固相载体法提取病毒性乙型肝炎患者血清DNA的研究

A Solid Phase Method for Serum HBV DNA Extraction

【目的】寻找简单快速地提取血清乙型肝炎病毒(HBV)DNA的方法。【方法】分别用盐酸胍-硅胶法、酚氯仿法和煮沸裂解法同时提取血清HBV DNA并应用于荧光定量PCR。【结果】盐酸胍-硅胶法所得HBV DNA量高于酚氯仿法和煮沸裂解法(P<0.05),并且盐酸胍-硅胶法所得结果的重复性优于酚氯仿法和煮沸裂解法(CV分别为0.28,0.31和0.37)。【结论】盐酸胍-硅胶法提取HBV DNA简单、快速且可靠,适用于HBV DNA定量测定。

[Objective]To develop a simple and rapid method for extracting HBV DNA from serum. [Methods]GuHCl-silica 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 GuHCl-silica were higher than those using phenol-chloroform extraction method and boiling lysis method（ P d0.05）. Morever, the HBV DNA quantitative values by PCR using the GuHCl-silica were more reproducible than those using phenol-chloroform method and boiling lysis method（CV：0. 28,0. 31 and0.37 ,respectively）. [Conclusion]The GuHCl-silica method is simple, rapid and efficient method for extraction of serum HBV DNA and suitable for HBV DNA quantitative PCR detection.