title: Evaluation of a multiplex PCR to identify and serotype Actinobacillus pleuropneumoniae serovars 1, 5, 7, 12 and 15.
abstract: The aim of this study was to validate a multiplex PCR for the species identification and serotyping of Actinobacillus pleuropneumoniae serovars 1, 5, 7, 12 and 15. All 15 reference strains and 411 field isolates (394 from Australia, 11 from Indonesia, five from Mexico and one from New Zealand) of A. pleuropneumoniae were tested with the multiplex PCR. The specificity of this multiplex PCR was validated on 26 non-A. pleuropneumoniae species. The multiplex PCR gave the expected results with all 15 serovar reference strains and agreed with conventional serotyping for all field isolates from serovars 1 (n = 46), 5 (n = 81), 7 (n = 80), 12 (n = 16) and serovar 15 (n = 117). In addition, a species-specific product was amplified in the multiplex PCR with all 411 A. pleuropneumoniae field isolates. Of 25 nontypeable field isolates only two did not yield a serovar-specific band in the multiplex PCR. This multiplex PCR for serovars 1, 5, 7, 12 and 15 is species specific and capable of serotyping isolates from diverse locations. Significance and impact of the study: A multiplex PCR that can recognize serovars 1, 5, 7, 12 and 15 of A. pleuropneumoniae was developed and validated. This novel diagnostic tool will enable frontline laboratories to provide key information (the serovar) to guide targeted prevention and control programmes for porcine pleuropneumonia, a serious economic disease of pigs. The previous technology, traditional serotyping, is typically provided by specialized reference laboratories, limiting the capacity to respond to this key disease.
authors.affiliation: Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, Dutton Park, Qld, Australia.
authors.forename: C
authors.initials: C
authors.lastname: Turni
authors.forename: R
authors.initials: R
authors.lastname: Singh
authors.forename: M A
authors.initials: MA
authors.lastname: Schembri
authors.forename: P J
authors.initials: PJ
authors.lastname: Blackall
mesh_terms: D000189:Actinobacillus Infections; D016977:Actinobacillus pleuropneumoniae / Q000145:classification* / Q000302:isolation & purification*; D001315:Australia; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009174:Mycoplasma / Q000235:genetics; D065288:Serogroup; D012703:Serotyping / Q000379:methods*; D013045:Species Specificity
keywords: Actinobacillus pleuropneumoniae; identification; multiplex PCR; serotyping; validation