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dc.contributor.authorHowell, Kateen
dc.contributor.authorMaskell, Sarahen
dc.contributor.authorWang, Jinhongen
dc.contributor.authorHernandez-Garcia, Juanen
dc.contributor.authorWeinert, Lucyen
dc.contributor.authorLuan, Shiluen
dc.contributor.authorChaudhuri, Roy Ren
dc.contributor.authorAngen, Øysteinen
dc.contributor.authorAragon, Virginiaen
dc.contributor.authorWilliamson, Susanna Men
dc.contributor.authorParkhill, Julianen
dc.contributor.authorLangford, Paul Ren
dc.contributor.authorRycroft, Andrew Nen
dc.contributor.authorWren, Brendan Wen
dc.contributor.authorMaskell, Duncanen
dc.contributor.authorTucker, Alexanderen
dc.date.accessioned2015-09-23T10:39:44Z
dc.date.available2015-09-23T10:39:44Z
dc.date.issued2015-09-30en
dc.identifier.citationJournal of Clinical Microbiology 2015, 53(12):3812-3821. doi:10.1128/JCM.01991-15en
dc.identifier.issn0095-1137
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/251123
dc.description.abstractHaemophilus parasuis causes Glässer’s disease and pneumonia in pigs. Indirect haemagglutination (IHA) is used to serotype this bacterium, distinguishing fifteen serovars with some non-typeable isolates. The capsule loci of the fifteen reference strains were annotated and significant genetic variation was identified between serovars, with the exception of serovars 5 and 12. A capsule locus and in silico serovar was identified for all but two non-typeable isolates in our collection of over 200 isolates. Here we describe the development of a multiplex PCR, based on variation within the capsule loci of the fifteen serovars of H. parasuis, for rapid molecular serotyping. The mPCR distinguished between all previously described serovars except 5 and 12, which were detected by the same pair of primers. The detection limit of the mPCR was 4.29x10⁵ ng/µl bacterial genomic DNA and high specificity was indicated by the absence of reactivity against closely related commensal Pasteurellaceae and other bacterial pathogens of pigs. A subset of 150 isolates from a previously sequenced H. parasuis collection was used to validate the mPCR with 100% accuracy compared to in silico results. In addition, the two in silico non-typeable isolates were typeable using the mPCR. A further 84 isolates were analysed by mPCR and compared to IHA serotyping results with 90% concordance (excluding those non-typeable by IHA). The mPCR was faster, more sensitive, and more specific than IHA enabling the differentiation of fourteen of the fifteen serovars of H. parasuis.
dc.description.sponsorshipThis work was supported by a BPEX PhD studentship and a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1 and BB/G003203/1), the UK Department for Environment, Food and Rural Affairs and Zoetis, awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) consortium. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
dc.languageEnglishen
dc.language.isoenen
dc.publisherAmerican Society for Microbiology
dc.subjectHaemophilus parasuisen
dc.subjectserotypingen
dc.subjectcapsuleen
dc.subjectmolecular serotypingen
dc.titleDevelopment of a multiplex PCR for rapid molecular serotyping of Haemophilus parasuisen
dc.title.alternativeHaemophilus parasuis molecular serotyping assayen
dc.typeArticle
dc.description.versionThis is the author accepted manuscript. The final version is available from American Society for Microbiology via http://dx.doi.org/10.1128/JCM.01991-15en
prism.endingPage3821
prism.publicationDate2015en
prism.publicationNameJournal of Clinical Microbiologyen
prism.startingPage3812
prism.volume53en
dc.rioxxterms.funderBBSRC
dc.rioxxterms.projectidBB/G020744/1
dc.rioxxterms.projectidBB/G019177/1
dc.rioxxterms.projectidBB/G019274/1
dc.rioxxterms.projectidBB/G003203/1
dcterms.dateAccepted2015-09-22en
rioxxterms.versionofrecord10.1128/JCM.01991-15en
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2015-09-30en
dc.contributor.orcidWang, Jinhong [0000-0001-9773-1317]
dc.contributor.orcidHernandez-Garcia, Juan [0000-0001-5932-9327]
dc.contributor.orcidWeinert, Lucy [0000-0002-9279-6012]
dc.contributor.orcidParkhill, Julian [0000-0002-7069-5958]
dc.contributor.orcidMaskell, Duncan [0000-0002-5065-653X]
dc.contributor.orcidTucker, Alexander [0000-0003-0062-0843]
dc.identifier.eissn1098-660X
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBBSRC (BB/G019274/1)
rioxxterms.freetoread.startdate2016-03-30


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