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肠沙门氏菌肠炎亚种中毒力和抗微生物耐药质粒 pOU7519 的遗传分析。

Genetic analysis of virulence and antimicrobial-resistant plasmid pOU7519 in Salmonella enterica serovar Choleraesuis.

机构信息

Molecular Infectious Disease Research Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan.

Molecular Infectious Disease Research Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan; Department of Laboratory Medicine, Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Taoyuan, Taiwan.

出版信息

J Microbiol Immunol Infect. 2020 Feb;53(1):49-59. doi: 10.1016/j.jmii.2017.11.004. Epub 2017 Dec 14.

Abstract

BACKGROUND

Zoonotic Salmonella enterica serovar Choleraesuis (S. Choleraesuis), causing paratyphoid in pigs and bacteremia in humans, commonly carry a virulence plasmid and sometimes a separate antimicrobial-resistant plasmid or merging together. This study aimed to analyze the likely mechanism of how to form a virulence-resistance chimera of plasmid in S. Choleraesuis.

METHODS

Whole plasmid sequence of pOU7519 in S. Choleraesuis strain OU7519 was determined using shotgun cloning and sequencing. Sequence annotation and comparison were performed to determine the sequence responsible for the formation of a chimeric virulence-resistance pOU7519. Other chimeric plasmids among the collected strains of S. Choleraesuis were also confirmed.

RESULTS

The sequence of pOU719, 127,212 bp long, was identified to be a chimera of the virulence plasmid pSCV50 and a multidrug-resistant plasmid pSC138 that have been found in S. Choleraesuis strain SC-B67. The pOU7519 is a conjugative plasmid carrying various mobile DNAs, including prophages, insertion sequences, integrons and transposons, especially a Tn6088-like transposon. By dissecting the junction site of the pSCV50-pSC138 chimera in pOU7519, defective sequences at integrase gene scv50 (int) and its attachment site (att) were found, and that likely resulted in a stable chimera plasmid due to the failure of excision from the pSCV50-pSC138 chimera. Similar structure of chimera was also found in other large plasmids.

CONCLUSION

The deletion of both the int and att sties could likely block chimera excision, and result in an irreversible, stable pSCV50-pSC138 chimera. The emergence of conjugative virulence and antimicrobial-resistant plasmids in S. Choleraesuis could pose a threat to health public.

摘要

背景

引起猪副伤寒和人类菌血症的人畜共患沙门氏菌肠炎血清型(S. Choleraesuis)通常携带毒力质粒,有时还携带单独的抗微生物耐药质粒或融合在一起。本研究旨在分析 S. Choleraesuis 中质粒毒力-耐药嵌合体形成的可能机制。

方法

使用鸟枪法克隆和测序确定 S. Choleraesuis 菌株 OU7519 中 pOU7519 的全质粒序列。进行序列注释和比较,以确定形成嵌合毒力-耐药 pOU7519 的序列。还确认了收集的 S. Choleraesuis 菌株中的其他嵌合质粒。

结果

鉴定出长 127,212 bp 的 pOU719 序列是已在 S. Choleraesuis 菌株 SC-B67 中发现的毒力质粒 pSCV50 和多药耐药质粒 pSC138 的嵌合体。pOU7519 是一种带有各种移动 DNA 的可接合质粒,包括噬菌体、插入序列、整合子和转座子,特别是 Tn6088 样转座子。通过剖析 pOU7519 中 pSCV50-pSC138 嵌合体的连接点,发现了 scv50(int)整合酶基因及其附着位点(att)的缺陷序列,由于无法从 pSCV50-pSC138 嵌合体中切除,这可能导致稳定的嵌合体质粒。在其他大质粒中也发现了类似的嵌合体结构。

结论

int 和 att 位点的缺失可能会阻止嵌合体的切除,从而导致不可逆的稳定的 pSCV50-pSC138 嵌合体。S. Choleraesuis 中出现的可接合毒力和抗微生物耐药质粒可能对公共卫生健康构成威胁。

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