Development of a real-time NASBA assay for the detection of Campylobacter jejuni cells.
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.
Cools
Ils
I
Laboratory of Food Microbiology and Food Preservation, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, B-9000 Gent, Belgium.
Uyttendaele
Mieke
M
D'Haese
Eva
E
Nelis
Hans J
HJ
Debevere
Johan
J
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
D012329:RNA, Bacterial; D012333:RNA, Messenger; D020558:GTP Phosphohydrolases; D010444:Peptide Elongation Factor Tu