"Pathotyping" Multiplex PCR Assay for Haemophilus parasuis: a Tool for Prediction of Virulence

J Clin Microbiol. 2017 Sep;55(9):2617-2628. doi: 10.1128/JCM.02464-16. Epub 2017 Jun 14.

Abstract

Haemophilus parasuis is a diverse bacterial species that is found in the upper respiratory tracts of pigs and can also cause Glässer's disease and pneumonia. A previous pangenome study of H. parasuis identified 48 genes that were associated with clinical disease. Here, we describe the development of a generalized linear model (termed a pathotyping model) to predict the potential virulence of isolates of H. parasuis based on a subset of 10 genes from the pangenome. A multiplex PCR (mPCR) was constructed based on these genes, the results of which were entered into the pathotyping model to yield a prediction of virulence. This new diagnostic mPCR was tested on 143 field isolates of H. parasuis that had previously been whole-genome sequenced and a further 84 isolates from the United Kingdom from cases of H. parasuis-related disease in pigs collected between 2013 and 2014. The combination of the mPCR and the pathotyping model predicted the virulence of an isolate with 78% accuracy for the original isolate collection and 90% for the additional isolate collection, providing an overall accuracy of 83% (81% sensitivity and 93% specificity) compared with that of the "current standard" of detailed clinical metadata. This new pathotyping assay has the potential to aid surveillance and disease control in addition to serotyping data.

Keywords: Haemophilus parasuis; molecular diagnostics; pathotyping; virulence factors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Genome / genetics
  • Haemophilus Infections / diagnosis*
  • Haemophilus Infections / microbiology
  • Haemophilus Infections / veterinary*
  • Haemophilus parasuis / genetics*
  • Haemophilus parasuis / isolation & purification
  • Haemophilus parasuis / pathogenicity*
  • Molecular Diagnostic Techniques / methods*
  • Multiplex Polymerase Chain Reaction
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / microbiology
  • Virulence / genetics