Direct amplification of casework bloodstains using the Promega PowerPlex® 21 PCR Amplification System
Direct amplification of casework bloodstains using the Promega PowerPlex® 21 PCR Amplification System
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DOI：
10.1016/j.fsigen.2014.05.003
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发表时间：
2014-09-01
影响因子：
3.1
通讯作者：
Scott, Pam
中科院分区：
文献类型：
--
作者：
Gray, Kerryn;Crowle, Damian;Scott, Pam
提交给塔斯马尼亚法医科学服务局（FSST）的大量证据是血液拭子或血渍物品。这些物品的样品在Powerplant（R）21扩增之前常规地进行苯酚：氯仿：异戊醇有机提取和定量聚合酶链反应（qPCR）检测。在政府预算紧缩的财政环境下，这种多步骤的过程具有显著的成本和时间表影响，提高运营效率的压力，以及越来越强调更好地支持情报主导的警务的快速技术。使用Powerplant（R）21在布和Whatman FTA（TM）卡上直接扩增血液和口腔细胞已经成功地用于参考样本，消除了对样品预处理的要求。扩大这种方法的范围，以包括不太原始的个案血液拭子和血渍物品的样品进行了探索，努力消除冗长的DNA提取，纯化和qPCR步骤，为更广泛的样本子集。血液沉积在一系列基板上，包括那些历史上发现抑制STR扩增。使用微穿孔、微拭子或两者采集样本。通过在全体积和减少体积（25和13 μ L）下扩增所有样品，评估了通过减少体积扩增进一步节省财政的潜力。总体成功率数据显示，80%的样品在减少体积下产生了完整的轮廓，而在全体积下为78%。织物/纺织品上血液的成功率特别高，100%的微穿孔样本在减少的体积下产生完整的轮廓，在全体积下产生85%的轮廓。已经经历了显着的好处，最明显的是在采访嫌疑人之前向警方调查人员提供了关键物品的结果，并且在很短的时间内成功完成了验尸，以避免延迟将遗骸释放给家人。(C)2014爱思唯尔爱尔兰有限公司版权所有。
A significant number of evidence items submitted to Forensic Science Service Tasmania (FSST) are blood swabs or bloodstained items. Samples from these items routinely undergo phenol: chloroform: isoamyl alcohol organic extraction and quantitative Polymerase Chain Reaction (qPCR) testing prior to PowerPlex (R) 21 amplification. This multi-step process has significant cost and timeframe implications in a fiscal climate of tightening government budgets, pressure towards improved operating efficiencies, and an increasing emphasis on rapid techniques better supporting intelligence-led policing.Direct amplification of blood and buccal cells on cloth and Whatman FTA (TM) card with PowerPlex (R) 21 has already been successfully implemented for reference samples, eliminating the requirement for sample pre-treatment. Scope for expanding this method to include less pristine casework blood swabs and samples from bloodstained items was explored in an endeavour to eliminate lengthy DNA extraction, purification and qPCR steps for a wider subset of samples.Blood was deposited onto a range of substrates including those historically found to inhibit STR amplification. Samples were collected with micro-punch, micro-swab, or both. The potential for further fiscal savings via reduced volume amplifications was assessed by amplifying all samples at full and reduced volume (25 and 13 mu L).Overall success rate data showed 80% of samples yielded a complete profile at reduced volume, compared to 78% at full volume. Particularly high success rates were observed for the blood on fabric/textile category with 100% of micro-punch samples yielding complete profiles at reduced volume and 85% at full volume.Following the success of this trial, direct amplification of suitable casework blood samples has been implemented at reduced volume. Significant benefits have been experienced, most noticeably where results from crucial items have been provided to police investigators prior to interview of suspects, and a coronial identification has been successfully completed in a short timeframe to avoid delay in the release of human remains to family members. (C) 2014 Elsevier Ireland Ltd. All rights reserved.