title: Development and evaluation of a multiplex real-time PCR assay for the detection and differentiation of Moraxella bovis, Moraxella bovoculi and Moraxella ovis in pure culture isolates and lacrimal swabs collected from conventionally raised cattle.
abstract: OBJECTIVE
  To develop a multiplex real-time PCR assay for the detection and differentiation of Moraxella bovis (M. bovis), M. bovoculi and M. ovis.
  
  
  RESULTS
  The multiplex real-time PCR assay was validated on three reference strains, 57 pure culture isolates and 45 lacrimal swab samples. All reference strains were identified correctly with no cross-reactions between species. Sequencing of 53 of the 57 culture isolates confirmed the results obtained with the multiplex real-time PCR, and the assay had 96·5% (55/57) concordance with a Moraxella spp. multiplex conventional PCR assay on the isolates. Among the lacrimal swab samples, the concordance between the multiplex real-time PCR and culture was 86·7% (39/45) for M. bovoculi and 75·6% (34/45) for M. bovis.
  
  
  CONCLUSIONS
  The multiplex real-time PCR assay is specific and sensitive and can be used directly on lacrimal swab samples.
  
  
  CONCLUSIONS
  The lack of a rapid, specific and sensitive detection method is a barrier for determining the roles of M. bovis, M. bovoculi and M. ovis in infectious bovine keratoconjunctivitis cases, and the developed PCR assay will contribute to improved understanding of the epidemiology and pathogenesis of these three Moraxella species.
authors.affiliation: Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
authors.forename: H G
authors.initials: HG
authors.lastname: Shen
authors.forename: S
authors.initials: S
authors.lastname: Gould
authors.forename: J
authors.initials: J
authors.lastname: Kinyon
authors.forename: T
authors.initials: T
authors.lastname: Opriessnig
authors.forename: A M
authors.initials: AM
authors.lastname: O'Connor
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics; D007765:Lacrimal Apparatus / Q000382:microbiology; D009016:Moraxella / Q000145:classification* / Q000235:genetics; D045828:Moraxellaceae Infections / Q000175:diagnosis* / Q000382:microbiology / Q000662:veterinary; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S