Identification of Campylobacter jejuni, Campylobacter coli and campylobacter lari by the nucleic acid amplification system NASBAR

J Appl Bacteriol. 1994 Dec;77(6):694-701. doi: 10.1111/j.1365-2672.1994.tb02821.x.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Campylobacter / classification*
  • Campylobacter / genetics
  • Campylobacter / isolation & purification
  • Campylobacter coli / classification
  • Campylobacter coli / genetics
  • Campylobacter coli / isolation & purification
  • Campylobacter jejuni / classification
  • Campylobacter jejuni / genetics
  • Campylobacter jejuni / isolation & purification
  • DNA Primers
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques*
  • RNA, Ribosomal, 16S / genetics
  • RNA, Ribosomal, 16S / isolation & purification*
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • RNA, Ribosomal, 16S