猫STR基因座复合扩增体系的构建及其法医学应用  

Establishment of Multiplex Amplification System of STR Loci in Felis Catus and Its Forensic Application

作　　者：席世涵 屈轶龄 夏若成 熊磊 柴思雨 佟春兰 陶瑞旸 李成涛 XI Shi-han;QU Yi-ling;XIA Ruo-cheng;XIONG Lei;CHAI Si-yu;TONG Chun-lan;TAO Rui-yang;LI Cheng-tao(Clinical Medical School of Inner Mongolia Minzu University,Tongliao 028000,Inner Mongolia,China;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,Medical College of Soochow University,Suzhou 215123,Jiangsu Province,China;De-partment of Forensic Medicine,School of Basic Medicine,Inner Mongolia Medical University,Hohhot 010030,China;School of Preclinical Medicine of Zunyi Medical University,Guizhou 563000,Zunyi Province,China)

机构地区：[1]内蒙古民族大学临床医学院,内蒙古通辽028000 [2]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 [3]苏州大学医学部法医学系,江苏苏州215123 [4]内蒙古医科大学基础医学院法医学系,内蒙古呼和浩特010030 [5]遵义医科大学基础医学院,贵州遵义563000

出　　处：《法医学杂志》2022年第2期231-238,245,共9页Journal of Forensic Medicine

基　　金：国家自然科学基金资助项目(81930056);中青年科技创新领军人才项目(2018RA2102);上海市法医学重点实验室资助项目(21DZ2270800);上海市司法鉴定专业技术服务平台资助项目(19DZ2292700);司法部司法鉴定重点实验室资助项目。

摘　　要：目的构建一个猫STR基因座复合扩增体系并对其技术性能进行测试,评估其应用价值。方法整理和分析猫STR基因座文献数据资料,筛选可用于猫个体识别和亲缘关系鉴定的STR基因座和性别鉴定位点,设计荧光标记引物,构建复合扩增体系。对构建的复合扩增体系进行灵敏度、准确性、均衡性、稳定性、种属特异性、组织同一性和混合样本等验证,并对145例猫无关个体进行群体遗传学调查。结果成功筛选到16个猫常染色体STR基因座和1个Y染色体性别决定区,并构建了包含上述基因座的复合扩增体系。DNA模板量低至0.25 ng时仍可得到完整分型,检测其他常见动物样本时未见特异性扩增峰。群体遗传学调查结果显示,16个猫STR基因座累积个体识别率为1-3.57×10^(-20),累积非父排除率为1-6.35×10^(-5),累积匹配概率为3.61×10^(-20)。结论本研究构建的猫STR基因座复合扩增体系灵敏度高、种属特异性好、分型结果准确,能够为司法鉴定领域中涉及猫的种属鉴定、个体识别及亲缘关系鉴定等案件提供有效的解决方法。Objective To construct a Felis catus STR loci multiplex amplification system and to evaluate its application value by testing the technical performance.Methods The published Felis catus STR loci data were reviewed and analyzed to select the STR loci and sex identification loci that could be used for Felis catus individual identification and genetic identification.The fluorescent labeling primers were designed to construct the multiplex amplification system.The system was validated for sensitivity,accuracy,balance,stability,species specificity,tissue identity and mixture analysis,and investigated the genetic polymorphisms in 145 unrelated Felis catus samples.Results Sixteen Felis catus autosomal STR loci and one sex determining region of Y(SRY)were successfully selected,and constructed a multiplex amplification system containing the above loci.The complete profile of all alleles could still be obtained when the amount of DNA template was as low as 0.25 ng.There was no specific amplification peak in other common animal samples.Population genetic surveys showed that total discrimination power(TDP)of the 16 STR loci was 1-3.57×10^(-20),the cumulative probability of exclusion(CPE)was 1-6.35×10^(-5) and the cumulative probability of matching was 3.61×10^(-20).Conclusion The Felis catus STR multiplex amplification system constructed in this study is highly sensitive,species-specific,and accurate in typing results,which can provide an effective solution for Felis catus species identification,individual identification and kinship identification in the field of forensic science.

关 键 词：法医遗传学 短串联重复序列 Y染色体性别决定区 复合扩增体系 个体识别 猫 

分 类 号：DF795.2[医药卫生—法医学]