Evaluation of mRNA-based approach for identification of saliva and semen
Evaluation of mRNA-based approach for identification of saliva and semen
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DOI：
10.1016/j.legalmed.2008.10.002
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发表时间：
2009-05-01
期刊：
影响因子：
1.5
通讯作者：
Yoshino, Mineo
中科院分区：
文献类型：
--
作者：
Sakurada, Koichi;Ikegaya, Hiroshi;Yoshino, Mineo
在过去的几年中，通过逆转录-聚合酶链反应（RT-PCR）进行的多重mRNA分析已被报道为用于鉴定体液的新方法。我们还证明了通过使用实时RT-PCR测定来识别体液的可行性。唾液中选择的Statherin（STATH）和histatin（HTN 3）以及精液中选择的鱼精蛋白2（PRM 2）和精液凝固蛋白1（SEMG 1）对这些体液表现出高度特异性。因此，在体液污渍中检查靶基因的敏感性和特异性。在0.1 μ L的6日龄菌株中检测到所有目的基因，在7日龄30 μ L菌株中显示出高特异性。此外，在各种环境条件下随时间检查了HTN 3在唾液斑中的稳定性。结果表明，湿环境对菌株中mRNA的降解有很大影响，光照也是一个重要因素。然而，mRNA在较老的唾液染色（6岁）和较老的精液染色（3.5岁）中可检测到，这两种染色都保持在干燥和黑暗的条件下。mRNA的稳定性超出了我们的假设，可能在开发体液鉴定新技术中发挥重要作用。(C)2008爱思唯尔爱尔兰有限公司保留所有权利。
Multiplex mRNA profiling by a reverse transcription-polymerase chain reaction (RT-PCR) has been reported in the last few years as a new approach for the identification of body fluids. We have also demonstrated the feasibility of identifying body fluids by using a real-time RT-PCR assay. Statherin (STATH) and histatin (HTN3), the selected genes for saliva, and protamin 2 (PRM2) and semenogelin 1 (SEMG1), those selected for semen, showed high specificity to these body fluids. Thus, the sensitivity and specificity of target genes were examined in body fluid stains. All target genes were detected in 0.1 mu L 6-day-old stains, and showed high specificity in 7-day-old 30 mu L stains. Furthermore, the stability of HTN3 in saliva stains was examined under various environmental conditions over time. The results showed that the degradation of mRNA in the stains was highly affected by wet conditions, and that light was also an important factor. However, mRNA was detectable in an older saliva stain (6 years old) and in an older semen stain (3.5 years old), both of which had been kept under dry and dark conditions. The stability of mRNA beyond our supposition may play an important role in developing new techniques for body fluid identification. (C) 2008 Elsevier Ireland Ltd. All rights reserved.