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从指甲中分离外源痕量 DNA 的对比研究。

A comparative study for the isolation of exogenous trace DNA from fingernails.

机构信息

State of California, Department of Justice, Bureau of Forensic Services, 1001 West Cutting Boulevard, Richmond, CA 94804, United States.

State of California, Department of Justice, Bureau of Forensic Services, 1001 West Cutting Boulevard, Richmond, CA 94804, United States.

出版信息

Forensic Sci Int Genet. 2019 Mar;39:119-128. doi: 10.1016/j.fsigen.2018.12.009. Epub 2019 Jan 4.

Abstract

Often fingernails from a victim or suspect involved in a physical assault, such as murder or sexual assault, are submitted to crime laboratories for DNA testing of foreign/exogenous biological material; however, very few studies have been conducted comparing the effectiveness of different sampling methods on the removal of foreign/exogenous DNA while minimizing the fingernail endogenous DNA. In this study three different sampling methods (swabbing, PBS soak, and PrepFiler® lysis buffer soak) were compared in order to identify one that minimizes the amount of endogenous DNA removed and maximizes the amount of foreign/exogenous male DNA removed. The samples were processed using the Tecan HIDEVO150 robot in order to reduce analyst time and the DNA mixtures were interpreted using the probabilistic genotyping software STRmix™. For each sampling method the quantity of male DNA, the mixture proportions, the number of foreign/exogenous male alleles detected, the amount of DNA degradation, and the discrimination power via the likelihood ratio obtained for the foreign/exogenous male DNA donor were determined and compared. The PrepFiler® lysis buffer soak and swabbing sampling methods appear to be equally effective at removing foreign/exogenous DNA from fingernails; however, the lysis buffer soak sampling method extracts more female endogenous DNA from the fingernail and the female DNA is degraded. Marginally higher likelihood ratios were obtained for the swab samples versus the PrepFiler® lysis buffer soak samples; therefore, it was determined that the swabbing sampling method was the best sampling method for the recovery of foreign exogenous DNA from fingernails while minimizing the amount of endogenous DNA removed.

摘要

通常情况下,参与身体攻击(如谋杀或性侵犯)的受害者或嫌疑人的指甲会被提交给犯罪实验室,用于检测外来/外源性生物材料的 DNA;然而,很少有研究比较不同采样方法在去除外来/外源性 DNA 的同时最小化指甲内源性 DNA 的效果。在这项研究中,比较了三种不同的采样方法(拭子、PBS 浸泡和 PrepFiler®裂解缓冲液浸泡),以确定一种可以最小化去除内源性 DNA 量并最大程度去除外来/外源性男性 DNA 的方法。使用 Tecan HIDEVO150 机器人处理样品,以减少分析员的时间,使用概率基因分型软件 STRmix™ 解释 DNA 混合物。对于每种采样方法,确定并比较了男性 DNA 的数量、混合物比例、检测到的外来/外源性男性等位基因数量、DNA 降解程度以及通过获得的外来/外源性男性 DNA 供体的似然比获得的鉴别力。从指甲中去除外来/外源性 DNA 方面,PrepFiler®裂解缓冲液浸泡和拭子采样方法似乎同样有效;然而,裂解缓冲液浸泡采样方法从指甲中提取出更多的女性内源性 DNA,并且女性 DNA 被降解。与 PrepFiler®裂解缓冲液浸泡采样方法相比,拭子采样方法获得的似然比略高;因此,确定拭子采样方法是从指甲中回收外来外源性 DNA 的最佳采样方法,同时最大限度地减少去除的内源性 DNA 量。

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