一种用于检测五种人类病原体弧菌和类志贺邻单胞菌的多重 PCR 检测方法。
A multiplex PCR assay for the detection of five human pathogenic Vibrio species and Plesiomonas.
机构信息
Wuxi Center for Disease Control and Prevention, Jiangsu, 214023, China.
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.
出版信息
Mol Cell Probes. 2021 Feb;55:101689. doi: 10.1016/j.mcp.2020.101689. Epub 2020 Dec 15.
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.
建立了一种多重 PCR（mPCR）检测方法，用于检测五种致病性弧菌和类志贺邻单胞菌。该方法包含 12 个基因：霍乱弧菌的 ompW、ctxA、rfbN 和 wbfR；副溶血弧菌的 tl、tdh 和 trh；类河弧菌的 toxR 和 vmhA；弗氏弧菌的 toxR；创伤弧菌的 vvhA；以及类志贺邻单胞菌的 23S rRNA 基因。mPCR 检测方法对 136 株菌株的检测特异性为 100%，检测限（LoD）为 12.5-50 pg/反应。与培养方法相比，该方法在检测 113 份腹泻样本的 APW 培养物时具有更高的灵敏度。在对来自河口水样的 369 个疑似弧菌种群的分析中，mPCR 对霍乱弧菌和副溶血弧菌的特异性均为 100%，灵敏度分别为 100%和 96.1%。该方法可应用于筛选环境和临床样本中的富集培养物和疑似菌落。
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