Departamento de Biología Funcional (Área de Microbiología), Universidad de Oviedo, 33006-Oviedo, Asturias, Spain.
J Antimicrob Chemother. 2011 Mar;66(3):543-9. doi: 10.1093/jac/dkq481. Epub 2010 Dec 21.
To broaden knowledge of the molecular bases and genetics of multidrug resistance in clinical isolates of Salmonella enterica serotype 4,5,12:i:- belonging to the Spanish clone.
The relatedness of the isolates was determined by phage typing and XbaI-PFGE. Resistance genes, integrons and transposable elements were identified by PCR amplification and sequencing. Plasmids were characterized by alkaline lysis, S1-PFGE, conjugation, replicon typing and Southern blot hybridization.
The isolates were closely related and resistant to five to seven antimicrobials (ampicillin, chloramphenicol, gentamicin, streptomycin/spectinomycin, sulphonamides, trimethoprim and tetracycline, arranged in different combinations). Most of the responsible genes were provided by a conventional class 1 integron with the dfrA12-orfF-aadA2 variable region, an atypical class 1 integron containing sul3 next to the estX-psp-aadA2-cmlA1-aadA1 variable region and a truncated Tn1721 transposon carrying tet(A). A defective Tn21 with the mer operon and ISVsa3 associated with sul2 were also detected. All resistance genes and mobile genetic elements were located on large, non-conjugative and highly variable plasmids carrying one (A/C) or two (A/C and N) replicons, as well as virulence genes of pSLT.
IncA/C plasmids are responsible for multidrug resistance in an increasing number of relevant human and animal bacterial pathogens, and hence are regarded as an important threat to public health. Those found in the Spanish clone of Salmonella 4,5,12:i:- constitute a relevant example of short-term evolution, and could have been involved in the successful adaptation of this pathogen.
拓宽对属于西班牙克隆的沙门氏菌血清型 4、5、12:i:-临床分离株中多药耐药的分子基础和遗传学的认识。
通过噬菌体分型和 XbaI-PFGE 确定分离株的相关性。通过 PCR 扩增和测序鉴定耐药基因、整合子和可移动元件。通过碱性裂解、S1-PFGE、接合、复制子分型和 Southern 印迹杂交来表征质粒。
分离株密切相关,对五种至七种抗菌药物(氨苄西林、氯霉素、庆大霉素、链霉素/壮观霉素、磺胺类药物、甲氧苄啶和四环素,以不同组合排列)耐药。大多数相关基因由一个常规的 1 类整合子提供,带有 dfrA12-orfF-aadA2 可变区,一个含有 sul3 的非典型 1 类整合子,紧邻 estX-psp-aadA2-cmlA1-aadA1 可变区,以及一个携带 tet(A)的截断 Tn1721 转座子。还检测到带有 mer 操纵子和与 sul2 相关的 ISVsa3 的缺陷型 Tn21。所有耐药基因和可移动遗传元件均位于携带一个(A/C)或两个(A/C 和 N)复制子的大型非接合和高度可变的质粒上,以及 pSLT 的毒力基因。
IncA/C 质粒是越来越多相关人类和动物细菌病原体多药耐药的原因,因此被认为是对公共健康的一个重要威胁。在沙门氏菌 4、5、12:i:-的西班牙克隆中发现的 IncA/C 质粒构成了短期进化的一个重要例子,可能参与了该病原体的成功适应。