Citation

Xiao C, Xu Q, Li H, Zhu X, Yu Z, Zhao J, Li Y, Liu H, Shi H, Liu C. Electrophoresis 2021; ePub(ePub): ePub.

Copyright

(Copyright © 2021, Wiley-VCH)

DOI

10.1002/elps.202000265

PMID

unavailable

Abstract

In recent years, the DNA detection of drowning-related diatoms, cyanobacteria and aeromonas has gradually attracted interest from forensic scientists. In this study, we described the validation and application of a novel multiplex PCR system. This system integrated 12 fluorescently labelled primers designed to amplify specific genes of diatoms, cyanobacteria and aeromonas. The specificity studies demonstrated that this multiplex PCR system could detect 9 species of diatom, 7 species of cyanobacteria and 5 species of aeromonas, all of which were drowning-related and widely distributed in various water circumstance of southern China. The sensitivity studies indicated that the limit concentration of template DNA was 0.0125 ng. Besides, this multiplex PCR system had good performance in sizing precision and stability, but it is not suitable for degraded DNA samples. The application into forensic casework showed that all the tissue samples from ten non-drowning cases showed negative results, and the positive rates of lung, liver, kidney, and water samples from thirty drowning bodies were 100%, 86.7%, 90% and 100%, respectively. Combined with results of diatom tests of MD-VF-Auto SEM method, this multiplex PCR system could help rule out non-drowning bodies and provide extra evidences to support drowning diagnosis, especially for those cases with few diatoms were observed. It is expected that this multiplex PCR system has great potential for forensic drowning diagnosis. This article is protected by copyright. All rights reserved.

Language: en

Keywords

Diatom test; Drowning diagnosis; Capillary Electrophoresis; Multiplex PCR system