Evaluation of Various Methods of DNA Analysis for Hair Samples

H. Matsuda, K. Sekiguchi, K. Kasai, M. Yoshino, S. Seta
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引用次数: 11

Abstract

Three Kinds of DNA analyses; AmpliType® PM typing, TH01 typing and mitochondrial DNA (mtDNA) analysis were evaluated to establish the corroborative method for forensic hair comparison. Five scalp telogen hairs were collected from 12 Japanese males ranging in age from 26 to 33 years. DNA was extracted from five hair shafts of 5cm in length and from five hair roots of 3mm in length taken from each subject. The PM typing was performed using the AmpliType® PM PCR Amplification and Typing Kit. The TH01 typing was carried out using Quick-TypeTM HUMTH01 and was detected by silver staining. For the mtDNA analysis, the sequencings of the two hypervariable regions (HV1 and HV2) of control region were performed by using the ABI PRISMTM Dye Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq® DNA polymerase FS and ABI PRISMTM model 377 DNA sequencer.   In the telogen hair, the hair root was a suitable sample for both PM and TH01 typing compared to the hair shaft samples (about 50% of the subjects could be detected from the hair root samples). The PM typing and TH01 typing showed almost equal detectability. In the mtDNA analysis, the PCR amplifications of HV1 and HV2 were successfully performed in all twelve subjects by employing new primer, and the sequences of the PCR products from two subjects were determined. This result suggests that the mtDNA analysis can be applied for hair comparison in cases where the genomic DNA typing is not available. However, further study for the reliability and reproducibility of the mtDNA analysis should be performed.
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各种毛发样本DNA分析方法的评价
三种DNA分析方法;对AmpliType®PM分型、TH01分型和线粒体DNA (mtDNA)分析进行评估,以建立法医头发比较的确证方法。从12名年龄在26岁至33岁之间的日本男性中收集了5根头皮上的休止期头发。从每个受试者的5根长5cm的毛干和5根长3mm的发根中提取DNA。使用AmpliType®PM PCR扩增分型试剂盒进行PM分型。采用Quick-TypeTM HUMTH01进行TH01分型,银染色检测。mtDNA分析,对照区两个高变区(HV1和HV2)的测序采用ABI PRISMTM染料终止周期测序准备反应试剂盒(含AmpliTaq®DNA聚合酶FS)和ABI PRISMTM 377型DNA测序仪。在休止期头发中,与毛干样本相比,发根是PM和TH01分型的合适样本(大约50%的受试者可以从发根样本中检测到)。PM分型与TH01分型检出率基本相等。在mtDNA分析中,采用新的引物成功扩增了12个被试的HV1和HV2病毒,并确定了两个被试的PCR产物的序列。这一结果表明,在基因组DNA分型不可用的情况下,mtDNA分析可用于头发比较。然而,mtDNA分析的可靠性和可重复性有待进一步研究。
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