Hanaoka Y, Minaguchi K
Department of Forensic Odontology, Tokyo Dental College, Chiba, Japan.
J Forensic Sci. 1996 Sep;41(5):855-8.
Sex determination from blood and teeth by PCR amplification of the alphoid satellite family according to the method of Witt and Erickson was investigated. Amplification of the X-(131 bp) and Y-(172 bp) specific sequences in males and that of the X-specific sequence in females was satisfactory in almost all samples. However, of the 22 samples (blood: 5, dental pulp: 12, tooth hard tissue: 5), a false positive fragment was detected at 172 bp among the female samples (blood: 17, dental pulp: 30, tooth hard tissue: 23) as a result of silver staining. These bands were not amplified by the dual PCR method and were different from the Y-specific band. Subsequent restriction enzyme digestion (DdeI, HinfI, and NciI) of these bands resulted in different patterns from the typical Y-specific band. Furthermore, in five male samples of hard tooth tissue specimens (n = 14), a Y-specific band was detected as a weak band, and in two of these five samples, this weak specific band was not amplified by dual PCR in spite of the male origin of the samples. These results showed that this method is useful for sex determination, but that dual PCR and restriction analysis should be used together in practical applications.
按照维特和埃里克森的方法,通过对α卫星家族进行聚合酶链反应(PCR)扩增,对血液和牙齿样本进行性别鉴定展开了研究。在几乎所有样本中,男性样本中X特异性序列(131bp)和Y特异性序列(172bp)的扩增以及女性样本中X特异性序列的扩增效果均令人满意。然而,在22个样本(血液样本5个、牙髓样本12个、牙齿硬组织样本5个)中,经银染后,在女性样本(血液样本17个、牙髓样本30个、牙齿硬组织样本23个)中检测到一条172bp的假阳性片段。这些条带不能通过双重PCR方法扩增,且与Y特异性条带不同。对这些条带进行后续的限制性酶切消化(DdeI、HinfI和NciI),得到的图谱与典型的Y特异性条带不同。此外,在14个硬齿组织样本中的5个男性样本中,检测到一条Y特异性条带为弱条带,在这5个样本中的2个样本中,尽管样本来源于男性,但这条弱特异性条带不能通过双重PCR扩增。这些结果表明,该方法对性别鉴定有用,但在实际应用中应将双重PCR和限制性分析结合使用。