Comparison of Genotypic and Phenotypic Characterization Methods forPasteurella MultocidaIsolates from Fatal Cases of Bovine Respiratory Disease

Abstract

Abstract. Bovine respiratory disease (BRD) is the most costly disease of beef cattle in North America. Because Pasteurella multocida is a commensal of the upper respiratory tract, it is generally considered an opportunistic pathogen. However, studies in swine indicated that there may be a limited number of strains associated with disease, suggesting that some are more virulent than others. Although this may also be true of isolates from cattle, appropriate typing methods must be established before this possibility can be investigated. The purpose of this study was to compare effectiveness of polymerase chain reaction (PCR) fingerprinting to more traditional approaches for typing bovine P. multocida isolates. Isolates were obtained from 41 cases of fatal BRD and subjected to random amplified polymorphic DNA PCR (RAPD-PCR), whole cell protein (WCP) profiles, outer membrane protein (OMP) profiles, and serotyping. The discrimination index was calculated for each typing method and combinations of each using Simpson's index of diversity. Correlation coefficients were calculated to assess concordance between classification results achieved through genotypic (RAPD-PCR) and phenotypic (WCP, OMP, and serotyping) approaches. All characterization methods were capable of discriminating between isolates. However, there was poor concordance between techniques. There were also few significant associations between typing results and epidemiologic data. Random amplified polymorphic DNA PCR was validated as being a repeatable and reliable means of discriminating between P. multocida isolates obtained from cattle. Isolates obtained from fatal cases of BRD in calves in a commercial feedlot demonstrated significant diversity, justifying additional investigation into whether P. multocida is a strictly opportunistic pathogen in cattle.

Phenotypic Characterization and Random Amplified Polymorphic DNA (RAPD) analysis of <i>Pasteurella multocida</i> isolated from Korean Pigs

“…The primer used for RAPD-PCR was MM13 (5′-GAGGGTG-GNGGNTCT-3′), as previously described [15]. The PCR conditions consisted of 30 cycles of 30 sec at 95°C, 30 sec at 42°C, and 30 sec at 72°C.…”

“…These characteristics have also been analyzed by ribotyping, single-primer PCR, and Random Amplified Polymorphic DNA (RAPD) PCR [5,12,15]. The merit of the RAPD-PCR assay is that it is simple and requires less time to process than other methods of restriction endonuclease analysis [12,15].…”

“…These characteristics have also been analyzed by ribotyping, single-primer PCR, and Random Amplified Polymorphic DNA (RAPD) PCR [5,12,15]. The merit of the RAPD-PCR assay is that it is simple and requires less time to process than other methods of restriction endonuclease analysis [12,15]. RAPD-PCR analysis for characterization of P. multocida has yielded reproducible and reliable results in comparison with other analytical methods such as whole cell protein, outer membrane protein, serotyping, and restriction endonuclease analyses [15].…”

“…The merit of the RAPD-PCR assay is that it is simple and requires less time to process than other methods of restriction endonuclease analysis [12,15]. RAPD-PCR analysis for characterization of P. multocida has yielded reproducible and reliable results in comparison with other analytical methods such as whole cell protein, outer membrane protein, serotyping, and restriction endonuclease analyses [15]. An ornithological study showed that 66 strains of P. multocida isolated from birds could be easily and quickly categorized into 18 groups through RAPD-PCR [12].…”

Análisis de Diversidad Genética de Cepas de Pasteurella multocida Aisladas de Alpacas con Signos de Neumonía

“…Sin embargo, esta diversidad fue difícil de interpretar porque todas las cepas fueron aisladas de un mismo brote (Taylor et al, 2010).…”