核酸提取方法对乙肝病毒定量PCR检测结果的影响

目的寻找简单、快速地提取血清乙型肝炎病毒(HBV)DNA并能应用于定量PCR测定的方法. 方法使用碱裂解法、酚氯仿法和煮沸裂解法同时提取血清HBV-DNA,比较荧光定量PCR测定的重复性和测定值的差异. 结果碱裂解法所得HBV-DNA量高于酚氯仿法和煮沸裂解法(P＜0.05),并且碱裂解法所得结果的重复性好于酚氯仿法和煮沸裂解法(CV分别为0.21,0.28和0.33). 结论碱裂解法提取HBV-DNA简单、快速,适用于HBV-DNA荧光定量测定.

The Influence of Nucleic Acid Extraction from Serum on HBV DNA Detection by Fluorescence Quantitative PCR

Objective To develop a simple and rapid method for extracting nucleic acid from serum on HBV-DNA detection by PCR. Methods an Alkaline lysis method was used to isolate serum HBV-DNA for fluorescence quantitative PCR and compared with conventional phenol-chloroform extraction method and boiling lysis method. Results The HBV-DNA quantitative values by PCR using the alkaline lysis were higher than the phenol-chloroform extraction method and boiling lysis method ( P<0.05). Moreover,The HBV-DNA quantitative values by PCR using the alkaline lysis were more reproducible than the phenol-chloroform extraction method and boiling lysis method (CV: 0.21,0.28 and 0.33,respectively). Conclusion The alkaline lysis method is simple,rapid and efficient for extraction of serum HBV-DNA and suitable for HBV-DNA quantitative PCR detection.