages,should make a suitable choice by own needs.