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dc.contributor.authorStringer, Oliver W
dc.contributor.authorLi, Yanwen
dc.contributor.authorBossé, Janine T
dc.contributor.authorForrest, Matthew S
dc.contributor.authorHernandez-Garcia, Juan
dc.contributor.authorTucker, Alexander W
dc.contributor.authorNunes, Tiago
dc.contributor.authorCosta, Francisco
dc.contributor.authorMortensen, Preben
dc.contributor.authorVelazquez, Eduardo
dc.contributor.authorPenny, Paul
dc.contributor.authorRodriguez-Manzano, Jesus
dc.contributor.authorGeorgiou, Pantelis
dc.contributor.authorLangford, Paul R
dc.description.abstractActinobacillus pleuropneumoniae (APP) is the causative agent of porcine pleuropneumonia, resulting in high economic impact worldwide. There are currently 19 known serovars of APP, with different ones being predominant in specific geographic regions. Outbreaks of pleuropneumonia, characterized by sudden respiratory difficulties and high mortality, can occur when infected pigs are brought into naïve herds, or by those carrying different serovars. Good biosecurity measures include regular diagnostic testing for surveillance purposes. Current gold standard diagnostic techniques lack sensitivity (bacterial culture), require expensive thermocycling machinery (PCR) and are time consuming (culture and PCR). Here we describe the development of an isothermal point-of-care diagnostic test - utilizing recombinase polymerase amplification (RPA) for the detection of APP, targeting the species-specific apxIVA gene. Our APP-RPA diagnostic test achieved a sensitivity of 10 copies/μL using a strain of APP serovar 8, which is the most prevalent serovar in the UK. Additionally, our APP-RPA assay achieved a clinical sensitivity and specificity of 84.3 and 100%, respectively, across 61 extracted clinical samples obtained from farms located in England and Portugal. Using a small subset (n = 14) of the lung tissue samples, we achieved a clinical sensitivity and specificity of 76.9 and 100%, respectively) using lung imprints made on FTA cards tested directly in the APP-RPA reaction. Our results demonstrate that our APP-RPA assay enables a suitable rapid and sensitive screening tool for this important veterinary pathogen.
dc.publisherFrontiers Media SA
dc.rightsAttribution 4.0 International
dc.sourcenlmid: 101666658
dc.sourceessn: 2297-1769
dc.subjectA. pleuropneumoniae
dc.subjectFTA® card
dc.subjectRPA (recombinase polymerase amplification)
dc.subjectpoint-of-care (POC)
dc.titleRapid Detection of Actinobacillus pleuropneumoniae From Clinical Samples Using Recombinase Polymerase Amplification.
prism.publicationNameFront Vet Sci
dc.contributor.orcidBossé, Janine T [0000-0002-8491-4779]
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/M011178/1, BB/S005897/1, BB/S002103/1)

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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International