裂解液加热煮沸法在血清HBV DNA抽提中的应用  

The Application of Lysis-boiling Method in Extracting HBV DNA from Serum Sample

作　　者：刘晓颖[1] 马敏[2] 伍晓菲[1] 郑岚[1] 贾尧[1] 王迅[1] 

机构地区：[1]上海市血液中心,200051 [2]华东师范大学

出　　处：《临床输血与检验》2007年第3期234-236,共3页Journal of Clinical Transfusion and Laboratory Medicine

摘　　要：目的建立裂解液加热煮沸法抽提血液HBV DNA的方法。方法选取浓度分别为106、105、104、103copies/ml的HBV DNA阳性血清,采用5种方法抽提HBV DNA,并用SYBR GreenI荧光定量PCR检测抽提产物。结果在HBV DNA浓度为106、105、104copies/ml时5种抽提法(异硫氰酸胍加热煮沸/盐酸胍加热煮沸/kit/SDS加热煮沸/chelex-100加热煮沸)均能得到阳性结果,而在HBV DNA为低浓度103copies/ml时,只有异硫氰酸胍加热煮沸/盐酸胍加热煮沸/chelex-100加热煮沸能得到阳性结果。结论chelex-100裂解液加热煮沸抽提HBV DNA操作简便、省时、经济,为血液标本HBV基因检测在临床上的大规模应用奠定基础。Objective To establish new method to extract HBV DNA from serum sample. Methods Three serum samples with different HBV concentration （10^6,10^5,10^4, 10^3 copies/ml） were prepared with five methods for isolation of HBV DNA. HBV DNA template was detected by using SYBR Green Ⅰ real-time PCR. Results Five methods of template preparation （guanidine thiocyanate-boiling/ guanidine hydrochloride-boiling/ kit/ SDS-boiling/ chelex-100-boiling） were shown to be positive for HBV DNA amplicon at the concentration of 10^6,10^5,10^4 copies/ml after SYBR Green Ⅰ real-time PCR. While these methods were shown to be positive for HBV DNA at the concentration of 10^3 copies/ml. Conclusion The chelex-100-boiling methods to extract HBV DNA from serum sample is convenient, less time and consuming, and may be applied to large scale of clinical detection of HBV.

关 键 词：血清 乙型肝炎病毒 DNA裂解液煮沸抽提 

分 类 号：R512.62[医药卫生—内科学] R446.11[医药卫生—临床医学]