Developmental validation of the MGIEasy Signature Identification Library Prep Kit, an all-in-one multiplex system for forensic applications
Developmental validation of the MGIEasy Signature Identification Library Prep Kit, an all-in-one multiplex system for forensic applications
复制标题
MGIEasy 签名识别文库制备试剂盒的开发验证，这是一种用于法医应用的一体化多重系统
DOI：
10.1007/s00414-021-02507-0
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发表时间：
2021-02-01
影响因子：
2.1
通讯作者：
Sun, Hongyu
中科院分区：
文献类型：
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作者：
Li, Ran;Shen, Xuefeng;Sun, Hongyu
分析细胞核和线粒体基因组中的遗传标记有助于各种法医应用，例如个体识别和亲属关系分析。然而，大多数商业试剂盒分别检测这些标志物，这是耗时，费力，更容易出错（错误标记，污染，......）。MGIEasy Signature Identification Library Prep Kit（以下称为“MGIEasy鉴定系统”; MGI Tech，Shenzhen，China）已被设计成提供一种简单、快速和稳健的方法来在一个多重PCR反应中检测适当的标志物：52个常染色体STR，27个X染色体STR，48个Y染色体STR，145个身份信息SNP，53个祖先信息SNP，29个表型信息SNP，和线粒体DNA的高变区（mtDNA）。在此，我们根据DNA分析方法科学工作组（SWGDAM）的指导原则验证了MGIEasy鉴定系统的性能，评估了种属特异性、灵敏度、混合物鉴定、非最佳条件下的稳定性（降解样品、抑制剂污染和各种底物）、重复性和一致性。使用MGIEasy鉴定系统制备的文库在MGISEQ-2000仪器（MGI Tech）上测序。MGIEasy的STR、SNP和mtDNA基因型分别与CE的STR基因型（99.79%）、MiSeq FGX的SNP基因型（99.78%）和Sanger的mtDNA基因型（100%）高度一致。该系统具有很强的人类特异性，对四种常见的PCR抑制剂具有抗性，并且可靠地扩增了低量DNA（低至0.125 ng）和降解的DNA（~ 150 nt）。大多数来自次要贡献者的独特等位基因在1：10的男女混合物和男女混合物中检测到;一些次要Y-STR等位基因甚至在1：1000的男女混合物中检测到。MGIEasy还成功地直接扩增了FTA卡、滤纸和拭子上的血迹标记。因此，我们的研究结果表明，MGIEasy鉴定系统是适合用于法医分析，由于其强大的和可靠的性能，对不同的质量和数量的样品。
Analyzing genetic markers in nuclear and mitochondrial genomes is helpful in various forensic applications, such as individual identifications and kinship analyses. However, most commercial kits detect these markers separately, which is time-consuming, laborious, and more error-prone (mislabelling, contamination, ...). The MGIEasy Signature Identification Library Prep Kit (hereinafter “MGIEasy identification system”; MGI Tech, Shenzhen, China) has been designed to provide a simple, fast, and robust way to detect appropriate markers in one multiplex PCR reaction: 52 autosomal STRs, 27 X-chromosomal STRs, 48 Y-chromosomal STRs, 145 identity-informative SNPs, 53 ancestry-informative SNPs, 29 phenotype-informative SNPs, and the hypervariable regions of mitochondrial DNA (mtDNA). Here, we validated the performance of MGIEasy identification system following the guidelines of the Scientific Working Group on DNA Analysis Methods (SWGDAM), assessing species specificity, sensitivity, mixture identification, stability under non-optimal conditions (degraded samples, inhibitor contamination, and various substrates), repeatability, and concordance. Libraries prepared using MGIEasy identification system were sequenced on a MGISEQ-2000 instrument (MGI Tech). MGIEasy-derived STR, SNP, and mtDNA genotypes were highly concordant with CE-based STR genotypes (99.79%), MiSeq FGx-based SNP genotypes (99.78%), and Sanger-based mtDNA genotypes (100%), respectively. This system was strongly human-specific, resistant to four common PCR inhibitors, and reliably amplified both low quantities of DNA (as low as 0.125 ng) and degraded DNA (~ 150 nt). Most of the unique alleles from the minor contributor were detected in 1:10 male-female and male-male mixtures; some minor Y-STR alleles were even detected in 1:1000 male-female mixtures. MGIEasy also successfully directly amplified markers from blood stains on FTA cards, filter papers, and swabs. Thus, our results demonstrated that MGIEasy identification system was suitable for use in forensic analyses due to its robust and reliable performance on samples of varying quality and quantity.