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使用热启动聚合酶链反应-序列特异性引物对无名尸体的主动脉组织进行HLA分型。

HLA typing of aortic tissues from unidentified bodies using hot start polymerase chain reaction-sequence specific primers.

作者信息

Sato Yayoi, Hayakawa Mutsumi, Nakajima Toru, Motani Hisako, Kiuchi Masahiro

机构信息

Department of Legal Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba City, Chiba 260-8670, Japan.

出版信息

Leg Med (Tokyo). 2003 Mar;5 Suppl 1:S191-3. doi: 10.1016/s1344-6223(02)00108-6.

Abstract

This study investigated the identification of unidentified bodies through HLA DNA typing of aortic tissues. Eight aortas were collected at autopsy from six bodies found in water, one burnt body, and one mummified body, respectively. DNA was extracted from 10-20 mg of aortic tissue with a PUREGENE DNA Isolation Kit and phenol/chloroform. HLA (A, B, DR or DQ locus) alleles were typed using the hot start polymerase chain reaction-sequence specific primers (PCR-SSP) method with a Dynal AllSet(+) SSP "low resolution" kit for each locus. The aorta was still retained in degraded samples, those from the drowned, the burnt, and the mummified bodies. In each case, approximately 0.04-3.84 microg of DNA was recovered from 1 mg of fixed tissue. We typed 8/8 for the DR, 4/4 for the DQ, 3/3 for the A, and 3/3 for the B locus from the samples. Based on these results and the finding that DNA extraction is easier from the aorta than from other samples such as bone or tooth, we considered this method to be useful for forensic samples.

摘要

本研究通过对主动脉组织进行HLA DNA分型来调查无名尸体的身份鉴定。分别从水中发现的6具尸体、1具烧焦尸体和1具木乃伊尸体的尸检中收集了8个主动脉。使用PUREGENE DNA分离试剂盒和苯酚/氯仿从10 - 20mg主动脉组织中提取DNA。使用热启动聚合酶链反应 - 序列特异性引物(PCR - SSP)方法,采用Dynal AllSet(+) SSP“低分辨率”试剂盒对每个位点的HLA(A、B、DR或DQ位点)等位基因进行分型。在降解样本中,即来自溺水者、烧焦尸体和木乃伊尸体的样本中,主动脉仍然保留。在每种情况下,从1mg固定组织中回收了约0.04 - 3.84μg DNA。我们对样本的DR位点分型成功率为8/8,DQ位点为4/4,A位点为3/3,B位点为3/3。基于这些结果以及从主动脉中提取DNA比从骨骼或牙齿等其他样本中更容易的发现,我们认为这种方法对法医样本有用。

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