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

被引：15
|
作者：
Guan, Hongxia [1 ]
Xue, Panpan [2 ]
Zhou, Haijian [2 ]
Sha, Dan [1 ]
Wang, Duochun [2 ]
Gao, He [2 ]
Li, Jie [2 ]
Diao, Baowei [2 ]
Zhao, Hongqun [2 ]
Kan, Biao [2 ,3 ]
Zhang, Jingyun [2 ]
机构：
[1] Wuxi Ctr Dis Control & Prevent, Wuxi 214023, Jiangsu, Peoples R China
[2] Chinese Ctr Dis Control & Prevent, Natl Inst Communicable Dis Control & Prevent, State Key Lab Infect Dis Prevent & Control, 155 Changbai Rd, Beijing 102206, Peoples R China
[3] Collaborat Innovat Ctr Diag & Treatment Infect Di, Hangzhou 310003, Peoples R China
来源：
MOLECULAR AND CELLULAR PROBES | 2021年 / 55卷
关键词：
Multiplex PCR; Plesiomonas shigelloides; Vibrio;
D O I：
10.1016/j.mcp.2020.101689
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
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页数：6
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