Comparison between automated DNA extraction employing the EZ1 platform and manual methods using real forensic samples
P. Francez, C. A. F. O. Penido, Gilcelene Do Socorro Medeiros De Brito Costa, Rodrigo Matos De Almeida, Eduarda Emanuelle Silva Pena, K. Funabashi, R. Resque
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Abstract
In the last 30 years, advances in Molecular Biology techniques have allowed its application as an essential tool in the criminal investigation, with a particularly relevant application in the determination of authorship of crimes and the identification of missing persons and unknown corpses. However, with the popularization of Forensic Genetics and the growth of violence, there has been a significant increase in the demand for these tests, which has led Forensic Genetics Laboratories to search for more efficient and safe methods of processing this increasing volume of samples. In this sense, automation was one of the adopted solutions, allowing the processing of the samples more accurately and with the minimum of human interference, reducing the risks, and being a more economical alternative when used in large scale. This paper aims to present the results obtained concerning the standardization of the use of an automated DNA extraction platform and to evaluate the advantages of this method in relation to the manual methods in the extraction of forensic samples from the Laboratory of Forensic Genetics of POLITEC-AP. For the realization of the experiments, 500 samples of DNA were extracted using manual methods (Organic and Chelex 100®) and using the automated extraction equipment EZ1 Advanced XL® from Qiagen ©. The automated extraction using the EZ1® following the manufacturer's protocol or after the adaptation of the protocol presented qualitative and quantitative results superior to those obtained using manual methods, both for reference saliva samples and unknown samples of unknown cadavers (teeth and bones) and secretions collected from victims of sexual violence.
采用EZ1平台的自动DNA提取与使用真实法医样本的人工方法的比较
在过去的30年里,分子生物学技术的进步使其成为刑事调查的一个重要工具,在确定犯罪作者以及识别失踪人员和未知尸体方面有着特别重要的应用。然而,随着法医遗传学的普及和暴力的增长,对这些测试的需求显著增加,这促使法医遗传学实验室寻找更有效、更安全的方法来处理不断增加的样本。从这个意义上说,自动化是采用的解决方案之一,可以更准确地处理样本,并将人为干扰降至最低,降低风险,在大规模使用时是一种更经济的替代方案。本文旨在介绍自动化DNA提取平台使用标准化的结果,并评估该方法与POLITEC-AP法医遗传学实验室法医样本提取手动方法相比的优势。为了实现实验,使用手动方法(Organic和Chelex 100®)和Qiagen©的自动提取设备EZ1 Advanced XL®提取了500个DNA样本。在参考唾液样本和未知尸体(牙齿和骨骼)的未知样本以及从性暴力受害者身上采集的分泌物方面,根据制造商的方案或在方案调整后使用EZ1®进行的自动提取呈现出优于使用手动方法获得的定性和定量结果。
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