Identification of Campylobacter jejuni, Campylobacter coli and campylobacter lari by the nucleic acid amplification system NASBAR.
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.
Uyttendaele
M
M
Department of Food Technology and Nutrition, Faculty of Agricultural, UG, Ghent, Belgium.
Schukkink
R
R
van Gemen
B
B
Debevere
J
J
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
D017931:DNA Primers; D012336:RNA, Ribosomal, 16S