Abstract:
The idea of developing a new multiplex STR amplification system was conceived in 2011 as an effective way to implement the new European standard set (ESS) of 12 STR markers adopted by The Council of the European Union in 2009 while maintaining an effective compatibility and information exchange with the historical DNA profiles contained in the Spanish national DNA database (around 200,000 DNA profiles) mainly based on the 13 CODIS core STR loci plus D19S433 and D2S1338 markers. With this goal in mind we proposed to test and validate a single STR amplification system for simultaneous analysis of 21 STR markers covering both CODIS and ESS core STR loci plus three additional markers (D19S433, D2S1338, and SE33) also contained in commonly used STR kits and national DNA databases. In 2012, we started the first beta-testing with a 6-dye STR kit prototype containing 24 loci (now known as the GlobalFiler™ PCR Amplification Kit) developed by Life Technologies in response to the CODIS Core Loci Working Group\'s recommendation to expand the CODIS Core Loci. This prototype included our proposal of 21 autosomal STR markers and two Y-chromosome markers (DYS391 and Y-indel) and maximizes concordance with established databases and previously analyzed samples by maintaining primer sequences of previous Identifiler(®)/NGM SElect™ kits for the 21 STR markers except for TPOX. This paper describes the validation studies conducted with the first commercial available 6-dye STR kit for casework using a 3500 genetic analyzer for fragment detection that included the analysis of the following parameters and aspects: analytical threshold, sensitivity & stochastic threshold, heterozygous balance, stutter threshold, precision and accuracy, repeatability and reproducibility, genotype concordance, DNA mixtures, species specificity, and stability studies with case type samples. The studies demonstrated that the GlobalFiler™ system provided equivalent overall performance to previous forensic STR PCR kits, but with enhanced discrimination power for a better match efficiency that would reduce the chance of adventitious matches during DNA data exchange among national DNA databases.
摘要:
2011年提出了开发新的多重STR扩增系统的想法,作为实施欧盟理事会于2009年通过的12个STR标记的新欧洲标准集(ESS)的有效方法,同时保持有效的兼容性和信息交换与西班牙国家DNA数据库中包含的历史DNA图谱(约200,000个DNA图谱)主要基于13个CODIS核心STR基因座加上D19S433和D2S1338。考虑到这个目标,我们建议测试和验证单个STR扩增系统,以同时分析涵盖CODIS和ESS核心STR基因座的21个STR标记以及常用STR试剂盒和国家DNA数据库中包含的三个其他标记(D19S433,D2S1338和SE33)。2012年,我们开始使用包含24个基因座的6染料STR试剂盒原型(现在称为GlobalFiler™PCR扩增试剂盒)进行首次beta测试,该试剂盒由LifeTechnologies开发,以响应CODIS核心基因座工作组的建议。该原型包括我们提出的21个常染色体STR标记和两个Y染色体标记(DYS391和Y-indel)的建议,并通过保留先前的Identifier(®)/NGMSElect™试剂盒的引物序列,最大程度地与已建立的数据库和先前分析的样品保持一致除了TPOX之外的21个STR标记。本文介绍了第一个商业可用的6-dyeSTR试剂盒进行的案例工作的验证研究,使用3500基因分析仪进行片段检测,包括以下参数和方面的分析:分析阈值,灵敏度和随机阈值,杂合平衡,口吃阈值,精度和准确度,重复性和再现性,基因型一致性,DNA混合物,物种特异性,以及案例类型样本的稳定性研究。研究表明,GlobalFiler™系统提供了与以前的法医STRPCR试剂盒相当的整体性能,但是具有增强的辨别能力以获得更好的匹配效率,这将减少在国家DNA数据库之间进行DNA数据交换期间偶然匹配的机会。