Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan

Detalhes bibliográficos
Autor(a) principal: Saeed,MA
Data de Publicação: 2022
Outros Autores: Syed,EH, Ghafor,A, Yaqub,T, Javeed,A, Waheed,U
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Poultry Science (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2022000200305
Resumo: ABSTRACT Salmonella Enteritidis (SE) is a dominant serotype among non-typhoidal Salmonella which renders poultry products unsafe for human consumption. Due to frequent reporting of egg associated outbreaks, broiler breeder flocks are understudied although farm environment present supporting conditions for the growth of SE. In this study, two rapid detection techniques for SE were compared in terms of analytical sensitivity and the extent of SE contamination in broiler breeder farm environment was determined. Analytical sensitivity as limit of detection (LOD) was evaluated quantitatively for serotype specific PCR based on amplification of Sdf I gene and a commercially available sandwich ELISA for antigen detection. In triplicate experiments, tenfold serial dilutions of SE were prepared and tested with each technique. Using pure cultures, analytical sensitivity of PCR and ELISA were found to be 18.6 CFU/ml and 2.77×105 CFU/ml respectively. PCR (LOD, log 1.2) was found to be more sensitive and rapid than ELISA (LOD, log 5.4). Environmental swab samples (n = 260) were collected from 22 hen houses representing 8 broiler breeder farms located in and around Lahore and Sheikhupura districts of Punjab province. From each hen house swab samples were collected from litter, nests, feeders, drinkers, fans, pads, ceiling, walls and walkways. Following selective enrichment, pooled swab samples were subjected to PCR. Results showed that 36.3 % (8/22) hen houses were detected positive for SE. These findings suggest improvement in farm biosecurity measures and advocate implementation of integrated Salmonellosis control programs in broiler breeder houses to minimize carcass contamination.
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spelling Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, PakistanSalmonella EnteritidisAnalytical SensitivityOn-farm PrevalenceABSTRACT Salmonella Enteritidis (SE) is a dominant serotype among non-typhoidal Salmonella which renders poultry products unsafe for human consumption. Due to frequent reporting of egg associated outbreaks, broiler breeder flocks are understudied although farm environment present supporting conditions for the growth of SE. In this study, two rapid detection techniques for SE were compared in terms of analytical sensitivity and the extent of SE contamination in broiler breeder farm environment was determined. Analytical sensitivity as limit of detection (LOD) was evaluated quantitatively for serotype specific PCR based on amplification of Sdf I gene and a commercially available sandwich ELISA for antigen detection. In triplicate experiments, tenfold serial dilutions of SE were prepared and tested with each technique. Using pure cultures, analytical sensitivity of PCR and ELISA were found to be 18.6 CFU/ml and 2.77×105 CFU/ml respectively. PCR (LOD, log 1.2) was found to be more sensitive and rapid than ELISA (LOD, log 5.4). Environmental swab samples (n = 260) were collected from 22 hen houses representing 8 broiler breeder farms located in and around Lahore and Sheikhupura districts of Punjab province. From each hen house swab samples were collected from litter, nests, feeders, drinkers, fans, pads, ceiling, walls and walkways. Following selective enrichment, pooled swab samples were subjected to PCR. Results showed that 36.3 % (8/22) hen houses were detected positive for SE. These findings suggest improvement in farm biosecurity measures and advocate implementation of integrated Salmonellosis control programs in broiler breeder houses to minimize carcass contamination.Fundacao de Apoio a Ciência e Tecnologia Avicolas2022-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2022000200305Brazilian Journal of Poultry Science v.24 n.2 2022reponame:Brazilian Journal of Poultry Science (Online)instname:Fundação APINCO de Ciência e Tecnologia Avícolas (FACTA)instacron:FACTA10.1590/1806-9061-2021-1492info:eu-repo/semantics/openAccessSaeed,MASyed,EHGhafor,AYaqub,TJaveed,AWaheed,Ueng2022-04-18T00:00:00Zoai:scielo:S1516-635X2022000200305Revistahttp://www.scielo.br/rbcahttps://old.scielo.br/oai/scielo-oai.php||rvfacta@terra.com.br1806-90611516-635Xopendoar:2022-04-18T00:00Brazilian Journal of Poultry Science (Online) - Fundação APINCO de Ciência e Tecnologia Avícolas (FACTA)false
