Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs

• Autor(a) principal: Baccaro,Maria Regina
• Data de Publicação: 2003
• Outros Autores: Moreno,Andrea Micke, Shinya,Luciane Tieko, Dotto,Daniela Sabatini
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Brazilian Journal of Microbiology
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000300008
• topic: enteritis; PCR; pig; bacteria; diagnosis

In Brazil, the most common bacterial enteric diseases affecting growing and finishing pigs are porcine proliferative enteritis, porcine intestinal spirochetosis, swine dysentery, and salmonellosis. The diagnosis of these diseases by routine culture techniques is expensive, difficult, time-consuming, and even impossible, in cases of porcine proliferative enteritis. The detection of pathogens by polymerase chain reaction is a highly sensitive and specific method that can be an useful tool in veterinary diagnosis. Two multiplex PCR (M-PCR) assays were tested for simultaneous detection and identification of bacterial agents associated with porcine proliferative enteritis, porcine intestinal spirochetosis, swine dysentery, and salmonellosis in diarrheic fecal samples. The DNA obtained from pure cultures of each bacterial agent or mixed in different combinations and concentrations was amplified by using Lawsonia intracellularis and Salmonella, or Brachyspira pilosicoli and Brachyspira hyodysenteriae specific M-PCR assays. After electrophoresis in agarose gel and staining, the amplification products indicated the presence of individual or simultaneous amplification of L. intracellularis and Salmonella or B. pilosicoli and B. hyodysenteriae specific DNA sequences. After standardization, the M-PCR tests were used to test 541 swine diarrheic fecal samples obtained from different regions in Brazil. The most frequently detected pathogen was Lawsonia intracellularis (13%), followed by Salmonella (4.8%), B. hyodysenteriae (1.4%), B. pilosicoli (1%) and their various associations. Results from this study suggest that the two M-PCR assays can be used for specific detection and identification of four important enteric bacterial pathogens alone or in combination.