A multiplex PCR assay for the detection of five human pathogenic Vibrio species and Plesiomonas

Hongxia Guan; Panpan Xue; Haijian Zhou; Dan Sha; Duochun Wang; He Gao; Jie Li; Baowei Diao; Hongqun Zhao; Biao Kan; Jingyun Zhang

doi:10.1016/j.mcp.2020.101689

A multiplex PCR assay for the detection of five human pathogenic Vibrio species and Plesiomonas

Mol Cell Probes. 2021 Feb:55:101689. doi: 10.1016/j.mcp.2020.101689. Epub 2020 Dec 15.

Authors

Hongxia Guan¹, Panpan Xue², Haijian Zhou², Dan Sha¹, Duochun Wang², He Gao², Jie Li², Baowei Diao², Hongqun Zhao², Biao Kan³, Jingyun Zhang⁴

Affiliations

¹ Wuxi Center for Disease Control and Prevention, Jiangsu, 214023, China.
² State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China.
³ State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China; Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, 310003, China.
⁴ State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China. Electronic address: zhangjingyun@icdc.cn.

PMID: 33338586
DOI: 10.1016/j.mcp.2020.101689

Abstract

A multiplex PCR (mPCR) assay was established to detect five pathogenic Vibrio species and Plesiomonas shigelloides. Twelve genes were included: ompW, ctxA, rfbN, and wbfR from V. cholerae; tl, tdh, and trh from V. parahaemolyticus; toxR and vmhA from V. mimicus; toxR from V. fluvialis; vvhA from V. vulnificus; and the 23S rRNA gene from P. shigelloides. The specificity of the mPCR assay was 100% for the detection of 136 strains and the limits of detection (LoD) were 12.5-50 pg/reaction. The assay exhibited higher sensitivity than cultivation methods in the detection of APW cultures of 113 diarrhea samples. In the analysis of 369 suspected Vibrio populations from estuarine water samples, the specificity of the mPCR for V. cholerae and V. parahaemolyticus was 100% for both, while the sensitivities were 100% and 96.1%, respectively. The assay can be applied to screen enrichment cultures and suspected colonies from environmental and clinical samples.

Keywords: Multiplex PCR; Plesiomonas shigelloides; Vibrio.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Electrophoresis, Capillary
Estuaries
Humans
Multiplex Polymerase Chain Reaction / methods*
Plesiomonas / genetics*
Plesiomonas / isolation & purification*
Sensitivity and Specificity
Vibrio / genetics*
Vibrio / isolation & purification*
Water Microbiology