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