dc.title.none.fl_str_mv Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
title Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
spellingShingle Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
Saeed,MA
Salmonella Enteritidis
Analytical Sensitivity
On-farm Prevalence
title_short Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
title_full Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
title_fullStr Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
title_full_unstemmed Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
title_sort Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan
author Saeed,MA
author_facet Saeed,MA
Syed,EH
Ghafor,A
Yaqub,T
Javeed,A
Waheed,U
author_role author
author2 Syed,EH
Ghafor,A
Yaqub,T
Javeed,A
Waheed,U
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Saeed,MA
Syed,EH
Ghafor,A
Yaqub,T
Javeed,A
Waheed,U
dc.subject.por.fl_str_mv Salmonella Enteritidis
Analytical Sensitivity
On-farm Prevalence
topic Salmonella Enteritidis
Analytical Sensitivity
On-farm Prevalence
description ABSTRACT Salmonella Enteritidis (SE) is a dominant serotype among non-typhoidal Salmonella which renders poultry products unsafe for human consumption. Due to frequent reporting of egg associated outbreaks, broiler breeder flocks are understudied although farm environment present supporting conditions for the growth of SE. In this study, two rapid detection techniques for SE were compared in terms of analytical sensitivity and the extent of SE contamination in broiler breeder farm environment was determined. Analytical sensitivity as limit of detection (LOD) was evaluated quantitatively for serotype specific PCR based on amplification of Sdf I gene and a commercially available sandwich ELISA for antigen detection. In triplicate experiments, tenfold serial dilutions of SE were prepared and tested with each technique. Using pure cultures, analytical sensitivity of PCR and ELISA were found to be 18.6 CFU/ml and 2.77×105 CFU/ml respectively. PCR (LOD, log 1.2) was found to be more sensitive and rapid than ELISA (LOD, log 5.4). Environmental swab samples (n = 260) were collected from 22 hen houses representing 8 broiler breeder farms located in and around Lahore and Sheikhupura districts of Punjab province. From each hen house swab samples were collected from litter, nests, feeders, drinkers, fans, pads, ceiling, walls and walkways. Following selective enrichment, pooled swab samples were subjected to PCR. Results showed that 36.3 % (8/22) hen houses were detected positive for SE. These findings suggest improvement in farm biosecurity measures and advocate implementation of integrated Salmonellosis control programs in broiler breeder houses to minimize carcass contamination.
publishDate 2022
dc.date.none.fl_str_mv 2022-01-01
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=S1516-635X2022000200305
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2022000200305
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/1806-9061-2021-1492
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 Fundacao de Apoio a Ciência e Tecnologia Avicolas
publisher.none.fl_str_mv Fundacao de Apoio a Ciência e Tecnologia Avicolas
dc.source.none.fl_str_mv Brazilian Journal of Poultry Science v.24 n.2 2022
reponame:Brazilian Journal of Poultry Science (Online)
instname:Fundação APINCO de Ciência e Tecnologia Avícolas (FACTA)
instacron:FACTA
instname_str Fundação APINCO de Ciência e Tecnologia Avícolas (FACTA)
instacron_str FACTA
institution FACTA
reponame_str Brazilian Journal of Poultry Science (Online)
collection Brazilian Journal of Poultry Science (Online)
repository.name.fl_str_mv Brazilian Journal of Poultry Science (Online) - Fundação APINCO de Ciência e Tecnologia Avícolas (FACTA)
repository.mail.fl_str_mv ||rvfacta@terra.com.br
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