Genetic Organization of<i>Pasteurella multocida cap</i>Loci and Development of a Multiplex Capsular PCR Typing System

Townsend, K. M.; Boyce, John D.; Chung, Jing Y.; Frost, A. J.; Adler, Ben

doi:10.1128/jcm.39.3.924-929.2001

Cited by 399 publications

(325 citation statements)

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“…The capsular types were determined by multiplex capsular PCR typing (Townsend et al, 2001). Capsulespecific primers (CAPA, CAPB, CAPD, CAPE and CAPF) were synthesized by Sigma-GenoSys (Cambridge, UK) and the capsular gene fragments were amplified with a Taq DNA polymerase kit (Boehringer Mannheim) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

“…Pooled PCR amplicons of serotype A, B, D, E and F reference strains were used as internal size standards in each gel. Strains that were negative for all five capsular types were confirmed as being P. multocida in separate PCR assays with a P. multocida-specific primer set (Townsend et al, 2001) and classified as untypable.PCR detection of the toxA gene. Detection of the toxA gene was carried out by PCR as described previously (Lichtensteiger et al, 1996) with the exception that different oligonucleotide primers were used.…”

mentioning

confidence: 99%

“…In addition, selected strains representing the various OMP-types were confirmed as P. multocida by sequence analysis of the 16S rRNA gene (unpublished data). The PCR-based capsular typing assay (Townsend et al, 2001) was found to be a rapid and extremely reliable method for determining the capsular types of a large number of P. multocida strains. Of 125 isolates associated with pneumonia, 101 (81 %) were of capsular type A, 20 (16 %) were of capsular type D, two (1 %) were of capsular type F and two (1 %) were untypable.…”

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Characterization and comparison of Pasteurella multocida strains associated with porcine pneumonia and atrophic rhinitis

Davies

MacCorquodale

Baillie

et al. 2003

Journal of Medical Microbiology

104

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One hundred and fifty-eight porcine strains of Pasteurella multocida, recovered primarily from cases of pneumonic pasteurellosis or progressive atrophic rhinitis (PAR) in England and Wales, were characterized by determination of their capsular types, presence or absence of the toxA gene and molecular mass heterogeneity of the heat-modifiable (OmpA) and porin (OmpH) proteins. Eighteen groups (clones) of strains were identified on the basis of specific combinations of capsular type, toxA status and outer-membrane protein (OMP)-type. The data provided evidence that different subpopulations of P. multocida are responsible for pneumonia and PAR in pigs. The majority (88 %) of cases of pneumonia were associated exclusively with non-toxigenic capsular type A strains of OMP-types 1.1, 2.1, 3.1 and 5.1 and capsular type D isolates of OMP-type 6.1. These strains were recovered from widespread geographical locations within England and Wales over a 12-year period and represented mostly single sporadic cases. The association of a small number of P. multocida variants with the majority of cases of porcine pneumonia suggests that these strains are not opportunistic pathogens of low virulence but represent primary pathogens with a relatively high degree of virulence. In contrast, the majority (76 %) of cases of PAR were associated with toxAcontaining capsular type D strains of OMP-type 4.1 and capsular type A and D strains of OMP-type 6.1. Toxigenic capsular type A strains associated with PAR and non-toxigenic capsular type A strains associated with pneumonia represent distinct subpopulations of P. multocida that can be differentiated by their OMP-types. The association of capsular types A and D with strains of the same OMP-types, and the absence and presence of the toxA gene in strains of the same OMP-types, suggest that horizontal transfer of capsular biosynthesis and toxA genes has occurred between strains representing certain subpopulations of P. multocida. INTRODUCTIONPasteurella multocida is the aetiological agent of progressive atrophic rhinitis (PAR) of swine and is of considerable economic importance to the pig-rearing industry throughout the world (Chanter & Rutter, 1989). PAR is characterized by atrophy of the nasal turbinate bones which, in severe cases, can lead to facial distortion. P. multocida strains associated with PAR usually produce a 145-kDa dermonecrotic toxin, which is encoded by the toxA gene (Lax et al., 1990;Buys et al., 1990). This toxin induces osteolysis in the turbinate bones and plays an important role in the pathogenesis of PAR (Kamp & Kimman, 1988). Toxigenic strains associated with PAR are most frequently of capsular type D (Eamens et al., 1988;Foged et al., 1988;Gardner et al., 1994;Lariviere et al., 1992), although toxigenic isolates of capsular type A can also be involved in the disease (Fussing et al., 1999;Sakano et al., 1992). Non-toxigenic P. multocida strains are not usually associated with PAR and confirmation of toxin production is important for the diagnosis and control of the disea...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Characterization and comparison of Pasteurella multocida strains associated with porcine pneumonia and atrophic rhinitis

