DYS385检测方法的改进

目的建立检测DYS385的新方法。方法比较基因数据库(GDB)中推荐的引物和Schneider所设计的引物扩增DYS385的效果;在此基础上,以GDB推荐的引物作为外引物,Schneider所设计的引物作为内引物,通过调整内、外引物的浓度,优化扩增条件,建立DYS385的半巢式PCR体系。结果与常规方法相比,采用Schneider所设计的引物扩增DYS385,扩增片段缩短112bp,电泳分离的效果好,灵敏度提高2倍,达500pg;建立的半巢式扩增方法,能特异性扩增短片段,灵敏度提高20倍,达50pg。结论建立的DYS385半巢式扩增方法具有更高的特异性和灵敏度,适合法医学应用。

New improved approaches for DYS385 detection

Objective To establish more sensitive methods for detection of DYS385 in routine forensic casework. Methods The primers recommended by Genome Database (GDB) and Schneider were used to amplify DYS385 respectively. Then, a semi-nested PCR of DYS385 was designed by using the two different primers as outer and inner primer. A series of experiments were carried out to achieve good result by adjusting the ratio of outer/inner primer and optimizing the PCR condition. Results It showed that an overall 112bp shorter DYS385 fragments and better electrophoretic separation were obtained by using primer2B. By using the semi-nested PCR approach, the shorter specific DYS385 fragments could be amplified and detectable DNA amounted to 50pg. Conclusion This method is 20 fold more sensitive than the ordinary method.