PLoS ONE (Jan 2014)

Evaluation of ERIC-PCR as genotyping method for Corynebacterium pseudotuberculosis isolates.

  • Elaine M S Dorneles,
  • Jordana A Santana,
  • Dayana Ribeiro,
  • Fernanda Alves Dorella,
  • Alessandro S Guimarães,
  • Mohamed S Moawad,
  • Salah A Selim,
  • Ana Luiza M Garaldi,
  • Anderson Miyoshi,
  • Márcio G Ribeiro,
  • Aurora M G Gouveia,
  • Vasco Azevedo,
  • Marcos B Heinemann,
  • Andrey P Lage

DOI
https://doi.org/10.1371/journal.pone.0098758
Journal volume & issue
Vol. 9, no. 6
p. e98758

Abstract

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The aim of this study was to evaluate the Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) as a tool for molecular typing of C. pseudotuberculosis isolates from eight different hosts in twelve countries. Ninety-nine C. pseudotuberculosis field strains, one type strain (ATCC 19410T) and one vaccine strain (1002) were fingerprinted using the ERIC-1R and ERIC-2 primers, and the ERIC-1R+ERIC-2 primer pair. Twenty-nine different genotypes were generated by ERIC 1-PCR, 28 by ERIC 2-PCR and 35 by ERIC 1+2-PCR. The discriminatory index calculated for ERIC 1, ERIC 2, and ERIC 1+2-PCR was 0.89, 0.86, and 0.92, respectively. Epidemiological concordance was established for all ERIC-PCR assays. ERIC 1+2-PCR was defined as the best method based on suitability of the amplification patterns and discriminatory index. Minimal spanning tree for ERIC 1+2-PCR revealed three major clonal complexes and clustering around nitrate-positive (biovar Equi) and nitrate-negative (biovar Ovis) strains. Therefore, ERIC 1+2-PCR proved to be the best technique evaluated in this study for genotyping C. pseudotuberculosis strains, due to its usefulness for molecular epidemiology investigations.