Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , |
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=S1517-83822011000100022 |
Resumo: | In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43), cattle diarrheic faeces (48) and poultry faecal swabs (52) only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30), milk (35), cheese (30), only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each) as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential. |
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Brazilian Journal of Microbiology |
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Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samplesCampylobacter jejuniisolationPCRspikingIn the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43), cattle diarrheic faeces (48) and poultry faecal swabs (52) only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30), milk (35), cheese (30), only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each) as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential.Sociedade Brasileira de Microbiologia2011-03-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000100022Brazilian Journal of Microbiology v.42 n.1 2011reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822011000100022info:eu-repo/semantics/openAccessSingh,HarkanwaldeepRathore,R.SSingh,SatparkashCheema,Pawanjit Singheng2011-01-10T00:00:00Zoai:scielo:S1517-83822011000100022Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2011-01-10T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.fl_str_mv |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
title |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
spellingShingle |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples Singh,Harkanwaldeep Campylobacter jejuni isolation PCR spiking |
title_short |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
title_full |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
title_fullStr |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
title_full_unstemmed |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
title_sort |
Comparative analysis of cultural isolation and PCR based assay for detection of Campylobacter jejuni in food and faecal samples |
author |
Singh,Harkanwaldeep |
author_facet |
Singh,Harkanwaldeep Rathore,R.S Singh,Satparkash Cheema,Pawanjit Singh |
author_role |
author |
author2 |
Rathore,R.S Singh,Satparkash Cheema,Pawanjit Singh |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Singh,Harkanwaldeep Rathore,R.S Singh,Satparkash Cheema,Pawanjit Singh |
dc.subject.por.fl_str_mv |
Campylobacter jejuni isolation PCR spiking |
topic |
Campylobacter jejuni isolation PCR spiking |
description |
In the present study, the efficacy of polymerase chain reaction (PCR) based on mapA gene of C. jejuni was tested for detection of Campylobacter jejuni in naturally infected as well as spiked faecal and food samples of human and animal origin. Simultaneously, all the samples were subjected to the cultural isolation of organism and biochemical characterization. The positive samples resulted in the amplification of a DNA fragment of size ~589 bp in PCR assay whereas the absence of such amplicon in DNA extracted from E. coli, Listeria, Salmonella and Staphylococcus confirmed the specificity of the primers. Of randomly collected 143 faecal samples comprising human diarrheic stools (43), cattle diarrheic faeces (48) and poultry faecal swabs (52) only 4, 3 and 8, respectively, could be detected by isolation whereas 6, 3 and 10, respectively, were found positive by PCR. However, among food samples viz. beef (30), milk (35), cheese (30), only one beef sample was detected both by culture as well as PCR. Additionally, PCR was found to be more sensitive for C. jejuni detection in spiked faecal and food samples (96.1% each) as relative to culture isolation which could detect the organism in 86.7% and 80% samples, respectively. The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high sensitivity, specificity and automation potential. |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011-03-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000100022 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000100022 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1517-83822011000100022 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Brazilian Journal of Microbiology v.42 n.1 2011 reponame:Brazilian Journal of Microbiology instname:Sociedade Brasileira de Microbiologia (SBM) instacron:SBM |
instname_str |
Sociedade Brasileira de Microbiologia (SBM) |
instacron_str |
SBM |
institution |
SBM |
reponame_str |
Brazilian Journal of Microbiology |
collection |
Brazilian Journal of Microbiology |
repository.name.fl_str_mv |
Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM) |
repository.mail.fl_str_mv |
bjm@sbmicrobiologia.org.br||mbmartin@usp.br |
_version_ |
1752122203472658432 |