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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。
Objective To establish a new method for sex identification of degraded DNA samples by PCR. Methods The primers AMELU1 and AMELD1 designed for the deletion of exon 3 of amelogenin gene were used in the study. Fifty cases of male and female blood stained specimens were collected at room temperature for 5 to 15 years. Twenty (20) cases of hair and 20 Cases, as well as on-site extraction of degraded DNA samples extracted from 10 specimens from 5 to 20 days of male and female spoiled muscles. PAG (9% T, 3% C) electrophoresis, silver staining bands detected amplification products. Results The correct results were obtained in all the samples. The male samples showed 83bp Y-specific and 80bp X-specific bands, while the female samples had only one 80bp X-specific band. Conclusion The method of identifying the sexes by primers AMELU1 and AMELD1 designed for the deletion of exon 3 of amelogenin gene is sensitive, reliable and convenient. It is a very ideal method to degrade DNA samples.