VeriFiler™PlusPCR扩增试剂盒是一种6-dye多重测定,可同时扩增一组23个常染色体标记(D3S1358,vWA,D16S539,CSF1PO,D6S1043,D8S1179,D21S11,D18S51,D5S818,D2S441,D19S433,FGA,D10S1248,D22S1045,D1S1656,D13S317,D7S820,PentaE,PentaD,TH01、D12S391、D2S1338和TPOX),质量指标系统,和两个性别识别标记。合并,这些标记满足中国国家常染色体DNA数据库以及扩展的CODIS(组合DNA索引系统)的要求。VeriFilerPlus套件采用改进的MasterMix开发,它采用了更亮的TED™染料,并适应更高的样品加载量,从而提高灵敏度,并实现从具有挑战性的案例样本(包括触摸)中最大限度地恢复信息,退化,并抑制样品。这里,我们报告了遵循SWGDAM(DNA分析方法科学工作组)指南的发育验证研究的结果,其中包括基于PCR的研究数据,灵敏度,物种特异性,稳定性,精度,再现性和重复性,和谐,口吃,DNA混合物,和模拟案例样本的性能。结果验证了多路复用设计,并证明了套件的鲁棒性,可靠性,以及作为人类识别案例DNA样本的检测方法的适用性。
The VeriFiler™ Plus PCR Amplification Kit is a 6-dye multiplex assay that simultaneously amplifies a set of 23 autosomal markers (D3S1358, vWA, D16S539, CSF1PO, D6S1043, D8S1179, D21S11, D18S51, D5S818, D2S441, D19S433, FGA, D10S1248, D22S1045, D1S1656, D13S317, D7S820, Penta E, Penta D, TH01, D12S391, D2S1338, and TPOX), a quality indicator system, and two sex-identification markers. Combined, the markers satisfy the requirements of the Chinese National autosomal DNA database as well as expanded CODIS (Combined DNA Index System). The VeriFiler Plus kit was developed with an improved Master Mix which incorporates the brighter TED™ dye, and accommodates a higher sample loading volume thus allowing for increased sensitivity and enabling maximum information recovery from challenging casework samples including touch, degraded, and inhibited samples. Here, we report the results of the developmental validation study which followed the SWGDAM (Scientific Working Group on DNA Analysis Methods) guidelines and includes data for PCR-based studies, sensitivity, species specificity, stability, precision, reproducibility and repeatability, concordance, stutter, DNA mixtures, and performance on mock casework samples. The results validate the multiplex design as well as demonstrate the kit\'s robustness, reliability, and suitability as an assay for human identification with casework DNA samples.