Practical evaluation of an RNA-based saliva identification method
Practical evaluation of an RNA-based saliva identification method
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DOI：
10.1016/j.scijus.2017.07.001
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发表时间：
2017-11-01
影响因子：
1.9
通讯作者：
Sakurada, Koichi
中科院分区：
文献类型：
--
作者：
Watanabe, Ken;Akutsu, Tomoko;Sakurada, Koichi
鉴定在犯罪现场发现的样本中的唾液对于澄清用于鉴定个人身份的DNA的组织来源非常重要。最近，一种新的基于信使RNA的方法，使用两个唾液特异性标记物，Statherin（STATH）和Histatin 3（HTN 3），已被报道。这种方法可以比传统的基于淀粉酶的方法更特异地识别唾液。在这里，我们进行了几项评估，将这种方法应用于现实世界的法医工作。首先，我们评估了暴露于蓝光（450 nm）或Phadebas纸上的试剂（这是用于定位唾液污渍的直接方法）对RNA标记物稳定性的影响。结果表明，暴露于两种直接检测不影响RNA标记物的稳定性。其次，我们对基于RNA和基于淀粉酶的常规方法进行了比较分析，以检查标记在各种储存条件下的灵敏度和稳定性。虽然这两种方法检测1天大唾液斑的灵敏度没有差异，但一项时间进程研究表明，RNA唾液标记物的稳定性低于淀粉酶，尤其是在潮湿条件下。在该时间过程实验中，还研究了人DNA的稳定性。虽然DNA在潮湿条件下也不稳定，但它在干燥条件下比RNA标记更稳定。考虑到上述结果，我们建议将RNA方法引入到目前的唾液鉴定程序中，并应作为一种补充方法，以有力地支持淀粉酶为基础的方法鉴定唾液。(C)2017年，特许法医学学会。由Elsevier爱尔兰有限公司出版。保留所有权利。
Identifying saliva in samples found at crime scenes is important to clarify the tissue origin of DNA obtained for identification of individuals. Recently, a novel messenger RNA-based approach using two saliva-specific markers, Statherin (STATH) and Histatin 3 (HTN3), has been reported. This method can identify saliva more specifically than conventional amylase-based methods. Here, we performed several evaluations related to applying this method to real-world forensic work. First, we evaluated the effects of exposure to blue light (450 nm) or to the reagent on Phadebas paper, which are direct methods used to locate saliva stains, on the stability of the RNA markers. The results demonstrate that exposure to the two direct tests did not affect the stability of the RNA markers. Second, we performed a comparative analysis of RNA-based and amylase-based conventional methods to examine the sensitivity and stability of the markers under various storage conditions. Although there was no difference in the sensitivity of the two methods for detecting 1-day-old saliva stains, a time-course study demonstrated that the RNA saliva markers were less stable than amylase, especially in wet conditions. During this time-course experiment, the stability of human DNA was also investigated. Although DNA was also unstable in wet conditions, it was more stable than the RNA markers in dry conditions. Taking the above results into consideration, we suggest that the RNA method could be introduced to current saliva identification procedures and should be used as a supplementary method to strongly support identification of saliva by the amylase-based method. (C) 2017 The Chartered Society of Forensic Sciences. Published by Elsevier Ireland Ltd. All rights reserved.