Objectives: 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.
Conclusions: 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.
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。.
Keywords: Felis catus; forensic genetics; individual identification; multiplex amplification system; sex determining region of Y (SRY); short tandem repeat.