Identification of Body Fluid Stains Using Real-time RT-PCR: Discrimination Between Salivary, Nasal, and Vaginal Secretions
Identification of Body Fluid Stains Using Real-time RT-PCR: Discrimination Between Salivary, Nasal, and Vaginal Secretions
复制标题
使用实时 RT-PCR 鉴定体液染色：区分唾液、鼻腔和阴道分泌物
DOI：
10.3408/jafst.18.1
复制
发表时间：
2013
期刊：
影响因子：
--
通讯作者：
K. Kasai
中科院分区：
文献类型：
--
作者：
Koichi Sakurada;T. Akutsu;Ken Watanabe;S. Miyasaka;K. Kasai
在刑事调查中，必须澄清从现有生物样本中提取的DNA的来源。近年来，一种新的鉴定体液斑的方法被证明是可行的，它涉及到特异性mRNA表达水平的比较。在这里，我们使用实时RT-PCR检测唾液、鼻腔和阴道分泌物中STATH、HTN 3、MUC 4和ESR 1基因的表达水平，这些基因通常被认为难以区分。STATH在所有唾液（dCt=1.27±3.18，n= 10）和所有鼻分泌物（dCt=-0.99 ~3.94，n=8）中均呈高水平表达，而在10种阴道分泌物中有3种呈低水平表达（dCt=16.34-17.47）。HTN 3在所有唾液分泌物中均以高水平表达（dCt=0.67±3.08）。MUC 4在所有阴道分泌物中均以相对高水平表达（dCt=4.97±2.36，n=10），在10种唾液分泌物中的5种（dCt=8.55-10.40）和8种鼻分泌物中的4种（dCt =6.56-8.23）中以中等水平表达。ESR 1在所有阴道分泌物中均以相对较高的水平表达（dCt=5.79±2.90），而在唾液和鼻腔样品中未发现表达。这些结果表明，不同体液之间的特异性鉴别是可能的。为了测试其实用性，在唾液、精液和血迹中检查了基因敏感性和稳定性。几个基因，包括ACTB，STATH，HTN 3，PRM 2，SEMG 1和HBB，被证明是可检测的，即使在小体积（0.1 mL）的污渍，也表达在1岁的污渍。mRNA的稳定性依赖于有利的环境因素，如干燥和遮光。此外，成功地检测目标基因的混合菌株和模拟的个案样本可能会承诺该检测的实用性。因此，mRNA检测似乎是一种新的工具，从生物样品中的体液鉴定。
In a criminal investigation, it is important to clarify the origin of the extracted DNA from available biological samples. In recent years, a new approach for the identiˆcation of body ‰uid stains has been demonstrated and involves a comparison of speciˆc mRNA expression levels. Here, we used real-time RT-PCR to examine the expression levels of STATH, HTN3, MUC4, and ESR1 genes from salivary, nasal, and vaginal secretions which are often thought to be di‹cult to discriminate between. STATH was expressed at high levels in all salivary (dCt=1.27±3.18, n= 10) and all nasal secretions (dCt=-0.99±3.94, n=8), and at low levels in 3 of the 10 vaginal secretions (dCt=16.34-17.47). HTN3 was expressed at high levels in all salivary secretions (dCt=0.67±3.08). MUC4 was expressed at relatively high levels in all vaginal secretions (dCt=4.97±2.36, n=10), and at moderate levels in 5 of the 10 salivary secretions (dCt=8.55-10.40) and in 4 of the 8 nasal secretions (dCt =6.56-8.23). ESR1 was expressed at relatively high levels in all vaginal secretions (dCt=5.79±2.90), but no expression was found in salivary and nasal samples. These results indicate the possibility for speciˆc discrimination between various body ‰uids. To test the practicality of this, gene sensitivity and stability were examined in saliva, semen, and blood stains. Several genes, including ACTB, STATH, HTN3, PRM2, SEMG1, and HBB, were shown to be detectable even in small volume (0.1 mL) stains and were also expressed in 1-year-old stains. mRNA stability was dependent on favorable environmental factors, such as dryness and shading. In addition, the successful detection of target genes from mixed stains and simulated casework samples might promise the utility of this assay. The mRNA assay therefore appears to be a novel tool for body ‰uid identiˆcation from biological samples.