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利用软组织中的 DNA 进行高度腐烂尸体的遗传鉴定。

Genetic identification of highly putrefied bodies using DNA from soft tissues.

机构信息

Department of Legal Medicine, University Hospital of Schleswig-Holstein, Arnold-Heller-Straße 12, Kiel, Germany.

出版信息

Int J Legal Med. 2011 Nov;125(6):891-4. doi: 10.1007/s00414-010-0537-2. Epub 2010 Dec 18.

Abstract

The identification of putrefied bodies is a common task in forensic routine work. The deceased are usually identified by dental records, fingerprinting, or--in cases where no such data are available--DNA analysis. However, with progressive putrefaction, DNA integrity is rapidly decreasing. Genetic analysis may then be greatly impaired, if not impossible. The aim of our study was to establish an efficient procedure to successfully extract and amplify DNA from soft tissues of bodies in different stages of putrefaction. Soft tissues-unlike teeth or bones-usually allow the application of fast and easy-to-use extraction protocols. DNA was extracted from different tissues (aorta, kidney, liver, and skeletal muscle) taken at autopsy using a commercially available DNA extraction kit, and DNA quality and quantity were controlled by agarose gel electrophoresis and real-time polymerase chain reaction (PCR). Presence of mitochondrial DNA was tested using a highly sensitive duplex PCR. Short tandem repeat analysis was done using the AmpFlSTR Identifiler kit. Additionally, mitochondrial DNA sequencing was performed. After DNA extraction from at least two different tissues-preferably the kidney and the aorta-with the extraction kit based on the Nucleobond method, a successful amplification of at least eight loci was possible in 17 out of 18 cases, and 12 or more loci could be amplified in 15 cases.

摘要

尸体身份识别是法医学常规工作中的一项常见任务。通常通过牙齿记录、指纹识别,或者——在没有这些数据的情况下——通过 DNA 分析来识别死者。然而,随着腐烂的进展,DNA 的完整性迅速下降。如果不是不可能的话,那么遗传分析可能会受到极大的损害。我们的研究目的是建立一种有效的程序,以成功地从不同腐烂阶段的尸体软组织中提取和扩增 DNA。与牙齿或骨骼不同,软组织通常允许应用快速且易于使用的提取方案。使用市售的 DNA 提取试剂盒从尸检时获得的不同组织(主动脉、肾脏、肝脏和骨骼肌)中提取 DNA,并通过琼脂糖凝胶电泳和实时聚合酶链反应 (PCR) 控制 DNA 质量和数量。使用高度敏感的双重 PCR 测试线粒体 DNA 的存在。使用 AmpFlSTR Identifiler 试剂盒进行短串联重复分析。此外,还进行了线粒体 DNA 测序。在用基于 Nucleobond 方法的提取试剂盒从至少两个不同的组织(最好是肾脏和主动脉)中提取 DNA 后,在 18 个案例中的 17 个案例中可以成功扩增至少 8 个基因座,并且在 15 个案例中可以扩增 12 个或更多基因座。

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