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使用基于mRNA的检测方法鉴定犬类唾液。

Identification of canine saliva using mRNA-based assay.

作者信息

Nakanishi Hiroaki, Ohmori Takeshi, Hara Masaaki, Yoneyama Katsumi, Takada Aya, Saito Kazuyuki

机构信息

Department of Forensic Medicine, Juntendo University School of Medicine, 2-1-1, Hongo, Bunkyo-Ku, Tokyo, 113-8421, Japan.

National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa, Chiba, 277-0882, Japan.

出版信息

Int J Legal Med. 2017 Jan;131(1):39-43. doi: 10.1007/s00414-016-1391-7. Epub 2016 May 25.

Abstract

A dog saliva analysis in addition to a bite-mark analysis may be important for evidence when a crime involves a dog bite. In this study, the utility of detecting canine saliva-specific mRNAs to identify canine saliva was evaluated. Canine saliva swabs (n = 20), urine swabs (n = 20), body surface swabs (n = 20), whole blood samples (n = 10), human saliva (n = 20), human skin surface swabs (n = 20), and human whole blood (n = 20) were tested. The saliva-specific genes encoding statherin (STATH), carbonic anhydrase VI (CA-VI), and dog allergens (Canf1 and Canf2) were analyzed as candidate genes. Moreover, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as confirmation of canine mRNA extraction. STATH, CA-VI, Canf1, Canf2, and GAPDH mRNAs were detected in 19/20, 1/20, 11/20, 4/20, and 20/20 saliva samples, respectively. The STATH, CA-VI, Canf1, Canf2, and GAPDH mRNAs did not exhibit cross-reactivity with samples of human origin. This mRNA-based assay was also able to detect canine saliva in mock forensic samples. The results of this study indicated that the detection of STATH mRNA is useful for the identification of canine saliva, and GAPDH is a suitable marker for canine mRNA extraction.

摘要

当犯罪涉及狗咬事件时,除咬痕分析外,狗唾液分析可能对证据收集很重要。在本研究中,评估了检测犬唾液特异性mRNA以鉴定犬唾液的效用。对犬唾液拭子(n = 20)、尿液拭子(n = 20)、体表拭子(n = 20)、全血样本(n = 10)、人类唾液(n = 20)、人类皮肤表面拭子(n = 20)和人类全血(n = 20)进行了检测。分析了编码富组蛋白(STATH)、碳酸酐酶VI(CA-VI)和犬过敏原(Canf1和Canf2)的唾液特异性基因作为候选基因。此外,使用甘油醛-3-磷酸脱氢酶(GAPDH)来确认犬mRNA的提取。STATH、CA-VI、Canf1、Canf2和GAPDH mRNA在20个唾液样本中的检出率分别为19/20、1/20、11/20、4/20和20/20。STATH、CA-VI、Canf1、Canf2和GAPDH mRNA与人类来源的样本未表现出交叉反应。这种基于mRNA的检测方法也能够在模拟法医样本中检测到犬唾液。本研究结果表明,检测STATH mRNA对鉴定犬唾液有用,GAPDH是犬mRNA提取的合适标志物。

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