基于多种增菌方案的环境样本副溶血弧菌分离支持暴发事件溯源分析
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1.北京市顺义区疾病预防控制中心;2.中国疾病预防控制中心传染病预防控制所

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首都卫生发展科研专项


Isolation of Vibrio parahaemolyticus from environmental samples based on multiple bacterial enhancement schemes supports outbreak source tracking analysis
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Affiliation:

1.Shunyi District Center for Disease Control and Prevention, Beijing;2.National Institute for Communicable Disease Prevention and Control

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Capital’s Funds for Health Improvement and Research

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    摘要:

    目的 通过不同增菌方案分离环境样本中的副溶血弧菌,以支持一起暴发事件的溯源分析。方法 收集暴发事件流行病学资料、临床信息,采集多类型样本并使用多种增菌方案进行核酸检测和分离培养,对分离菌株进行全基因组测序并开展生物信息分析。结果 4件病例肛拭子和3件环境涂抹样本分离到7株副溶血弧菌,均为tdh+,血清型为O10:K4,基于菌株核心基因组单核苷酸多态性分析的结果显示,菌株间SNP差异介于0~5。全部环境涂抹样本使用含3%氯化钠的碱性蛋白胨水增菌后,副溶血弧菌的特异荧光PCR检测和培养实验结果均为阴性,环境涂抹样本E1、E2和E6使用脑心浸出液增菌后,副溶血弧菌的toxR和tdh荧光PCR检测结果均为toxR+/tdh+,其中E1和E2各靶基因Ct值>30,将脑心浸出液增菌后的E1和E2样本再次转种含3%氯化钠的碱性蛋白胨水二次增菌后,E1和E2各靶基因检测Ct值<30,且副溶血弧菌培养实验阳性。结论 本研究使用不同增菌方案分离环境样本中副溶血弧菌支持一起由该病原导致暴发事件调查溯源,环境涂抹样本中副溶血弧菌的有效增菌方法有待进一步深入研究。

    Abstract:

    Objective Different enrichment approaches were employed to isolate Vibrio parahaemolyticus (VP) from environmental samples, aiding in the traceability analysis of an outbreak. Methods Collected the epidemiological data and clinical information, and real time PCR and bacterial culture were performed in the samples. Whole genome sequencing and bioinformatics analysis were performed on the isolates in this outbreak. Results Seven strains of tdh+VP with O10:K4 serotype isolated from 4 patients and 3 environmental smear samples. The SNPs among the 7 strains ranged from 0 to 5 based on the analysis of single nucleotide polymorphisms in the core genome. All environmental smear samples were negative for VP by real time PCR and culture method based on 3%NaCl Alkaline Peptone Water (APW) enhancement. By brain heart infusion broth(BHI) enhancement, the real time PCR results showed that E1, E2 and E6 were toxR+/tdh+, and the Ct values of E1 and E2 were > 30 in all testd targets. The E1 and E2 samples cultured by BHI were re-cultured with 3%NaCl APW, and Ct values of E1 and E2 target genes were < 30, and VP culture was positive. Conclusion In this study, the isolation of VP in environmental samples was supported by different bacterial growth schemes to investigate and trace the source of an outbreak caused. The effective bacterial growth methods of VP in environmental smear samples need to be further studied.

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  • 收稿日期:2024-12-23
  • 最后修改日期:2025-04-13
  • 录用日期:2025-04-22
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