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法医学杂志 2018, Vol. 34 Issue (4) :417-419,427    DOI: 10.12116/j.issn.1004-5619.2018.04.015
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尼龙膜套管分离技术对混合斑中精子细胞DNA的提取
马  骏1,童  奇2,高良弼1,朱  川1,江志强1
1. 长兴县公安司法鉴定中心,浙江 长兴 313100; 2. 安吉县公安司法鉴定中心,浙江 安吉 313300
Extraction of DNA from Sperm Cells in Mixed Stain by Nylon Membrane Bushing Separation Technique
MA Jun1, TONG Qi2, GAO Liang-bi1, ZHU Chuan1, JIANG Zhi-qiang1
1. Forensic Judicial Appraisal Center of Changxing Public Security Bureau, Changxing 313100, China; 2. Forensic Judicial Appraisal Center of Anji Public Security Bureau, Anji 313300, China

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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,AmpF?詛STR?誖Identifiler?誖Plus PCR扩增试剂盒进行PCR扩增,3500xL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。
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马骏
童奇
高良弼
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     
Abstract: 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?詛STR?誖Identifiler?誖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.
Keywordsforensic genetics   spermatid   membrane separation technology   mixed stain   casing, nylon membrane   differential pyrolysis     
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About author: 马骏(1981—),男,主要从事法医DNA技术研究和检验鉴定;E-mail:markbrianxy@163.com
引用本文:   
马骏,童奇,高良弼,等.尼龙膜套管分离技术对混合斑中精子细胞DNA的提取[J]  法医学杂志, 2018,34(4): 417-419,427
MA Jun, TONG Qi, GAO Liang-bi,et al..Extraction of DNA from Sperm Cells in Mixed Stain by Nylon Membrane Bushing Separation Technique[J]  法医学杂志, 2018,34(4): 417-419,427
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