Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis

Bovine genital campylobacteriosis is a common venereal disease of cattle; the prevalence of this disease can be underestimated mostly because of the nature of the etiological agent, the microaerobic Campylobacter fetus subspecies venerealis. The purpose of the current study was to evaluate the utilization of polymerase chain reaction (PCR) in the diagnosis of genital campylobacteriosis in samples obtained from bull prepuce aspirate, cow cervical mucus, and abomasum contents of aborted fetuses, collected into enrichment medium. Five different DNA extraction protocols were tested: thermal extraction, lysis with proteinase K, lysis with guanidine isothiocyanate, lysis with DNAzol, and lysis with hexadecyltrimethylammonium bromide (CTAB). The specificity, sensitivity, and technical application of the PCR assay were also evaluated with clinical samples and compared to bacterial isolation by standard culture. DNA extraction by the CTAB protocol provided better results in PCR, and it was able to detect 63 colony-forming units per ml of C. fetus. Out of 277 clinical samples tested, 68 (24%) were positive for Campylobacter fetus using PCR, while only 8 (2.8%) of the samples were positive by bacterial isolation in solid medium, proving the superiority of the PCR technique when compared to the standard isolation method, and providing evidence for its usefulness as a better screening test in cattle for the diagnosis of bovine genital campylobacteriosis.

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Main Authors: Groff,Ana C.M., Kirinus,Jackeline K., Silva,Mariana Sá e, Machado,Gustavo, Costa,Mateus M., Vargas,Agueda P.C.
Format: Digital revista
Language:English
Published: Colégio Brasileiro de Patologia Animal - CBPA 2010
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2010001200005
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spelling oai:scielo:S0100-736X20100012000052011-01-31Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosisGroff,Ana C.M.Kirinus,Jackeline K.Silva,Mariana Sá eMachado,GustavoCosta,Mateus M.Vargas,Agueda P.C. Bovine genital campylobacteriosis Campylobacter fetus diagnosis abortion Bovine genital campylobacteriosis is a common venereal disease of cattle; the prevalence of this disease can be underestimated mostly because of the nature of the etiological agent, the microaerobic Campylobacter fetus subspecies venerealis. The purpose of the current study was to evaluate the utilization of polymerase chain reaction (PCR) in the diagnosis of genital campylobacteriosis in samples obtained from bull prepuce aspirate, cow cervical mucus, and abomasum contents of aborted fetuses, collected into enrichment medium. Five different DNA extraction protocols were tested: thermal extraction, lysis with proteinase K, lysis with guanidine isothiocyanate, lysis with DNAzol, and lysis with hexadecyltrimethylammonium bromide (CTAB). The specificity, sensitivity, and technical application of the PCR assay were also evaluated with clinical samples and compared to bacterial isolation by standard culture. DNA extraction by the CTAB protocol provided better results in PCR, and it was able to detect 63 colony-forming units per ml of C. fetus. Out of 277 clinical samples tested, 68 (24%) were positive for Campylobacter fetus using PCR, while only 8 (2.8%) of the samples were positive by bacterial isolation in solid medium, proving the superiority of the PCR technique when compared to the standard isolation method, and providing evidence for its usefulness as a better screening test in cattle for the diagnosis of bovine genital campylobacteriosis.info:eu-repo/semantics/openAccessColégio Brasileiro de Patologia Animal - CBPAPesquisa Veterinária Brasileira v.30 n.12 20102010-12-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2010001200005en10.1590/S0100-736X2010001200005
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language English
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author Groff,Ana C.M.
Kirinus,Jackeline K.
Silva,Mariana Sá e
Machado,Gustavo
Costa,Mateus M.
Vargas,Agueda P.C.
spellingShingle Groff,Ana C.M.
Kirinus,Jackeline K.
Silva,Mariana Sá e
Machado,Gustavo
Costa,Mateus M.
Vargas,Agueda P.C.
Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
author_facet Groff,Ana C.M.
Kirinus,Jackeline K.
Silva,Mariana Sá e
Machado,Gustavo
Costa,Mateus M.
Vargas,Agueda P.C.
author_sort Groff,Ana C.M.
title Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
title_short Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
title_full Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
title_fullStr Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
title_full_unstemmed Polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
title_sort polymerase chain reaction for the diagnosis of bovine genital campylobacteriosis
description Bovine genital campylobacteriosis is a common venereal disease of cattle; the prevalence of this disease can be underestimated mostly because of the nature of the etiological agent, the microaerobic Campylobacter fetus subspecies venerealis. The purpose of the current study was to evaluate the utilization of polymerase chain reaction (PCR) in the diagnosis of genital campylobacteriosis in samples obtained from bull prepuce aspirate, cow cervical mucus, and abomasum contents of aborted fetuses, collected into enrichment medium. Five different DNA extraction protocols were tested: thermal extraction, lysis with proteinase K, lysis with guanidine isothiocyanate, lysis with DNAzol, and lysis with hexadecyltrimethylammonium bromide (CTAB). The specificity, sensitivity, and technical application of the PCR assay were also evaluated with clinical samples and compared to bacterial isolation by standard culture. DNA extraction by the CTAB protocol provided better results in PCR, and it was able to detect 63 colony-forming units per ml of C. fetus. Out of 277 clinical samples tested, 68 (24%) were positive for Campylobacter fetus using PCR, while only 8 (2.8%) of the samples were positive by bacterial isolation in solid medium, proving the superiority of the PCR technique when compared to the standard isolation method, and providing evidence for its usefulness as a better screening test in cattle for the diagnosis of bovine genital campylobacteriosis.
publisher Colégio Brasileiro de Patologia Animal - CBPA
publishDate 2010
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2010001200005
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