Characterization of the ptfA Gene of Avian Pasteurella Multocida Strains by Allele-Specific Polymerase Chain Reaction

Abstract

Abstract. Pasteurella multocida is the causative agent of fowl cholera in domesticated and wild birds. The disease outcome is affected by various host-and pathogen-specific determinants. Several putative virulence factors have been proposed to play a key role in this interaction, including the ptfA gene, the products of which assemble to form type 4 fimbriae on the bacterial surface. One way to understand more precisely how ptfA contributes to pathogenesis is to gather molecular features of this gene in circulating avian P. multocida strains. Therefore, molecular characterization of the ptfA gene of P. multocida strains isolated from domestic poultry was performed using the combination of nucleotide sequence analysis and a newly developed allelespecific polymerase chain reaction assay. Two major ptfA alleles were identified among 31 strains, representing various serogroups and somatic serotypes. It was noteworthy that allele specificity and case severity of a subset of strains correlated with the available gross pathology data. Therefore, the acquisition of comprehensive clinical and epidemiological data together with molecular characteristics of individual strains will help to design and implement adequate preventive and intervention strategies.

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

“…The Type IV fimbriae are formed by repeated subunits and present a highly conserved N-terminal nucleotide sequence (Hatfaludi et al 2010). In a recent study, Sellyei et al (2010) identified only two major alleles of ptfA among 31 strains of different sero- Doughty et al (2000), this gene is a candidate for heterologous vaccine development (Ewers et al 2006). The hemagglutinin PfhA was identified in 15/25 strains (60%).…”

“…It is believed that among the adhesins identified in samples of P. multocida, including pfhA, tad, ptfA, fimA, and hsf-1,2, only the latter three are present in all pathogenic strains (Sellyei et al 2010). The ptfA gene, which encodes a subunit of type IV fimbriae, was found in 23/25 samples (92%), a result similar to that observed in other studies (Ewers et al 2006, Bethe et al 2009, Tang et al 2009).…”

Sequencing-independent method for the differentiation of the main phylogenetic lineages ofPasteurella multocida

“…Genomic DNA was extracted using a commercial kit c as described previously. 12 The 16S rDNA (1235 nucleotide in length) was amplified by PCR carried out in a 50-μl volume that was comprised of 5 μl of DNA template, 12.5 pmol of each modified universal primer d (forward: 5′-AGAGTTTGATCATGGCTCAG-3′, position 9-29; reverse: 5′-CCATTGTAGGACGTGTGAAGCCC-3′, position 1222-1244) in 1× PCR buffer, e 200 nM deoxyribonucleotide triphosphate mix, e and 1.25 U commercial DNA polymerase.…”

First detection of Pasteurella multocida type B:2 in Hungary associated with systemic pasteurellosis in backyard pigs

“…Genes were detected by PCR as previously described (Tang et al, 2009;Sellyei et al, 2010). All reactions were performed using a C1000 Thermal Cycler (BioRad Laboratories, Inc., Berkeley, CA, USA).…”