期刊
MOLECULAR AND CELLULAR PROBES
卷 55, 期 -, 页码 -出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.mcp.2020.101689
关键词
Multiplex PCR; Plesiomonas shigelloides; Vibrio
类别
资金
- National Science and Technology Major Project of China from the Ministry of Science and Technology of the People's Republic of China [2018ZX10712001-014, 2018ZX10305409-003-003]
- Wuxi Health Commission [T202017]
A multiplex PCR assay was developed to detect five pathogenic Vibrio species and Plesiomonas shigelloides, showing high specificity and sensitivity in various samples. It outperformed conventional cultivation methods in detection and can be applied for screening enrichment cultures and suspected colonies from different sources.
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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