8染料荧光标记法医Y染色体短串联重复序列(Y-STRs)试剂盒,62双工Y-STR多路放大系统,开发和优化。通过检测PCR条件对系统进行了验证,口吃比(SR)和峰高比,灵敏度,混合物样品,精度和准确度,物种特异性,和抑制研究根据科学工作组对DNA分析方法指南。基于PCR的研究表明,推荐的PCR条件已针对该试剂盒进行了优化。在敏感性研究中,从模板DNA获得完整的图谱,数量为u125pg。从三个不同的实验室获得一致的概况。所有基因座的SRs均小于15%,显示出良好的平衡和合适的平均峰高。在常见动物物种和微生物的谱中没有检测到峰。在男性-男性混合物研究中,所有基因座均以1:8的比例观察到,在男女混合研究中,如果男性DNA输入≥0.5ng/µL,则可以以1:500的比例对所有等位基因进行分析。抑制剂研究表明,该试剂盒对常见抑制剂的存在具有不同程度的抗性。人口研究表明,62-plexY-STR试剂盒提高了无关中国汉族男性(n=192)的歧视能力。当单倍型多样性为1时,62-plexY-STR试剂盒的鉴别力概率为0.9948,适用于法医调查。结果表明,所开发的8-染料荧光标记62位点系统是灵敏的,健壮,方便,和高度信息化的法医应用。
An 8-dye fluorescence-labeling forensic Y-chromosomal short tandem repeats (Y-STRs) kit, the 62-plex Y-STR multiplex amplification system, was developed and optimized. The system was validated by testing PCR conditions, stutter ratios (SR) and peak height ratios, sensitivity, mixture samples, precision and accuracy, species-specificity, and inhibition studies according to the Scientific Working Group on DNA Analysis Methods guidelines. PCR-based studies showed that the recommended PCR conditions were optimized for this kit. In the sensitivity study, a full profile was obtained from template DNA with a quantity of u125 pg. Consistent profiles were obtained from three different laboratories. The SRs in all loci were less than 15%, and nice balance and suitable average peak height were shown. No peaks were detected in the profiles of common animal species and microorganisms. In the male-male mixture studies, all loci were observed at a ratio of 1:8, and in the male-female mixture study, all alleles could be profiled at a ratio of 1:500 if the male DNA inputs were ≥0.5 ng/µL. An inhibitor study demonstrated that the kit had varying degrees of resistance to the presence of common inhibitors. Population study demonstrated the 62-plex Y-STR Kit improved the power of discrimination in unrelated Chinese Han males (n = 192). When haplotype diversity was 1, the probability of discrimination power of the 62-plex Y-STR Kit was 0.9948, which is suitable for forensic investigations. The results show that the developed 8-dye fluorescence labeling 62 loci system is sensitive, robust, convenient, and highly informative for forensic applications.