title: Development of a real-time NASBA assay for the detection of Campylobacter jejuni cells.
abstract: The objectives of this study were the development of a real-time NASBA assay for the detection of Campylobacter jejuni mRNA and the evaluation of its potential to determine the viability of the detected C. jejuni cells. A set of specific primers and probes was chosen to amplify the mRNA of the tuf-gene and the GTPase-gene. Only the tuf-assay was able to detect as low as 10(2) cells per NASBA reaction and was specific for Campylobacter. However, as the assay was able to detect dead cells, it cannot be used to demonstrate the viability of C. jejuni cells. The tuf-gene mRNA is no good viability indicator due to its stability.
authors.affiliation: Laboratory of Food Microbiology and Food Preservation, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, B-9000 Gent, Belgium.
authors.forename: Ils
authors.initials: I
authors.lastname: Cools
authors.forename: Mieke
authors.initials: M
authors.lastname: Uyttendaele
authors.forename: Eva
authors.initials: E
authors.lastname: D'Haese
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Nelis
authors.forename: Johan
authors.initials: J
authors.lastname: Debevere
mesh_terms: D001483:Base Sequence; D016123:Campylobacter jejuni / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000235:genetics; D050296:Microbial Viability; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010444:Peptide Elongation Factor Tu / Q000737:chemistry / Q000235:genetics; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D021161:Self-Sustained Sequence Replication / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity
chemical_list: D012329:RNA, Bacterial; D012333:RNA, Messenger; D020558:GTP Phosphohydrolases; D010444:Peptide Elongation Factor Tu