一种用于降解DNA样本的性别鉴定方法

目的　建立一种采用PCR技术对降解DNA样本进行性别鉴定的新方法。　方法　采用针对amelogenin基因X染色体外显子 3bp缺失设计的引物AMELU1及AMELD1,对在室温环境下放置 5～ 15年的男、女血痕标本各 5 0例、毛发各 2 0例、骨骼各 2 0例以及现场提取 5 - - 2 0天的男、女腐败肌肉各 10例标本中提取的降解DNA样本进行扩增。用PAG( 9%T ,3 %C)电泳、银染显带检测扩增产物。　结果　所有样本均得到正确结果 ,男性检材表现为 83bp的Y特异性及 80bp的X特异性 2条谱带 ,而女性检材仅有 1条 80bp的X特异性谱带。　结论　用针对amelogenin基因X染色体外显子 3bp缺失设计的引物AMELU1及AMELD1鉴定性别的方法灵敏、可靠、方便 ,是降解DNA检材性别鉴定十分理想的方法。

A new method of Sex determination for degraded DNA samples X-gene.

Objective TO explore a new method of PCR-based sex test for degraded DNA samples. Methods Primers AMEL1 and AMEL2 were used to amplify degraded template DNA in 200 samples extracted from 50 samples of bloodstains, 20 samples of hair-roots and skeletal in each male and female remains from 5 to 15 years, as well as 10 sample of putrid muscle tissues in each male and female collecting from scene 5-20 days after death. PCR products were analyzed by denaturing polyacrylamide gel ( 9%T, 3%C ) and detected by silver-stain systems. Results Total 200 samples of degraded DNA were obtained ideally results. There were two specific PCR-fragments of 80 bp for the X-chromosome and 83 bp for the Y-chromosome in male samples, and only one specific PCR-fragment of 80 bp for the X-chromosome . Conclusion Primers AMEL1 and AMEL2 could used to determine the sex in samples of degraded template DNA. This method has some valuable features such as sensitive, reliable and easy to manage in common laboratory.