主管：中华人民共和国司法部
主办：司法鉴定科学研究院
ISSN 1671-2072  CN 31-1863/N
中国司法鉴定

中国司法鉴定 ›› 2025 ›› Issue (6): 38-47.DOI: 10.3969/j.issn.1671-2072.2025.06.005

• 专题研究：法医物证体液/组织溯源 • 上一篇    下一篇

一步法逆转录PCR体液斑鉴定试剂盒的研究与应用测试

王    哲1，段佳瑶2，莫晓婷1，赵一霞1，胡    胜1，董颖强1，詹    飞3，杨    亮4，包赫来5，季安全1，孙启凡1   

  1. 1. 公安部鉴定中心 现场物证溯源技术国家工程实验室 法医遗传学公安部重点实验室，北京 100038；2. 山西医科大学 法医学院，山西 太原 030024；3. 陕西省公安厅，陕西 西安 710000；4. 牡丹江市公安局，黑龙江 牡丹江 157000； 5. 长春市公安局，吉林 长春 130000
  • 收稿日期:2025-05-26 出版日期:2025-11-15 发布日期:2025-11-25
  • 通讯作者: 季安全（1965—），男，主任法医师，硕士，主要从事法医遗传学研究。E-mail：aqjdna@tom.com 孙启凡（1985—），女，副主任法医师，博士，主要从事法医遗传学研究。E-mail：sunqifan@cifs.gov.cn
  • 作者简介:王哲（1997—），男，主要从事法医遗传学研究。E-mail：wangzhe1997@outlook.com
  • 基金资助:
    国家重点研发计划（2022YFC3341002）；公安部科技强警基础工作专项（2022JC11）；中央级公益性科研院所基本科研业务费专项资金项目（2022JB021）；法医遗传学公安部重点实验室开放课题（2022FGKFKT04）。

Development and Testing of a Body Fluid Identification Kit Based on One-Step RT-PCR

WANG Zhe1, DUAN Jiayao2, MO Xiaoting1, ZHAO Yixia1, HU Sheng1, DONG Yingqiang1, #br# ZHAN Fei3, YANG Liang4, BAO Helai5, JI Anquan1, SUN Qifan1#br#   

  1. 1. MPS Key Laboratory of Forensic Genetics, National Engineering Laboratory for Forensic Science, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China; 2. School of Forensic Medicine, Shanxi Medical University, Taiyuan 030024, China; 3. Shaanxi Provincial Public Security Bureau, Xi’an 710000, China; 4. Mudanjiang Public Security Bureau, Mudanjiang 157000, China; 5. Changchun Public Security Bureau, Changchun 130000, China
  • Received:2025-05-26 Published:2025-11-15 Online:2025-11-25

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摘要： 目的 开发一种可快速鉴定常见法医体液斑体液来源的试剂盒。方法 通过筛选特异性mRNA、设计多重引物并优化反应体系等，构建基于一步法逆转录聚合酶链反应（reverse transcription polymerase chain reaction，RT-PCR）技术，可同时进行12种RNA检测的体系。使用收集的5种法医常见体液样本275份及案件检材10份，对试剂盒进行灵敏度、特异度、重复性等测试。通过合成质粒制备阳性标准品，完善试剂盒组分。结果 试剂盒能够成功检测12种RNA，检测时长约为3 h，部分类型样本灵敏度可达到0.02 ng，重复性良好，能够从混合样本中成功检出各组分特异性mRNA，对于案件样本同样具有良好的适用性。结论 本试剂盒可用于鉴别外周血、月经血、精液、唾液和阴道分泌物5种常见体液斑，具有操作简单、耗时短、混合样本鉴别能力强等优势，从而具有良好的法医实战应用前景。

关键词: 法医物证学, mRNA, 体液鉴定, 一步法RT-PCR

Abstract: Objective To develop a kit for rapid body fluid identification. Methods An experimental system based on one-step reverse transcription polymerase chain reaction (RT-PCR) technology, capable of simultaneously detecting 12 RNAs, was constructed by screening specific mRNA markers, designing multiplex primers, and optimizing the reaction system. The kit was validated for its sensitivity, specificity, and repeatability using 275 samples from 5 common forensic body fluids and 10 casework samples. Positive controls were prepared using synthetic plasmids to complete the kit components. Results The kit effectively detected all 12 target RNAs within approximately 3 hours. It demonstrated high sensitivity, with a detection limit as low as 0.02 ng for certain sample types, along with excellent repeatability. Furthermore, the kit successfully identified specific mRNAs of individual components in mixed samples and exhibited strong performance in casework applications. Conclusion  This kit can be used to identify 5 common body fluid stains—peripheral blood, menstrual blood, semen, saliva, and vaginal secretions—and offers advantages such as simple operation, short processing time, and strong ability to identify mixed samples. It shows promising potential for practical forensic applications.

Key words: forensic genetics, mRNA, body fluid identification, one-step RT-PCR

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