目的:在案发现场快速准确的锁定犯罪嫌疑人性别,缩小嫌疑人范围,为基层公安机关案件侦查提供技术支持.方法:通过对碱裂解法进行改良并建立人类凝固血液的基因组DNA提取方法;根据Y染色体的SRY基因序列设计特异性引物,优化PCR扩增反应条件,扩增产物经SYBR Green I染色后紫外光下观察.结果:改良碱裂解法中组②所得DNA浓度为(271.08±143.28) ng/μL,显著优于组①所得(P＜0.05),且经ITS序列验证均能扩增出目的条带;在PCR扩增产物加入SYBR Green Ⅰ染料后于紫外光下观察,男性血样的PCR扩增产物呈现绿色荧光,而女性血样的PCR扩增产物未有荧光.结论:使用改良碱裂解法中的组②快速提取案发现场凝血DNA,进行特异性PCR扩增后,观察加入SYBR Green Ⅰ后的PCR产物紫外光下是否有绿色荧光即能判断性别,该方法准确、可靠,较常规方法缩短1～2 h.
A Study on the Methodology of Rapid Identification on Sex by SRY Gene in Human Blood
Objective:In order to target the gender of the suspect at the scene quickly and accurately,narrow down the range of the suspect,and provide technical support for basic level public security organs in case investigation.Methods:Improved the method of alkaline lysis and established the method of extracting genomic DNA from human clotting blood.Designed specific primers according to the SRY gene sequence of Y chromosome,and optimized PCR amplification conditions of reaction.Observed the amplified products stained with SYBR Green Ⅰ under UV light.Results:The concentration of DNA derived from Group Ⅱ of modified alkaline lysis method was (271.08 ± 143.28) ng/μL,which was significantly better than that in group Ⅰ (P ＜ 0.05).The ITS sequence validation could amplify the target bands.Observe PCR amplification products added SYBR Green Ⅰ dye under UV light,green fluorescence could be observed in male blood samples,while it could not be observed in female blood samples.Conclusion:Compared with the traditional way,this method can shorten 1-2 hours in identifying the information of gender.And it is accurate and reliable.Therefore,it can narrow down the range of the suspect,and provide reference for the basic level public security organs in case investiong.
rapid identification of gendertwo-temperature PCR methodSRYblood coagulation
NETL
NSTL
万方数据
维普