Davies

MacCorquodale

Baillie

et al. 2003

Journal of Medical Microbiology

104

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“…As amostras de DNA foram eluídas em 100 mL de água ultrapura e armazenadas a -20°C até a reação de polimerase em cadeia (PCR). Foram usadas PCR previamente descritas para amplificação de fragmento do gene da toxina Apx IV de A. pleuropneumoniae (Schaller et al 2001), para o gene 16S rRNA da P. multocida (Townsend et al 2001), para o gene 16S rRNA de H. parasuis (Oliveira et al 2001), para o gene 16S rRNA de M. hyopneumoniae (Yamaguti et al 2008) e para o gene 16S rRNA de M. hyorhinis (Stakenborg et al 2006). Foram consideradas positivas para os agentes as amostras que amplificaram fragmentos dos genes estudados.…”

Section: Methodsunclassified

Pericardite em suínos ao abate no Rio Grande Sul: avaliação de agentes bacterianos e lesões associadas

et al. 2014

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Pesq. Vet. Bras. 34 (7) The objective of the study was to identify the frequency of macroscopic and microscopic lesions and bacterial agents involved with pericarditis in slaughter pigs in the State of Rio Grande do Sul, Brazil. The samples were collected in slaughterhouses with Federal Inspection Service (SIF) between February and October, 2010. Condemnation due to pericarditis in the examined animals was 3.9% (299/7,571). Ninety one cases of pericarditis were examined and by histopathology 89% were chronic and 47% of the corresponding lungs showed chronic pleuritis, but there was no significant association between both lesions. The bacterial agents isolated from the hearts were Streptococcus spp., Pasteurella multocida, Haemophilus parasuis and Streptococcus suis. Bacterial DNA from Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae were the most frequently detected by PCR. There was significant association between isolation of P. multocida and Streptococcus spp. in the hearts and corresponding lungs. The results suggest that lung infection could act as a port of entry to the colonization of the adjacent pericardium. In spite of the fact that M. hyopneumoniae was the agent more frequently identified by PCR in the heart and corresponding lung, there was no significant association of the agent in the organs. This suggests that the infections were independent events. The other agents investigated did not show significant association between isolation or DNA detection in heart and corresponding lungs. Another important finding was the presence of coinfection between bacterial agents in 2% of the hearts and by PCR were identified bacterial DNA of two or more agents in 16.5% of the hearts. These results suggest that coinfections in cases of pericarditis need further investigation.

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“…The suspected colonies were classified as P. multocida by means of PCR using species-specific primers (Townsend et al, 1998). Capsular typing was performed by multiplex capsular PCR with the capsule-specific primers pairs (CAPA, CAPB, CAPD, CAPE, and CAPF) designed by Townsend et al (2001). The capsular PCR conditions have been already described elsewhere (Jaglic et al, 2004).…”

Section: Methodsmentioning

confidence: 99%

Characterisation and comparison of Pasteurella multocida isolated from different species in the Czech Republic: capsular PCR typing, ribotyping and dermonecrotoxin production

Jaglič¹,

Kučerová²,

Nedbalcová³

et al. 2005

Vet. Med. - Czech

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ABSTRACT:The aim of this study was to characterise and compare Pasteurella multocida isolates originating from pigs (n = 43), calves (n = 31), rabbits (n = 27), and to a lesser extent from other hosts (n = 6). A total of 107 P. multocida isolates were obtained from various locations in the Czech Republic. They were analysed by capsular PCR typing and ribotyping, and tested for the production of dermonecrotoxin. Most frequently, serogroup A isolates (n = 74) were found, followed by serogroup D (n = 25) and serogroup F (n = 8) isolates. From a total of fifteen different ribotypes (1-15) generated by restriction endonuclease MspI, four ribotypes (1, 3, 4, and 7) were predominant. The prevalence of predominant ribotypes was different in isolates originating from different hosts. Ribotype 1 was characteristic for rabbit isolates, ribotype 3 was primarily found in pig isolates, and ribotype 7 dominated among calf isolates. Sixteen (mainly porcine) isolates produced dermonecrotoxin but significant correlation among ribotypes and dermonecrotoxin production was not observed.

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Genetic Organization ofPasteurella multocida capLoci and Development of a Multiplex Capsular PCR Typing System

Cited by 399 publications

References 12 publications

Characterization and comparison of Pasteurella multocida strains associated with porcine pneumonia and atrophic rhinitis

Characterization and comparison of Pasteurella multocida strains associated with porcine pneumonia and atrophic rhinitis

Pericardite em suínos ao abate no Rio Grande Sul: avaliação de agentes bacterianos e lesões associadas

Characterisation and comparison of Pasteurella multocida isolated from different species in the Czech Republic: capsular PCR typing, ribotyping and dermonecrotoxin production

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