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Periódicos Brasileiros em Medicina Veterinária e Zootecnia

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Development and application of an ERIC-PCR method for genotyping and differentiating Actinobacillus pleuropneumoniae isolates

Simoni, CintiaLuiza Luiza Luiza LuizaZlotowski, PriscilaDriemeier, DavidCeroni da Silva, Sérgio

Background: The pleuropneumonia caused by Actinobacillus pleuropneumoniae is one the most important swine respiratory diseases. Biochemical and serological tests are widely applied for the diagnosis and characterization of this bacterium. However, in some isolates, conflicting results are found. There are at least 15 serotypes with significant differences in virulence that have been identified until now. Moreover, cross reactions between serotypes are not uncommon. The serotype determination from isolates occurring in outbreaks is an important procedure in prophylaxis and control of the disease. The present work focuses on the application of an ERIC-PCR technique for genotyping and differentiation of A. pleuropneumoniae isolates.Materials, Methods & Results: Fifteen reference strains for the recognized A. pleuropneumoniae serotypes were analyzed in this work, alongside with 27 field isolates that had been previously characterized regarding biochemical, serological and molecular features. Total DNA from each sample was purified and subjected to PCR amplification using ERIC-specific primers (ERIC1R and ERIC2). The resulting amplicons were analyzed by agarose gel electrophoresis and their sizes were estimated from the gel images. Bands with similar sizes were identified and used to construct a binary matrix that took into account the presence or absence of individual bands in

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