SYBR Green real-time qPCR method: Diagnose drowning more rapidly and accurately.

Zhonghao Yu, Quyi Xu, Cheng Xiao, Huan Li, Weibin Wu, Weian Du, Jian Zhao, Hong Liu, Huijun Wang, Chao Liu

Journal: Forensic science international 2021;321():110720

PMID: 33639416

Full Text on Elsevier Science View on PubMed

Abstract

In the field of drowning research, the method of diatom morphology has been most applied to determine whether the cause of death is drowning. However, the characteristics of complex operation, high level of professional knowledge drive us to propose a new method. Here, based on the common phytoplankton in water(such as diatoms and Aeromonas), aiming at the rbcL, 23 S, NIES, rPOD, Hly and preprotoxin aerolysin gene, we designed 6 pairs of specific primers and applied SYBR Green real-time qPCR(RT-qPCR) method to detect phytoplankton in the Pearl River Basin of Guangdong Province, China, so as to achieve the purpose of diagnosing drowning. After the experimental verification of the corresponding algae species and the standard strains of bacteria, as well as the verification of tissue samples (lung, liver and kidney) of 56 cases( 40 drowning cases and 16 non-drowning cases), we found that these primers were of great accuracy and tedious laboratory work of diatom test was reduced. Based on the advantages of high throughput, short period and high sensitivity, this RT-qPCR method is expected to diagnose drowning more rapidly and accurately.

Address: School of Forensic Medicine, Southern Medical University, Guangzhou 510515, China.; Guangzhou Forensic Science Institute, Guangdong Province Key Laboratory of Forensic Genetics, Guangzhou 510030, China.; Department of Forensic Medicine, Faculty of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China.; HOMY GeneTech Incorporation, Foshan 518000, China.; School of Forensic Medicine, Southern Medical University, Guangzhou 510515, China. Electronic address: [email protected].; School of Forensic Medicine, Southern Medical University, Guangzhou 510515, China; Guangzhou Forensic Science Institute, Guangdong Province Key Laboratory of Forensic Genetics, Guangzhou 510030, China. Electronic address: [email protected].

DOI: 10.1016/j.forsciint.2021.110720

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MeSH Terms: Aeromonas, Benzothiazoles, Cyanobacteria, DNA Primers, Diamines, Diatoms, Drowning, Fluorescent Dyes, Humans, Kidney, Liver, Lung, Quinolines, Real-Time Polymerase Chain Reaction