Silva Claudia, Betancor Laura, García Coralith, Astocondor Lizeth, Hinostroza Noemí, Bisio Julieta, Rivera Javier, Perezgasga Lucía, Pérez Escanda Victoria, Yim Lucía, Jacobs Jan, García-Del Portillo Francisco, Chabalgoity José A, Puente José L
Departamento de Microbiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, Mexico.
Instituto de Higiene, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
PLoS One. 2017 Dec 21;12(12):e0189946. doi: 10.1371/journal.pone.0189946. eCollection 2017.
In this study, different molecular typing tools were applied to characterize 95 Salmonella enterica blood isolates collected between 2008 and 2013 from patients at nine public hospitals in Lima, Peru. Combined results of multiplex PCR serotyping, two- and seven-loci multilocus sequence typing (MLST) schemes, serotyping, IS200 amplification and RAPD fingerprints, showed that these infections were caused by eight different serovars: Enteritidis, Typhimurium, Typhi, Choleraesuis, Dublin, Paratyphi A, Paratyphi B and Infantis. Among these, Enteritidis, Typhimurium and Typhi were the most prevalent, representing 45, 36 and 11% of the isolates, respectively. Most isolates (74%) were not resistant to ten primarily used antimicrobial drugs; however, 37% of the strains showed intermediate susceptibility to ciprofloxacin (ISC). Antimicrobial resistance integrons were carried by one Dublin (dfra1 and aadA1) and two Infantis (aadA1) isolates. The two Infantis isolates were multidrug resistant and harbored a large megaplasmid. Amplification of spvC and spvRA regions showed that all Enteritidis (n = 42), Typhimurium (n = 34), Choleraesuis (n = 3) and Dublin (n = 1) isolates carried the Salmonella virulence plasmid (pSV). We conclude that the classic serotyping method can be substituted by the multiplex PCR and, when necessary, sequencing of only one or two loci of the MLST scheme is a valuable tool to confirm the results. The effectiveness and feasibility of different typing tools is discussed.
在本研究中,应用了不同的分子分型工具来鉴定2008年至2013年间从秘鲁利马九家公立医院的患者中收集的95株肠炎沙门氏菌血液分离株。多重PCR血清分型、双位点和七位点多位点序列分型(MLST)方案、血清分型、IS200扩增和RAPD指纹图谱的综合结果表明,这些感染是由八种不同的血清型引起的:肠炎型、鼠伤寒型、伤寒型、猪霍乱型、都柏林型、甲型副伤寒型、乙型副伤寒型和婴儿型。其中,肠炎型、鼠伤寒型和伤寒型最为常见,分别占分离株的45%、36%和11%。大多数分离株(74%)对十种主要使用的抗菌药物不耐药;然而,37%的菌株对环丙沙星表现出中度敏感性(ISC)。一株都柏林分离株(携带dfra1和aadA1)和两株婴儿分离株(携带aadA1)携带抗菌耐药整合子。这两株婴儿分离株具有多重耐药性,并携带一个大的巨型质粒。spvC和spvRA区域的扩增表明,所有肠炎型(n = 42)、鼠伤寒型(n = 34)、猪霍乱型(n = 3)和都柏林型(n = 1)分离株都携带沙门氏菌毒力质粒(pSV)。我们得出结论,多重PCR可以替代经典血清分型方法,并且在必要时,MLST方案中仅一个或两个位点的测序是确认结果的有价值工具。本文还讨论了不同分型工具的有效性和可行性。