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. 2018 Aug 15;13(8):e0201428.
doi: 10.1371/journal.pone.0201428. eCollection 2018.

Genome-driven Evaluation and Redesign of PCR Tools for Improving the Detection of Virulence-Associated Genes in Aeromonads

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Free PMC article

Genome-driven Evaluation and Redesign of PCR Tools for Improving the Detection of Virulence-Associated Genes in Aeromonads

Emilie Talagrand-Reboul et al. PLoS One. .
Free PMC article

Abstract

Many virulence factors have been described for opportunistic pathogens within the genus Aeromonas. Polymerase Chain Reactions (PCRs) are commonly used in population studies of aeromonads to detect virulence-associated genes in order to better understand the epidemiology and emergence of Aeromonas from the environment to host, but their performances have never been thoroughly evaluated. We aimed to determine diagnostic sensitivity and specificity of PCR assays for the detection of virulence-associated genes in a collection of Aeromonas isolates representative for the genetic diversity in the genus. Thirty-nine Aeromonas strains belonging to 27 recognized species were screened by published PCR assays for virulence-associated genes (act, aerA, aexT, alt, ascFG, ascV, ast, lafA, lip, ser, stx1, stx2A). In parallel, homologues of the 12 putative virulence genes were searched from the genomes of the 39 strains. Of the 12 published PCR assays for virulence factors, the comparison of PCR results and genome analysis estimated diagnostic sensitivities ranging from 34% to 100% and diagnostic specificities ranged from 71% to 100% depending upon the gene. To improve the detection of virulence-associated genes in aeromonads, we have designed new primer pairs for aerA/act, ser, lafA, ascFG and ascV, which showed excellent diagnostic sensitivity and specificity. Altogether, the analysis of high quality genomic data, which are more and more easy to obtain, provides significant improvements in the genetic detection of virulence factors in bacterial strains.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Flowchart for the literature analysis.
The automatic Pubmed query using « Aeromonas », « PCR » and « virulence » as keywords generated 126 articles (October 27, 2016), among which 46 corresponded to the entry criteria. The reading of the relevant references quoted in these 46 selected articles led to the manual inclusion of 15 additional articles.

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Grant support

This work was supported by the Association des Biologistes de l’Ouest (ABO) and the Association pour la recherche et le développement en microbiologie et pharmacie (ADEREMPHA) to ETR. Part of this research was supported by USDA and ARS agreement 58-1930-4-002 to JG.
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