建立一种HBV突变检测的蝎子实时荧光PCR新方法

目的：为 HBV 突变检测建立一种有效、快速、简捷的蝎子实时荧光 PCR 新方法。方法同时用蝎子实时荧光 PCR 新方法和 Sanger 测序法对比：对157例 HBV 患者的血清的 DNA 进行 HBV 基因rtN236T 突变检测，统计其符合率；通过对混有不同比例的 HBV 基因 rtN236T 突变核酸的样本进行对照检测来比较两者的灵敏度。结果在157例 HBV 患者的血清样本中，蝎子实时荧光PCR 新方法检测到 rtN236T 突变的有69例，而 Sanger 测序法检测的有68例，其符合率为98.6％；蝎子实时荧光PCR 新方法能检测出混有5％突变的样本，其灵敏度达5％，比 Sanger 测序法高。结论蝎子实时荧光 PCR 新方法能有效检测到 HBV 突变，且比 Sanger 测序法更为简单、快速、灵敏度高，适合临床应用。

Establishment of a new Scorpion real-time fluorescence PCR method to detect the mutation of HBV

Objective To establish an effective, rapid and simple Scorpion real-time PCR method to detect the mutation of HBV. Methods The rtN236T mutation of HBV was detected by Scorpion real-time PCR and Sanger sequencing method, respectively. 157 patients were tested and coincidence rate was calculated. The samples which mixed with different proportions of HBV mutant DNA were tested to compare the sensitivity of Scorpion real-time PCR method and Sanger sequencing method. Results In the DNA of 157 HBV patients, 69 cases with the rtN236 mutation of HBV were detected by Scorpion real-time PCR method while 68 cases were detected by Sanger sequencing method. The coincidence rate of the two methods was 98.6%. The Scorpion real-time PCR method can detect the samples which mixed with 5%HBV mutant DNA , and its sensitivity was 5%, which was higher than that of Sanger sequencing method. Conclusion The mutation of HBV can be detected by the Scorpion real-time PCR. It is more simple, convenient, high sensitivity than Sanger sequencing method. Scorpion real-time PCR method is suitable for clinical applications.