Forensic validation of the SNPforID 52-plex assay
Forensic validation of the SNPforID 52-plex assay
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DOI：
10.1016/j.fsigen.2007.01.004
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发表时间：
2007-06-01
影响因子：
3.1
通讯作者：
Court, Denise Syndercombe
中科院分区：
文献类型：
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作者：
Musgrave-Brown, Esther;Ballard, David;Court, Denise Syndercombe
单核苷酸多态性（SNP）分型在法医遗传学中的优势是众所周知的，包括更广泛的高通量分型平台的选择，较低的突变率，以及改善降解样品的分析。然而，如果SNPs要成为目前短串联重复序列（STR）分型方法的现实补充，它们必须被证明能够成功可靠地分析在个案工作情况下经常遇到的具有挑战性的样本。欧洲SNPforID联盟，由欧盟增长计划支持，已经开发了一种用于法医分析的52个SNPs的多重，在单个反应中扩增所有52个基因座，然后进行两个单碱基延伸（SBE）反应，用毛细管电泳检测。为了验证该测定，选择多种DNA提取物来代表法医案例工作中常见的问题，例如低拷贝数和降解。本研究共使用了40种浸提液。每份样本会送往五间参与实验室中的两间，以一式两份或三份的形式进行分型。实验室被指示进行分析，就像他们处理正常的个案样本一样。将结果报告给协调实验室，并与使用Powerplex（R）16（Promega）从相同提取物的传统STR分型获得的结果进行比较。这些结果表明，尽管成功分型高质量、低拷贝数提取物的能力较低，但52重SNP测定法在降解样本以及降解且数量有限的样本上的表现优于STR分型，这表明SNP分析可以在法医相关情况下提供优于STR分析的优势。然而，也有额外的问题所产生的污染和引物质量问题，这些进行了讨论。(c)2007爱思唯尔爱尔兰有限公司保留所有权利。
The advantages of single nucleotide polymorphism (SNP) typing in forensic genetics are well known and include a wider choice of high-throughput typing platforms, lower mutation rates, and improved analysis of degraded samples. However, if SNPs are to become a realistic Supplement to current short tandem repeat (STR) typing methods, they must be shown to successfully and reliably analyse the challenging samples commonly encountered in casework situations.The European SNPforID consortium, supported by the EU GROWTH programme, has developed a multiplex of 52 SNPs for forensic analysis, with the amplification of all 52 loci in a single reaction followed by two single base extension (SBE) reactions which are detected with capillary electrophoresis.In order to validate this assay, a variety of DNA extracts were chosen to represent problems such as low copy number and degradation that are commonly seen in forensic casework. A total of 40 extracts were used in the study. each of which was sent to two of the five participating laboratories for typing in duplicate or triplicate. Laboratories were instructed to carry out their analyses as if they were dealing with normal casework samples. Results were reported back to the coordinating laboratory and compared with those obtained from traditional STR typing of the same extracts using Powerplex (R) 16 (Promega). These results indicate that, although the ability to successfully type good quality, low copy number extracts is lower, the 52-plex SNP assay performed better than STR typing on degraded samples, and also on samples that were both degraded and of limited quantity, suggesting that SNP analysis can provide advantages over STR analysis in forensically relevant circumstances. However, there were also additional problems arising from contamination and primer quality issues and these are discussed. (c) 2007 Elsevier Ireland Ltd. All rights reserved.