重组酶聚合酶扩增技术在现场物证唾液鉴定中的应用  

Application of Recombinase Polymerase Amplification Technology in the Identification of Biological Evidence in Salivary Spots

作　　者：许炎[1,2] 曹禹 郝思静[1] 黄江平 王旭芳[3] 吴丹 XU Yan;CAO Yu;HAO Sijing;HUANG Jiangping;WANG Xufang;WU Dan(Key Laboratory of Forensic Evidence and Science Technology,Ministry of Public Security,Institute of Forensic Science,Shanghai Public Security Bureau,Shanghai 200083,China;Shanghai Police College,Shanghai 201210,China;Zhejiang Zhonghe Institute of Forensic Science,Ningbo 315300,China)

机构地区：[1]上海市公安局物证鉴定中心,法医物证学现场应用技术公安部重点实验室,上海200083 [2]上海公安学院,上海201210 [3]浙江中和司法鉴定中心,浙江宁波315300

出　　处：《中国司法鉴定》2022年第6期57-60,共4页Chinese Journal of Forensic Sciences

基　　金：公安部应用创新计划项目(2018YYCXSHSJ027)。

摘　　要：目的建立一种基于重组酶聚合酶扩增(RPA)技术的现场唾液生物属性鉴定方法。方法利用DNA-RNA共提取技术,将样本总RNA反转录成c DNA,针对唾液特异性分子标识HTN3设计RPA引物,构建HTN3 RPA检测方法,并测试该方法的特异性和应用效果。结果反转录产物在39℃恒温反应20min即可完成目的片段的有效扩增。扩增条带与目的序列完全一致。在对20份现场检材的检验中该方法均获得了稳定可靠的检测结果。结论建立的反转录RPA检测唾液生物属性方法特异性强,极大缩短了扩增时间,结果稳定且易于判定,适合实验室快速检测。该方法可同步完成物证的STR分型检验,为丰富法医物证体液鉴定提供了一种新的策略和方法。Objective To establish an on-site biological body-fluids identification method in saliva samples based on Recombinase Polymerase Amplification(RPA)technology.Methods Using DNA-RNA co-extraction technology,the total RNA of the sample was reverse-transcribed into cDNA.RPA primers were designed for the saliva-specific gene marker HTN3 to construct a HTN3 RPA detection method,and the specificity and application effect of the method were tested.Results The reverse transcription product can be reacted at 39℃for 20 min to complete the effective amplification of the target fragment.The amplified band was completely consistent with the target sequence.This method has obtained stable and reliable results by testing of 20 on-site biological body-fluids materials.Conclusion This study established a reverse transcription RPA method for the detection of salivary spots.It has strong specificity,and will greatly shorten the amplification time.The results were stable and easy to be determined,which is suitable for rapid laboratory testing.The STR typing of biological evidence can be completed simultaneously.The method provides a new strategy for forensic body fluid identification.

关 键 词：法医物证学 重组酶聚合酶扩增 唾液 体液鉴定 

分 类 号：DF795.4[医药卫生—法医学]