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The capsule loci of the 15 reference strains have been annotated, and significant genetic variation was identified between serovars, with the exception of serovars 5 and 12. A capsule locus and<jats:italic>in silico<\/jats:italic>serovar were identified for all but two nontypeable isolates in our collection of &gt;200 isolates. Here, we describe the development of a multiplex PCR, based on variation within the capsule loci of the 15 serovars of<jats:named-content content-type=\"genus-species\">H. parasuis<\/jats:named-content>, for rapid molecular serotyping. The multiplex PCR (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 4.29 \u00d7 10<jats:sup>5<\/jats:sup>ng\/\u03bcl bacterial genomic DNA, and high specificity was indicated by the absence of reactivity against closely related commensal<jats:named-content content-type=\"genus-species\">Pasteurellaceae<\/jats:named-content>and other bacterial pathogens of pigs. A subset of 150 isolates from a previously sequenced<jats:named-content content-type=\"genus-species\">H. parasuis<\/jats:named-content>collection was used to validate the mPCR with 100% accuracy compared to the<jats:italic>in silico<\/jats:italic>results. In addition, the two<jats:italic>in silico<\/jats:italic>-nontypeable isolates were typeable using the mPCR. A further 84 isolates were analyzed by mPCR and compared to the IHA serotyping results with 90% concordance (excluding those that were nontypeable by IHA). The mPCR was faster, more sensitive, and more specific than IHA, enabling the differentiation of 14 of the 15 serovars of<jats:named-content content-type=\"genus-species\">H. parasuis<\/jats:named-content>.<\/jats:p>","DOI":"10.1128\/jcm.01991-15","type":"journal-article","created":{"date-parts":[[2015,10,1]],"date-time":"2015-10-01T02:29:19Z","timestamp":1443666559000},"page":"3812-3821","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":91,"title":["Development of a Multiplex PCR Assay for Rapid Molecular Serotyping of Haemophilus parasuis"],"prefix":"10.1128","volume":"53","author":[{"given":"Kate J.","family":"Howell","sequence":"first","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah E.","family":"Peters","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinhong","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Hernandez-Garcia","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucy A.","family":"Weinert","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shi-Lu","family":"Luan","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roy R.","family":"Chaudhuri","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield, Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00d8ystein","family":"Angen","sequence":"additional","affiliation":[{"name":"Norwegian Veterinary Institute, Oslo, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virginia","family":"Aragon","sequence":"additional","affiliation":[{"name":"Centre de Recerca en Sanitat Animal (CReSA), UAB-IRTA, Campus de la Universitat Aut\u00f2noma de Barcelona, Bellaterra, and Institut de Recerca i Tecnologia Agroaliment\u00e0ries (IRTA), Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanna M.","family":"Williamson","sequence":"additional","affiliation":[{"name":"Animal and Plant Health Agency (APHA), Suffolk, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7069-5958","authenticated-orcid":false,"given":"Julian","family":"Parkhill","sequence":"additional","affiliation":[{"name":"The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul R.","family":"Langford","sequence":"additional","affiliation":[{"name":"Section of Paediatrics, Department of Medicine, Imperial College London, St. Mary's Campus, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew N.","family":"Rycroft","sequence":"additional","affiliation":[{"name":"The Royal Veterinary College, Hawkshead Campus, Hatfield, Hertfordshire, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brendan W.","family":"Wren","sequence":"additional","affiliation":[{"name":"Faculty of Infectious &amp; Tropical Diseases, London School of Hygiene &amp; Tropical Medicine, London, Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Duncan J.","family":"Maskell","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander W.","family":"Tucker","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2009.10.025"},{"key":"e_1_3_3_3_2","first-page":"681","volume-title":"Diseases of swine","author":"Rapp-Gabrielson VJ","year":"2006","unstructured":"Rapp-Gabrielson VJ, Oliveira SR, Pijoan C. 2006. 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