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Multiplex real-time PCR for detection of Campylobacter, Salmonella, and Shigella

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dc.contributor.author Barletta, F.
dc.contributor.author Mercado, E. H.
dc.contributor.author Lluque, A.
dc.contributor.author Ruiz, J.
dc.contributor.author Cleary, T. G.
dc.contributor.author Ochoa Woodell, Theresa Jean
dc.date.accessioned 2022-01-04T20:31:42Z
dc.date.available 2022-01-04T20:31:42Z
dc.date.issued 2013
dc.identifier.uri https://hdl.handle.net/20.500.12866/10548
dc.description.abstract Infectious diarrhea can be classified based on its clinical presentation as noninflammatory or inflammatory disease. In developing countries, among inflammatory diarrhea cases, Shigella is the most common cause, followed by Campylobacter and Salmonella. Because the time frame in which treatment choices must be made is short and conventional stool cultures lack good sensitivity, there is a need for a rapid, sensitive, and inexpensive detection technique. The purpose of our study was to develop a multiplex real-time PCR procedure to simultaneously identify Campylobacter spp., Salmonella spp., and Shigella spp. Primers were designed to amplify the invA, ipaH, and 16S rRNA genes simultaneously in a single reaction to detect Salmonella, Shigella, and Campylobacter, respectively. Using this approach, we correctly identified 102 of 103 strains of the targeted enteropathogens and 34 of 34 other pathogens. The melting temperatures were 82.96 ± 0.05°C for invA, 85.56 ± 0.28°C for ipaH, and 89.21 ± 0.24°C for 16S rRNA. The limit of accurate quantification for the assay in stool samples was 104 CFU g−1; however, the limit of detection was 103 CFU g−1. This assay is a simple, rapid, inexpensive, and reliable system for the practical detection of these three enteropathogens in clinical specimens. en_US
dc.language.iso eng
dc.publisher American Society for Microbiology
dc.relation.ispartofseries Journal of Clinical Microbiology
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Sensitivity and Specificity en_US
dc.subject Genes en_US
dc.subject Transition Temperature en_US
dc.subject Bacteriological Techniques en_US
dc.subject Campylobacter Infections en_US
dc.subject Campylobacter en_US
dc.subject DNA en_US
dc.subject Dysentery en_US
dc.subject Molecular Diagnostic Techniques en_US
dc.subject Multiplex Polymerase Chain Reaction en_US
dc.subject Real-Time Polymerase Chain Reaction en_US
dc.subject Salmonella Infections en_US
dc.subject Shigella en_US
dc.title Multiplex real-time PCR for detection of Campylobacter, Salmonella, and Shigella en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1128/JCM.01397-13
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#1.06.01
dc.relation.issn 1098-660X


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