副溶血弧菌在乳鼠肠道内竞争性定殖实验方法的建立及应用[J]. 疾病监测, 2015, 30(12): 1050-1055. DOI: 10.3784/j.issn.1003-9961.2015.12.015
引用本文: 副溶血弧菌在乳鼠肠道内竞争性定殖实验方法的建立及应用[J]. 疾病监测, 2015, 30(12): 1050-1055. DOI: 10.3784/j.issn.1003-9961.2015.12.015
Establishment and application of assays for evaluating competitive colonization of Vibrio parahaemolyticus in infant mouse intestinal track[J]. Disease Surveillance, 2015, 30(12): 1050-1055. DOI: 10.3784/j.issn.1003-9961.2015.12.015
Citation: Establishment and application of assays for evaluating competitive colonization of Vibrio parahaemolyticus in infant mouse intestinal track[J]. Disease Surveillance, 2015, 30(12): 1050-1055. DOI: 10.3784/j.issn.1003-9961.2015.12.015

副溶血弧菌在乳鼠肠道内竞争性定殖实验方法的建立及应用

Establishment and application of assays for evaluating competitive colonization of Vibrio parahaemolyticus in infant mouse intestinal track

  • 摘要: 目的 建立评价副溶血弧菌在乳鼠肠道内定殖能力的实验方法,为研究副溶血弧菌的致病机制奠定基础。方法 构建带有霍乱弧菌lacZ基因的副溶血弧菌临床分离株RIMD2210633(WT)的LacZ菌株(WT-lacZ);比较WT菌株与WT-lacZ菌株以及WT-lacZ菌株与环境分离株VP1907在CD-1乳鼠(5日龄)肠道定殖能力的差别。结果 成功构建X-Gal LB平板上显蓝色的副溶血弧菌LacZ菌株。WT-lacZ菌株和WT菌株在乳鼠肠道内定殖能力无差别,这表明可以用WT-lacZ菌株代替WT菌株进行后续的实验;VP1907与WT-lacZ菌株在乳鼠肠道内定殖的竞争系数在0.1左右,表明WT菌株的定殖能力明显高于VP1907菌株。结论 建立了评价副溶血弧菌肠道定殖能力的乳鼠模型,并应用该模型进行了临床分离株与环境分离株肠道定殖能力的比较分析。

     

    Abstract: Objective To establish an infant mouse model to evaluate the competitive colonization of Vibrio parahaemolyticus in mouse intestinal track and provide evidence for the research of pathogenicity of V. parahaemolyticus. Methods The -galactosidase gene lacZ of Vibrio cholerae was amplified and cloned into the V. parahaemolyticus RIMD2210633(WT) genome to construct WT- lacZ strain. Then, the colonization ability of the WT-lacZ strain in 5-day-old CD-1 suckling mice intestine was compared with that of the WT strain and an environment-isolated VP1907 strain, respectively. Results The colony of WT-lacZ strain was blue on the X-Gal LB agar plate. The WT-lacZ strain showed no significant change in colonization compared with WT strain, suggesting that the WT-lacZ strain can substitute the WT strain in colonization competition study in vivo. The result of the in vivo competition assay showed that the competitive index between the WT-lacZ strain and VP1907 strain was 0.1 approximately, indicating significantly higher colonization of the WT strain than that of the VP1907 strain. Conclusion A new infant mouse model using RIMD2210633 strain was successfully established for evaluating the intestinal competitive colonization of V. parahaemolyticus and used to compare the competitive colonization between highly virulent strains and non-virulent strains isolated in environment.

     

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