Institute of Forensic Medicine, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, 610041, PR China; Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Academy of Forensic Sciences, Ministry of Justice, Shanghai, 200063, PR China.
Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Academy of Forensic Sciences, Ministry of Justice, Shanghai, 200063, PR China; Department of Forensic Science, Medical School of Soochow University, Suzhou, 215123, PR China.
Forensic Sci Int Genet. 2019 Jul;41:128-136. doi: 10.1016/j.fsigen.2019.04.008. Epub 2019 Apr 30.
In addition to commonly used short tandem repeats (STRs) and single nucleotide polymorphisms (SNPs), insertion and deletion polymorphisms (InDels) have considerable potential in the field of forensic genetics because they combine desirable characteristics of both STRs and SNPs. In the present study, the SifaInDel 45plex system was designed to amplify 45 InDel markers, including 27 autosomal InDels (A-InDels), 16 X chromosome InDels (X-InDels) and two Y chromosome InDels (Y-InDels), simultaneously in a single PCR procedure and then detect products by capillary electrophoresis (CE). We also optimized the PCR conditions for the novel panel and performed several validation studies including repeatability/reproducibility, concordance, accuracy, sensitivity, stability, species specificity and population genetics. The results confirmed that full profiles could be obtained from ≥62.5 pg of input DNA and from a series of challenging samples encountered in routine casework. The SifaInDel 45plex panel could tolerate different concentrations of inhibitors, such as ≤50 μM hematin, ≤20 ng/μL nigrosine and ≤8000 ng/μL urea. In a population investigation, for the 27 A-InDels, the combined power of discrimination (CPD) exceeded 0.999999, and the combined power of exclusion in duos (CPE) and trios (CPE) was 0.955118 and 0.997754, respectively. For the 16 X-InDels, the combined PD and PD was computed as 0.999845 and 0.999998, respectively, and the combined mean exclusion chance in father/daughter or mother/son duos (MEC) and mean exclusion chance in standard trios involving daughters (MEC) was 0.976220 and 0.998163, respectively. In addition, the two Y-InDels could play a role in correctly determining gender. Overall, the established SifaInDel 45plex panel is a well-performing, reliable and robust multiplex system that stands out for combining a considerable number of A-indels, X-indels and Y-indels based on a CE platform. The population study results also demonstrated that the SifaInDel 45plex panel could be a valid complementary approach for human identification and complex kinship analysis.
除了常用的短串联重复序列（STRs）和单核苷酸多态性（SNPs）外，插入和缺失多态性（InDels）在法医学遗传学领域具有相当大的潜力，因为它们结合了 STRs 和 SNPs 的理想特征。在本研究中，设计了 SifaInDel 45plex 系统，以在单个 PCR 程序中同时扩增 45 个 InDel 标记物，包括 27 个常染色体 InDels（A-InDels）、16 个 X 染色体 InDels（X-InDels）和两个 Y 染色体 InDels（Y-InDels），然后通过毛细管电泳（CE）检测产物。我们还优化了该新型试剂盒的 PCR 条件，并进行了多项验证研究，包括重复性/可再现性、一致性、准确性、灵敏度、稳定性、种属特异性和群体遗传学。结果证实，从≥62.5pg 的输入 DNA 和在常规案例工作中遇到的一系列有挑战性的样本中，都可以获得完整的图谱。SifaInDel 45plex 试剂盒可以耐受不同浓度的抑制剂，如≤50μM 正铁血红素、≤20ng/μL 硝嗪和≤8000ng/μL 尿素。在群体研究中，对于 27 个 A-InDels，组合鉴别力（CPD）超过 0.999999，双亲和三人组的组合排除力（CPE）分别为 0.955118 和 0.997754。对于 16 个 X-InDels，组合 PD 和 PD 分别计算为 0.999845 和 0.999998，涉及父女或母子双亲和包含女儿的标准三人组的平均排除几率（MEC）分别为 0.976220 和 0.998163。此外，两个 Y-InDels 可以在正确确定性别方面发挥作用。总体而言，建立的 SifaInDel 45plex 试剂盒是一种性能良好、可靠且强大的多重系统，它基于 CE 平台，结合了相当数量的 A-InDels、X-InDels 和 Y-InDels。群体研究结果还表明，SifaInDel 45plex 试剂盒可以作为人类识别和复杂亲属关系分析的有效补充方法。
