应用SNaPshot技术检测精液特异性cSNP遗传标记  

Application of SNaPshot Technology in Semen-Specific cSNP Genetic Marker

作　　者：陶瑞旸 王守宇 袁春艳 夏若成 李成涛 TAO Rui-yang;WANG Shou-yu;YUAN Chun-yan;XIA Ruo-cheng;LI Cheng-tao(Shanghai Key Laboratory of Forensic Medicine,Key Laboratory of Forensic Science,Ministry of Justice,Shanghai Forensic Service Platform,Academy of Forensic Science,Shanghai 200063,China;Department of Forensic Medicine,School of Basic Medical Sciences,Fudan University,Shanghai 200032,China;Department of Forensic Medicine,Inner Mongolia Medical University,Hohhot 010110,China)

机构地区：[1]司法鉴定科学研究院、上海市法医学重点实验室、司法部司法鉴定重点实验室、上海市司法鉴定专业技术服务平台,上海200063 [2]复旦大学基础医学院法医学系,上海200032 [3]内蒙古医科大学法医学教研室,内蒙古呼和浩特010030

出　　处：《法医学杂志》2023年第5期465-470,共6页Journal of Forensic Medicine

基　　金：国家自然科学基金资助项目(82202080,81930056);中青年科技创新领军人才资助项目(2018RA2102);上海市法医学重点实验室资助项目(21DZ2270800);司法部司法鉴定重点实验室资助项目;上海市司法鉴定专业技术服务平台资助项目。

摘　　要：目的探讨基于SNaPshot技术的精液特异性编码区单核苷酸多态性(coding region single nucleo⁃tide polymorphism,cSNP)遗传标记检测在精液(斑)溯源及混合体液(斑)鉴定中的可行性。方法制备16例精液斑和11例精液-静脉血混合斑样本,提取其基因组DNA(genomic DNA,gDNA)和总RNA,并将总RNA逆转录为互补DNA(complementary DNA,cDNA)。在已验证的精液特异性mRNA编码基因上筛选cSNP遗传标记。基于SNaPshot技术构建cSNP复合检测体系并通过CE对样本进行基因分型。结果成功构建了包含5个精液特异性cSNP的复合检测体系。在16例精液样本中,除了位于TGM4基因上的cSNP在cDNA的检测结果中出现等位基因丢失外,其余cSNP的gDNA和cDNA分型结果高度一致。检测精液-静脉血混合斑时,检出的cSNP分型结果均与精液提供者的基因型一致,未受到静脉血提供者基因型的干扰。结论应用SNaPshot技术检测精液特异性cSNP的方法可以应用于法医学精液(斑)的基因分型,并为混合体液(斑)中精液来源个体的判定提供信息。Objective To explore the feasibility of genetic marker detection of semen-specific coding region single nucleotide polymorphism(cSNP)based on SNaPshot technology in semen stains and mixed body fluid identification.Methods Genomic DNA(gDNA)and total RNA were extracted from 16 semen stains and 11 mixtures composed of semen and venous blood,and the total RNA was reverse transcribed into complementary DNA(cDNA).The cSNP genetic markers were screened on the validated semen-specific mRNA coding genes.The cSNP multiplex detection system based on SNaP-shot technology was established,and samples were genotyped by capillary electrophoresis(CE).Results A multiplex detection system containing 5 semen-specific cSNPs was successfully established.In 16 semen samples,except the cSNP located in the TGM4 gene showed allele loss in cDNA detection results,the gDNA and cDNA typing results of other cSNPs were highly consistent.When detecting semen-venous blood mixtures,the results of cSNP typing detected were consistent with the genotype of semen donor and were not interfered by the genotype of venous blood donor.Conclusion The method of semen-specific cSNPs detection by SNaPshot technology method can be applied to the genotyping of semen(stains)and provide information for determining the origin of semen in mixed body fluids(stains).

关 键 词：法医遗传学 体液鉴定 混合斑鉴定 编码区单核苷酸多态性 SNapShot技术 精液 

分 类 号：R89[医药卫生—法医学] DF795.2[医药卫生—临床医学] D919.2[政治法律—诉讼法学]