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从沉积在皮肤上的人类唾液中提取DNA及其在法医鉴定程序中的应用。

DNA extraction from human saliva deposited on skin and its use in forensic identification procedures.

作者信息

Anzai-Kanto Evelyn, Hirata Mário Hiroyuki, Hirata Rosario Dominguez Crespo, Nunes Fabio Daumas, Melani Rodolfo Francisco Haltenhoff, Oliveira Rogério Nogueira

机构信息

Department of Clinical and Toxicological Analysis, School of Pharmaceutical Sciences, University of São Paulo.

出版信息

Braz Oral Res. 2005 Jul-Sep;19(3):216-22. doi: 10.1590/s1806-83242005000300011. Epub 2005 Nov 21.

Abstract

Saliva is usually deposited in bite marks found in many homicides, assault and other criminal cases. In the present study, saliva obtained from volunteers was deposited on skin and recovered for DNA extraction and typing in order to evaluate its usefulness for practical case investigation and discuss the contribution of forensic dentistry to saliva DNA typing. Twenty saliva samples were collected from different donors and used as suspects' samples. Five of these samples were randomly selected and deposited (250 microl) on arm skin. Saliva was collected from skin using the double swab technique. DNA from saliva and skin-deposited saliva samples was extracted by the phenol-chloroform method. DNA samples were amplified by PCR for DNA typing using a set of 15 STRs. The recovery of DNA from saliva deposited in the skin was 14 to 10 times lower than DNA quantity from saliva samples. DNA typing was demonstrated in 4 of 5 deposited saliva samples, the likelihood ratios estimated for these samples based on data of the Brazilian population were 1:11, 1:500, 1:159.140 and 1:153.700.123. Our results indicate that standardized procedures used for DNA collection and extraction from skin-deposited saliva can be used as a method to recover salivary DNA in criminal cases. However, it is important to observe that DNA recovery in forensic samples can be difficult. This study suggests that the analysis of saliva deposited on skin be incorporated into a criminal investigation since it may have great discriminatory power.

摘要

在许多凶杀案、袭击及其他刑事案件中发现的咬痕上通常会留有唾液。在本研究中,从志愿者处获取唾液并将其涂抹在皮肤上,然后进行回收以提取DNA并进行分型,目的是评估其在实际案件调查中的实用性,并探讨法医牙科学对唾液DNA分型的贡献。从不同捐赠者处收集了20份唾液样本,并将其用作嫌疑人样本。其中5份样本被随机选取并(250微升)涂抹在手臂皮肤上。使用双拭子技术从皮肤上收集唾液。采用酚-氯仿法从唾液及皮肤涂抹唾液样本中提取DNA。使用一组15个短串联重复序列(STR)通过聚合酶链反应(PCR)对DNA样本进行扩增以进行DNA分型。从皮肤涂抹唾液中回收的DNA量比唾液样本中的DNA量低14至10倍。在5份涂抹唾液样本中有4份成功进行了DNA分型,根据巴西人群数据估算的这些样本的似然比分别为1:11、1:500、1:159140和1:153700123。我们的结果表明,用于从皮肤涂抹唾液中收集和提取DNA的标准化程序可作为刑事案件中回收唾液DNA的一种方法。然而,必须注意的是,法医样本中的DNA回收可能具有挑战性。本研究表明,对涂抹在皮肤上的唾液进行分析应纳入刑事调查,因为它可能具有很强的鉴别力。

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