title: Identification of Campylobacter jejuni, Campylobacter coli and campylobacter lari by the nucleic acid amplification system NASBAR.
abstract: NASBAR, an isothermal amplification technique for nucleic acids, was evaluated for the specific identification of Campylobacter jejuni, Camp. coli and Camp. lari. A set of primers and a probe were chosen from the 16S rRNA sequence of Campylobacter. The probe was hybridized in solution with the amplified nucleic acids of 12 Campylobacter species and nine other Gram-negative bacteria. The probe was shown to hybridize specifically to the amplified single-stranded RNA of Camp. jejuni, Camp. coli and Camp. lari in an enzyme-linked gel assay (ELGA). In a Camp. jejuni model system the combination of NASBAR and ELGA was able to detect ca 1000 rRNA molecules. The presence of an excess of Gram-negative bacteria did not influence the sensitivity of detection. A number of 6 cfu of Camp. jejuni, present in a total count of 4 x 10(6) cfu of Gram-negative bacteria, resulted in a positive hybridization signal.
authors: Department of Food Technology and Nutrition, Faculty of Agricultural, UG, Ghent, Belgium.
authors: M
authors: M
authors: Uyttendaele
authors: R
authors: R
authors: Schukkink
authors: B
authors: B
authors: van Gemen
authors: J
authors: J
authors: Debevere
mesh_terms: D001483:Base Sequence; D002167:Campylobacter / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D017000:Campylobacter coli / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D012336:RNA, Ribosomal, 16S