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dc.contributor.authorAbu-Halaweh, Marwan
dc.contributor.authorBates, J
dc.contributor.authorPatel, Bharat KC
dc.date.accessioned2017-05-03T11:40:31Z
dc.date.available2017-05-03T11:40:31Z
dc.date.issued2005
dc.date.modified2009-09-16T07:43:10Z
dc.identifier.issn0923-2508
dc.identifier.doi10.1016/j.resmic.2004.08.008
dc.identifier.urihttp://hdl.handle.net/10072/4901
dc.description.abstractA two-tube real-time assay, developed in a LightCyclerTM, was used to detect, identify and differentiate Campylobacter jejuni and Campylobacter coli from all other pathogenic members of the family Campylobacteriaceae. In the first assay, continuous monitoring of the fluorescence resonance energy transfer (FRET) signal acquired from the hybridisation of two adjacent fluoroprobes, a specific FITC probe 5'-GTGCTAGCTTGCTAGAACTTAGAGA-FITC-3') and a universal downstream probe Cy5 (5'-Cy5-AGGTGITGCATGGITGTCGTTGTCG-PO4-3'), to the 681-base pair 16S rRNA gene amplicon target (Escherichia coli position 1024-1048 and 1050-1075, respectively) produced by the primer pair, F2 (ATCTAATGGCTTAACCATTAAAC, E. coli position 783) and Cam-Rev (AATACTAAACTAGTTACCGTC, E. coli position 1464), detected C. coli, C. lari and C. jejuni. As expected, a Tm of 65 àwas derived from the temperature-dependent probe DNA strand disassociation. In the second assay, an increase in fluorescence due to binding of the intercalating dye SYBR Green I to the DNA amplicons of the hippuricase gene (hipO) (produced by the primer pair hip2214F and hip2474R) was observed for C. jejuni but not for C. coli which lacks the hipO gene. A Tm of 85ᰮ5 and 56 àdetermined from temperature-dependent dye-DNA disassociation identified C. jejuni and the non-specific PCR products, respectively, in line with our expectation. The two-tube assay was subsequently used to identify and differentiate the 169 Campylobacteriaceae isolates of animal, human, plant and bird origin held in our culture collection into C. coli (74 isolates), C. jejuni (86 isolates) and non-C. coli-C. jejuni (9 isolates). In addition, the method successfully detected C. jejuni, C. coli and C. lari from 24-h enrichment cultures initiated from 30 commercial chicken samples.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherElsevier
dc.publisher.placeFrance
dc.publisher.urihttp://www.elsevier.com/wps/find/journaldescription.cws_home/522493/description#description
dc.relation.ispartofstudentpublicationY
dc.relation.ispartofpagefrom107
dc.relation.ispartofpageto114
dc.relation.ispartofjournalResearch in Microbiology
dc.relation.ispartofvolume156
dc.rights.retentionY
dc.subject.fieldofresearchMicrobiology
dc.subject.fieldofresearchMedical microbiology
dc.subject.fieldofresearchcode3107
dc.subject.fieldofresearchcode3207
dc.titleRapid detection and identification of pathogenic Campylobacter jejuni and Campylobacter coli by real-time PCR
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.facultyGriffith Sciences, School of Natural Sciences
gro.date.issued2005
gro.hasfulltextNo Full Text
gro.griffith.authorPatel, Bharat K.
gro.griffith.authorAbu-Halaweh, Marwan


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