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ARS Home » Northeast Area » Leetown, West Virginia » Cool and Cold Water Aquaculture Research » Research » Publications at this Location » Publication #365575
Research Project: Integrated Research to Improve On-Farm Animal Health in Salmonid Aquaculture

Location: Cool and Cold Water Aquaculture Research

Title: Genome-driven evaluation and redesign of PCR tools for improving the detection of virulence-associated genes in aeromonads

Author
item TALAGRAND-REBOUL, EMILIE - University Of Montpellier
item LATIF-EUGENIN, FADUA - University Rovira I Virgili
item BEAZ-HIDALGO, ROXANA - University Rovira I Virgili
item COLSTON, SOPHIE - University Of Connecticut
item FIGUERAS, MARIA-JOSE - University Rovira I Virgili
item GRAF, JOERG - University Of Connecticut
item JUMAS-BILAK, ESTELLE - University Of Montpellier
item LAMY, BRIGITTE - University Of Montpellier

Submitted to: PLOS ONE
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/7/2018
Publication Date: 8/15/2018
Citation: Talagrand-Reboul, E., Latif-Eugenin, F., Beaz-Hidalgo, R., Colston, S., Figueras, M., Graf, J., Jumas-Bilak, E., Lamy, B. 2018. Genome-driven evaluation and redesign of PCR tools for improving the detection of virulence-associated genes in aeromonads. PLoS One. 13(8):e0201428. https://doi.org/10.1371/journal.pone.0201428.
DOI: https://doi.org/10.1371/journal.pone.0201428

Interpretive Summary: A number of fish pathogens are Aeromonas species. One important aspect that determines how pathogenic different strains are is the complement of virulence factors they carry. In this manuscript PCR assays for 12 genes important in causing disease were tested on 39 Aeromonas strains belonging to 27 species. Five of the primers were improved to work better across the wide range of species using genomic data. This paper is an important step forward in identifying virulence factors in Aeromonas

Technical 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.