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

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.

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Main Authors: Baccaro,Maria Regina, Moreno,Andrea Micke, Shinya,Luciane Tieko, Dotto,Daniela Sabatini
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Microbiologia 2003
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000300008
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spelling oai:scielo:S1517-838220030003000082004-08-04Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigsBaccaro,Maria ReginaMoreno,Andrea MickeShinya,Luciane TiekoDotto,Daniela Sabatini 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.info:eu-repo/semantics/openAccessSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology v.34 n.3 20032003-07-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000300008en10.1590/S1517-83822003000300008
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country Brasil
countrycode BR
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region America del Sur
libraryname SciELO
language English
format Digital
author Baccaro,Maria Regina
Moreno,Andrea Micke
Shinya,Luciane Tieko
Dotto,Daniela Sabatini
spellingShingle Baccaro,Maria Regina
Moreno,Andrea Micke
Shinya,Luciane Tieko
Dotto,Daniela Sabatini
Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs
author_facet Baccaro,Maria Regina
Moreno,Andrea Micke
Shinya,Luciane Tieko
Dotto,Daniela Sabatini
author_sort Baccaro,Maria Regina
title Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs
title_short Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs
title_full Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs
title_fullStr Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs
title_full_unstemmed Identification of bacterial agents of enteric diseases by multiplex PCR in growing-finishing pigs
title_sort identification of bacterial agents of enteric diseases by multiplex pcr in growing-finishing pigs
description 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.
publisher Sociedade Brasileira de Microbiologia
publishDate 2003
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000300008
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