用引物Y_3,Y_4和DNA扩增技术作血液(痕)和毛根性别鉴定的研究

SEXING OF BLOOD(STAINS) AND HAIR ROOTS BY POLYMERASE CHAIN REACTION WITH PRIMERS Y3.Y4.

作者用引物Y_3、Y_4和DNA聚合酶链式反应(PCR)作微量人类血液(痕)和毛根的性别鉴定。扩增的靶序列位于Y染色体DNA特异3.4kb重复序列中,扩增产物为460bp。检材用量为:新鲜血液0.5μl、血痕纱纤维1mm、毛根单个。20例保存4个月的血痕与2例保存6年半的血痕性别判定结果均正确,无性别记载的保存9～11年的3例血痕显现了清晰的460bpY特异DNA扩增带。15例保存20天的自然脱落毛根性别判定结果均正确。本法省略了检材处理中的酚-氯仿抽提DNA等纯化步骤,既简化了实验操作,又减少了检验过程中外源DNA的污染机会和样品DNA的损耗,使这一性别鉴定方法更符合法医学实践的需要。

We have succeeded in doing sex identification of trace human blood(stains) and hair roots by polymerase chain reaction (PCR) with primers Y3, Y4. The amplified target sequences located in special 3.4kb repeat sequences of Y-chromosome DNA. The expected Y-chromosome special fragments were detected. the DNA sources were fresh blood 0.5μl, or dried Mood stain 1mm in size or single hair root. The detecting results of 20 samples of blood stains of four months old and 2 samples of blood stains of six and half a year old were correct. 460bp special Y DNA bands were determinated in 3 samples of unnknown blood stains kept for 9～11 years. The sex determination results of 15 samples of natual-shed hair roots were correct. The PCR method for the sex identification reported in this paper is easy and the DNA extraction procedure was omitted. It also reduce the chance of DNA contamination and loss of DNA specimens. Therefore, the sex determination by PCR method can meet the reqirement of the forensic practice.