Detection of salmonellae in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis

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dc.contributor.authorÜnal, Can Bora
dc.contributor.buuauthorÇarlı, Kamil Tayfun
dc.contributor.buuauthorEyigör, Ayşegül
dc.contributor.buuauthorCaner, Vildan
dc.contributor.departmentUludağ Üniversitesi/Veteriner Fakültesi/Besin Hijyeni ve Teknolojisi Bölümü.tr_TR
dc.contributor.orcidhttp://orcid.org/0000-0003-0980-9335tr_TR
dc.contributor.researcheridE-3867-2010tr_TR
dc.contributor.researcheridAAI-1101-2021tr_TR
dc.contributor.researcheridABI-5029-2020tr_TR
dc.date.accessioned2021-07-05T06:54:06Z
dc.date.available2021-07-05T06:54:06Z
dc.date.issued2001-05
dc.description.abstractThis report describes a rapid detection procedure for salmonellae from chicken feces by the combination of tetrathionate primary enrichment (preenrichment [PE])-bacterial lysis capillary PCR and capillary gel electrophoresis. Pure Salmonella enterica serovar Enteritidis 64K was reisolated and detected by capillary PCR after buffered peptone water and nutrient broth, tetrathionate broth base Hajna (TTBH), and tetrathionate broth (TTB) preenrichments. When the same culture was mixed with intestinal homogenate, bacteriological reisolation and capillary PCR detection was achieved only by TTBK and TTB preenrichments. Capillary gel electrophoresis revealed that a Salmonella genus-specific 281-bp PCR product aas detected when Salmonella strains but not non-Salmonella strains were tested. The detection limit of capillary PCR with whole-cell DNA extracted from pure Salmonella enterica serovars Enteritidis 64K, Typhimurium LT2-CIP60-62, and Gallinarum 64K was 3, 3, and 9 CPU ml(-1), respectively. The detection limit of capillary PCR from whole cell DNA extracted from intestinal homogenate artificially contaminated with the same three strains was 3, 3, and 7 CFU ml(-1), respectively We compared the results of the capillary PCR and bacteriological examination from the natural samples. Thirty-five of 9 naturally contaminated samples produced a specific PCR product. In 9 of the 35 PCR-positive samples, Salmonella could not be detected bacteriologically either by PE or a primary and delayed secondary enrichment (DSE) combination. In the 18 PCR-negative samples, 4 samples were found to harbor Salmonella by both PE and DSE and 14 samples were positive after DSE. Fifty-three additional intestinal homogenate samples, which were negative by their PE and DSE in bacteriological examination, were found to be also negative by their PCRs. The total time required to detect Salmonella with the capillary PCR method we used was approximately 20 h, If samples are from clinically diseased birds, the total time for PCR and detection is reduced to 2 h since the 18-h PE is not required. These results indicate that TTB enrichment, bacterial lysis, and genus-specific capillary PCR combined,vith capillary gel electrophoresis constitute a sensitive and selective procedure which has the potential to rapidly identify Salmonella-infected flocks.en_US
dc.identifier.citationÇarlı, K. T. vd. (2001). " Detection of salmonellae in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis". Journal of Clinical Microbiology, 39(5), 1871-1876.tr_TR
dc.identifier.endpage1876tr_TR
dc.identifier.issn0095-1137
dc.identifier.issue5tr_TR
dc.identifier.pubmed11326006tr_TR
dc.identifier.scopus2-s2.0-0035001706tr_TR
dc.identifier.startpage1871tr_TR
dc.identifier.urihttps://doi.org/10.1128/JCM.39.5.1871-1876.2001
dc.identifier.urihttps://journals.asm.org/doi/10.1128/JCM.39.5.1871-1876.2001
dc.identifier.urihttp://hdl.handle.net/11452/21037
dc.identifier.volume39tr_TR
dc.identifier.wos000168527900033tr_TR
dc.indexed.pubmedPubmeden_US
dc.indexed.scopusScopusen_US
dc.indexed.wosSCIEen_US
dc.language.isoenen_US
dc.publisherAmer Soc Microbiologyen_US
dc.relation.collaborationYurtiçitr_TR
dc.relation.journalJournal of Clinical Microbiologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergitr_TR
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectSerogroup-Den_US
dc.subjectPolymerase-chain-reactionen_US
dc.subjectCycle dna amplificationen_US
dc.subjectImmunomagnetic separationen_US
dc.subjectEnvironmental-samplesen_US
dc.subjectTyphimuriumen_US
dc.subjectAssayen_US
dc.subjectEnteritidisen_US
dc.subjectProductsen_US
dc.subjectMeaten_US
dc.subject.wosMicrobiologyen_US
dc.titleDetection of salmonellae in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresisen_US
dc.typeArticle
dc.wos.quartileQ1en_US

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