摘要
目的建立新型的混合斑中精子细胞分离的方法,并评价其应用价值。方法收集性侵案件中的40份混合斑检材,分别采用常规差异裂解法和尼龙膜套管分离技术进行精子细胞分离,使用硅膜试剂盒(Forensic DNA Extraction Kit for Soft Tissues)提取精子细胞DNA,Amp FlSTR~ Identifiler~ Plus PCR扩增试剂盒进行PCR扩增,3500x L基因分析仪进行电泳检测,并对两种分离方法的结果进行对比。结果 40份混合斑检材中,采用尼龙膜套管分离技术的检材仅3份有女性成分微弱残留,余均获得了完整的单一男性分型。而采用常规差异裂解法分离的检材中,有25份完全未检出男性精子细胞STR分型,15份检出男性精子细胞STR分型(7份不完整男性精子细胞STR分型,6份有女性成分残留,2份单一的男性精子细胞STR分型)。结论本研究建立的尼龙膜套管分离技术适用于混合斑中精子细胞的分离,特别是对含有大量女性细胞而精子细胞较少的检材提取效果明显,整个提取实验成本低廉、快速简便。
Objective To establish a novel method for the separation of sperm cells in mixed stain,and to evaluate its application value.Methods Totally 40 mixed stain samples were collected from sexual assault cases.Sperm cells were separated by the conventional differential lysis method and the nylon membrane bushing separation technique,respectively.The DNA of sperm cells was extracted with the silicon membrane kit(Forensic DNA Extraction Kit for Soft Tissues).The PCR amplification was performed using AmpF?STR?Identifiler?Plus kit,and the products were electrophoresed by 3500xL genetic analyser.The results of two separation methods were then compared.Results Complete and single-source male STR genotypes could be obtained from all the 40 mixed stain samples except three samples with minimal residual of female DNA by the nylon membrane bushing separation technique.The STR genotypes of sperm cells could not be detected in 25 samples,which were obtained in 15 samples(seven were of incomplete male STR genotypes,six with residual of female DNA,two were complete and single-source STR genotypes of sperm cells).Conclusion The nylon membrane bushing separation technique developed in present study can be used in the separation of sperm cells in mixed stain,especially for the extraction of a small amount of sperm from a large quantity of female cells,which is inexpensive,rapid and simple.
作者
马骏
童奇
高良弼
朱川
江志强
MA Jun;TONG Qi;GAO Liang-bi;ZHU Chuan;JIANG Zhi-qiang(Forensic Judicial Appraisal Center of Changxing Public Security Bureau,Changxing 313100,China;Forensic Judicial Appraisal Center of Anji Public Security Bureau,Anji 313300,China)
出处
《法医学杂志》
CAS
CSCD
2018年第4期417-419,427,共4页
Journal of Forensic Medicine
关键词
法医遗传学
精子细胞
膜分离技术
混合斑
尼龙膜套管
差异裂解
forensic genetics
spermatid
membrane separation technology
mixed stain
casing,nylon membrane
differential pyrolysis
