Sakurada Koichi, Ikegaya Hiroshi, Fukushima Hisayo, Akutsu Tomoko, Watanabe Ken, Yoshino Mineo
National Research Institute of Police Science, 6-3-1, Kashiwanoha, Kashiwa, Chiba 277-0882, Japan.
Leg Med (Tokyo). 2009 May;11(3):125-8. doi: 10.1016/j.legalmed.2008.10.002. Epub 2008 Nov 25.
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 microL 6-day-old stains, and showed high specificity in 7-day-old 30 microL 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.
近年来，逆转录聚合酶链反应（RT-PCR）多重mRNA分析已作为一种鉴定体液的新方法被报道。我们也已经证明了使用实时RT-PCR检测鉴定体液的可行性。选定用于检测唾液的基因——磷蛋白（STATH）和富组蛋白（HTN3），以及选定用于检测精液的基因——鱼精蛋白2（PRM2）和精液蛋白1（SEMG1），对这些体液显示出高度特异性。因此，我们在体液污渍中检测了目标基因的敏感性和特异性。在0.1微升6日龄的污渍中检测到了所有目标基因，并且在7日龄的30微升污渍中显示出高特异性。此外，我们还研究了HTN3在唾液污渍中在不同环境条件下随时间的稳定性。结果表明，污渍中mRNA的降解受潮湿条件的影响很大，并且光照也是一个重要因素。然而，在保存于干燥和黑暗条件下的陈旧唾液污渍（6年）和陈旧精液污渍（3.5年）中仍可检测到mRNA。超出我们预期的mRNA稳定性可能在开发体液鉴定新技术中发挥重要作用。
