Problems with serological cross-reactivity have led to development of a number of PCRs (individual and multiplex) for molecular typing of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. Most of these assays were developed for detection of specific amplicons within capsule biosynthetic genes before the availability of complete sequences for the different serovars. Here we describe comparative analysis of the complete capsular loci for all 18 serovars of A. pleuropneumoniae, and development of two multiplex PCRs for comprehensive capsule typing of this important pig pathogen.

• Klíčová slova
• A. pleuropneumoniae, Capsule typing, Diagnostic, Serovars, mPCR,
• MeSH
• Actinobacillus pleuropneumoniae klasifikace   genetika   patogenita   MeSH
• bakteriální polysacharidy genetika   MeSH
• bakteriální pouzdra chemie   klasifikace   genetika   MeSH
• infekce bakteriemi rodu Actinobacillus diagnóza   mikrobiologie   MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• nemoci prasat diagnóza   mikrobiologie   MeSH
• prasata MeSH
• sekvenční analýza * MeSH
• séroskupina MeSH
• sérotypizace MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• bakteriální polysacharidy MeSH

The aim of this study was to investigate isolates of Actinobacillus pleuropneumoniae previously designated serologically either as non-typable (NT) or as 'K2:07', which did not produce serovar-specific amplicons in PCR assays. We used whole genome sequencing to identify the capsule (CPS) loci of six previously designated biovar 1 NT and two biovar 1 'K2:O7' isolates of A. pleuropneumoniae from Denmark, as well as a recent biovar 2 NT isolate from Canada. All of the NT isolates have the same six-gene type I CPS locus, sharing common cpsABC genes with serovars 2, 3, 6, 7, 8, 9, 11 and 13. The two 'K2:O7' isolates contain a unique three-gene type II CPS locus, having a cpsA gene similar to that of serovars 1, 4, 12, 14 and 15. The previously NT isolates share the same O-antigen genes, found between erpA and rpsU, as serovars 3, 6, 8, and 15. Whereas the 'K2:O7' isolates, have the same O-antigen genes as serovar 7, which likely contributed to their previous mis-identification. All of the NT and 'K2:O7' isolates have only the genes required for production of ApxII (apxIICA structural genes, and apxIBD export genes). Rabbit polyclonal antisera raised against representative isolates with these new CPS loci demonstrated distinct reactivity compared to the 16 known serovars. The serological and genomic results indicate that the isolates constitute new serovars 17 (previously NT) and 18 (previously 'K2:O7'). Primers designed for amplification of specific serovar 17 and 18 sequences for molecular diagnostics will facilitate epidemiological tracking of these two new serovars of A. pleuropneumoniae.

• Klíčová slova
• Actinobacillus pleuropneumoniae, Capsule genes, Diagnostics, PCR, Serovar 17, Serovar 18,
• MeSH
• Actinobacillus pleuropneumoniae klasifikace   genetika   imunologie   izolace a purifikace   MeSH
• bakteriální pouzdra genetika   MeSH
• DNA bakterií genetika   MeSH
• DNA primery genetika   MeSH
• genotyp * MeSH
• infekce bakteriemi rodu Actinobacillus epidemiologie   veterinární   MeSH
• nemoci prasat epidemiologie   mikrobiologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• prasata MeSH
• sekvenování celého genomu MeSH
• séroskupina * MeSH
• sérotypizace MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Dánsko epidemiologie   MeSH
• Kanada epidemiologie   MeSH
• Názvy látek
• DNA bakterií MeSH
• DNA primery MeSH