HBV-DNA提取液I的配制和应用评价

Preparation of HBV-DNA Extract I and the Application Evaluation

目的探讨荧光定量PCR法检测HBV-DNA提取液I的配制并评价其应用效果。方法按照DNA提取原理及文献,筛选提取液I中有效成分PEG6000和Na Cl的最佳浓度,与原厂家核酸提取液I比对并评估其应用效果。结果在不同分子量PEG配制的提取液中,以PEG6000效果最好;PEG6000和Na Cl浓度分别为200g/L、250g/L组成的提取液I提取效果最佳;对30份不同浓度的HBV标本进行的检测结果显示,自配HBV-DNA提取液I与原厂家提取液无显著差异(P>0.05),两者相关性良好(r=0.96),变异系数分别为3.8%和3.4%。结论自配200g/L PEG6000和250g/L Na Cl浓度的HBV-DNA提取液I可用于HBV-DNA定量检测。

Objective To discuss the detection of the preparation of HBV-DNA extract I with fluorescent quantitative Polymerase Chain Reaction（FQ-PCR）and to evaluate the application effect. Methods According to DNA extracting mechanism and the literature,the best density of the effective ingredients PEG6000 and Na CL in the extract was screened out and matched with the nucleic acid extract of the manufacturer; the application effect was evaluated. Results Out of the extracts prepared in different molecular weight of PEG, PEG6000 was of best effect; the most effective extraction of extract I was gained when the density of PEG6000 and Na CL was at200g/L and 250g/L respectively; the detection of 30 HBV specimen of different density showed that the self-prepared HBV-DNA extract I was of no obvious difference from the extract of the manufacture.（P〉0.05）, the two was in good correlation（r=0.96）and the coefficients of variation were 3.8% and 3.4% respectively. Conclusions The self-prepared HBV-DNA extract I with PEG6000 at the density of 200g/L and Na CL at the density of 250g/L can be applied in the quantitative detection of HBV-DNA.