Development of a Real-Time PCR-Based Method for Analyzing Semen-Specific Unmethylated DNA Regions and Methylation Status in Aged Body Fluid Stains
Development of a Real-Time PCR-Based Method for Analyzing Semen-Specific Unmethylated DNA Regions and Methylation Status in Aged Body Fluid Stains
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开发基于实时 PCR 的方法来分析老化体液染色中精液特异性非甲基化 DNA 区域和甲基化状态
DOI：
10.1111/1556-4029.12941
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发表时间：
2016
影响因子：
1.6
通讯作者：
Koichi Sakurada
中科院分区：
文献类型：
--
作者：
Ken Watanabe;Tomoko Akutsu;Koichi Sakurada
在法医调查中，精液的检测被认为是性侵犯的重要证据。在这项研究中，我们报告了一种基于真实的实时聚合酶链反应的方法，用于鉴定精液，可以简单快速地分析精液特异性的DACT 1基因的非甲基化区域。该方法使用两种针对甲基化或非甲基化状态设计的荧光探针，可以对该区域的甲基化状态进行定量分析。此外，该方法还用于分析各种体液样本，包括29岁的精液和血迹。结果表明，该方法能检测到几乎全部的精液或非精液信号，即使在高度分解的样品中，也能检测到其它荧光探针的微弱信号。虽然仍然需要进一步的分析，例如设置阈值来分析未知样本，但这种方法可能是识别精液的有用补充工具，特别是在旧污渍中，例如在冷案调查中。
The detection of semen in forensic investigation is considered important evidence of sexual assault. In this study, we report the development of a real‐time polymerase chain reaction‐based method for identifying semen that can simply and rapidly analyze the semen‐specific unmethylated region of theDACT1gene. Using two fluorescent probes designed for the methylated or unmethylated status, this method could perform quantitative analysis of the methylation status in this region. Furthermore, this method was used to analyze various body fluid samples, including 29‐year‐old semen and blood stains. The results showed that this method can detect almost exclusively semen or nonsemen signals even in highly decomposed samples, while a few semen or nonsemen samples showed slight signals of the other fluorescence probe. Although there is still a need for further analysis such as setting thresholds to analyze unknown samples, this method could be a useful supplementary tool for identifying semen, especially in old stains such as those in cold‐case investigations.