The screening and validation process of miR-223-3p for saliva identification

Huixiang Chen; Sheng Hu; Rui Yang; Shuxiao Hu; Qianwei Yao; Yixia Zhao; Jie Lian; Anquan Ji; Yang Cao; Qifan Sun

doi:10.1016/j.legalmed.2023.102312

The screening and validation process of miR-223-3p for saliva identification

Leg Med (Tokyo). 2023 Nov:65:102312. doi: 10.1016/j.legalmed.2023.102312. Epub 2023 Aug 9.

Authors

Huixiang Chen¹, Sheng Hu², Rui Yang³, Shuxiao Hu⁴, Qianwei Yao¹, Yixia Zhao², Jie Lian⁵, Anquan Ji⁶, Yang Cao⁷, Qifan Sun⁸

Affiliations

¹ MPS's Key Laboratory of Forensic Genetics, National Engineering Laboratory for Crime Scene Evidence Investigation and Examination, Institute of Forensic Science, Ministry of Public Security (MPS), Beijing 100038, China; Faculty of Forensic Sciences, Shanxi Medical University, Taiyuan 030001, Shanxi, China.
² MPS's Key Laboratory of Forensic Genetics, National Engineering Laboratory for Crime Scene Evidence Investigation and Examination, Institute of Forensic Science, Ministry of Public Security (MPS), Beijing 100038, China.
³ Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, China.
⁴ MPS's Key Laboratory of Forensic Genetics, National Engineering Laboratory for Crime Scene Evidence Investigation and Examination, Institute of Forensic Science, Ministry of Public Security (MPS), Beijing 100038, China; School of Investigation, People's Public Security University of China, Beijing 100038, China.
⁵ School of Investigation, People's Public Security University of China, Beijing 100038, China.
⁶ MPS's Key Laboratory of Forensic Genetics, National Engineering Laboratory for Crime Scene Evidence Investigation and Examination, Institute of Forensic Science, Ministry of Public Security (MPS), Beijing 100038, China. Electronic address: aqjdna@163.com.
⁷ Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, China. Electronic address: cao@scu.edu.cn.
⁸ MPS's Key Laboratory of Forensic Genetics, National Engineering Laboratory for Crime Scene Evidence Investigation and Examination, Institute of Forensic Science, Ministry of Public Security (MPS), Beijing 100038, China. Electronic address: sunqifan@cifs.gov.cn.

PMID: 37603982
DOI: 10.1016/j.legalmed.2023.102312

Abstract

More accurate identification of the types of body fluids left at a crime scene is indispensable for improving the judicial chain of evidence. MicroRNAs (miRNAs) have become recognized as ideal molecular markers for the identification of body fluids in forensic science due to their short length, stability and high tissue specificity. In this study, small RNA sequencing was performed on 20 samples of five types of body fluids (peripheral blood, menstrual blood, saliva, semen, and vaginal secretions) with the BGISEQ-500 sequencing platform, and the specific miRNA markers of saliva and vaginal secretions were screened by bioinformatics methods, including differential expression analysis and significant enrichment analysis. Through RT-qPCR validation of 169 samples, we confirmed that miR-223-3p can be used as a saliva-specific marker. In addition, we considered miR-223-3p in combination with four other miRNA molecules (miR-451a, miR-891a-5p, miR-144-5p, miR-203a-3p) that had been previously screened and verified in our laboratory, and seven body fluid prediction models based on machine learning algorithms were constructed and verified. The results showed that a kernel density estimation (KDE) model based on the five miRNA markers for body fluid identification could achieve 100% accuracy in the samples tested in the present study.

Keywords: Bioinformatics; Body fluid identification; Forensic science; High-throughput sequencing; RT-qPCR; miRNA.

MeSH terms

Biomarkers / metabolism
Body Fluids* / chemistry
Female
Forensic Genetics / methods
Humans
MicroRNAs* / analysis
Saliva

Substances

MicroRNAs
Biomarkers
MIRN223 microRNA, human