Extraction of DNA from decomposed human tissue -: An evaluation of five extraction methods for short tandem repeat typing
Extraction of DNA from decomposed human tissue -: An evaluation of five extraction methods for short tandem repeat typing
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DOI：
10.1016/s0379-0738(99)00128-0
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发表时间：
1999-11-08
影响因子：
2.2
通讯作者：
Olaisen, B
中科院分区：
文献类型：
--
作者：
Hoff-Olsen, P;Mevåg, B;Olaisen, B
超多态性短串联重复DNA序列，STR或微卫星，已被广泛用于人类身份识别，特别是在刑事案件和大规模灾难中。在这种情况下，分析的基质可能是分解的生物材料，这是在选择适当的个案工作方法时必须考虑的一个事实。在本文中，我们报告了五种不同的DNA提取方法，即苯酚-氯仿，二氧化硅为基础的，InstaGene矩阵（TM）（BioTest），玻璃纤维过滤器，和Chelex为基础的方法的评价。分析的基质是法医尸检病例中分解的人肝组织标本。提取的DNA进行定量和DNA分析的一组七个STR。我们比较了实验室的时间消耗和成本的五种方法，表明Chelex方法是更快速，更便宜的方法，苯酚-叶绿素和二氧化硅提取是最耗时和资源需求的。在10例中有9例通过二氧化硅方法获得了完整的轮廓，在70个STR分析中有4个未能给出可靠的类型。在70个STR分析中，酚-叶绿素法和玻璃纤维过滤法在16个分析中失败，InstaGene Matrix（TM）（BioTest）在25个分析中失败，Chelex提取物在56个分析中失败。通过多元Logistic回归分析，我们发现二氧化硅程序与其他方法之间的差异具有统计学意义。在我们手中，当可用的生物材料是分解的人体组织时，硅胶提取程序是一个明显的选择，即使这个程序是比较费力的程序之一。(C)1999爱思唯尔科学爱尔兰有限公司保留所有权利。
Hyperpolymorphic short tandem repetitive DNA sequences, STRs or microsatellites, have become widely used in human identification, particularly in criminal cases and in mass disasters. In such cases the substrates for the analyses may he decomposed biological material, a fact that has to be taken into account when choosing the appropriate casework methods. In this paper we report the evaluation of five different DNA extraction methods, namely the phenol-chloroform, the silica based, the InstaGene Matrix(TM) (BioTest), the glass fiber filter, and the Chelex based methods. The substrates for the analyses are decomposed human liver tissue specimens from forensic autopsy cases. Extracted DNA was quantified and DNA profiled by a set of seven STRs. We have compared laboratory time consumption and costs of the five methods, showing that the Chelex method is the more rapid and less expensive of the methods, the phenol-chlorophorm and silica extractions being the most time consuming and resource demanding ones. A full profile was obtained by the silica method in nine out of ten cases and this method failed to give a reliable type in four out of 70 STR analyses. The phenol-chlorophorm and the glass fiber filter methods failed in 16 analyses, the InstaGene Matrix(TM) (BioTest) in 25 and the Chelex extracts in 56 of the 70 STR analyses. By multiple logistic regression we show that the difference between the silica procedure and the other methods are statistically significant. In our hands, the silica gel extraction procedure is an obvious choice when the biological material available is decomposed human tissue even if this procedure is one of the more laborious ones. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.