Rapid identification of Campylobacter species using oligonucleotide probes to 16S ribosomal RNA. 1989

P J Romaniuk, and T J Trust
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.

A comparison of the 16S ribosomal RNA sequences from various Campylobacter species was used to identify unique sequences which distinguish pathogenically significant campylobacters from other members of the genus. Oligonucleotides complementary to these sequences were synthesized, and under stringent conditions hybridization reactions using these probes and total RNA as the target nucleic acid displayed the desired species specificity. A simple, rapid lysis procedure was developed that allowed the specific detection of C. jejuni or C. coli from as few as 10(6) bacterial cells in less than eight hours. This method provides immediate advantages for the unequivocal identification of Campylobacter species in the microbiology laboratory, and demonstrates the potential for the use of these probes in a rapid diagnostic test of clinical samples for Campylobacter infection.

UI MeSH Term Description Entries
D009693 Nucleic Acid Hybridization Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503) Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D002167 Campylobacter A genus of bacteria found in the reproductive organs, intestinal tract, and oral cavity of animals and man. Some species are pathogenic.
D002169 Campylobacter Infections Infections with bacteria of the genus CAMPYLOBACTER. Campylobacter Infection,Campylobacteriosis,Infections, Campylobacter,Campylobacterioses,Infection, Campylobacter
D012335 RNA, Ribosomal The most abundant form of RNA. Together with proteins, it forms the ribosomes, playing a structural role and also a role in ribosomal binding of mRNA and tRNAs. Individual chains are conventionally designated by their sedimentation coefficients. In eukaryotes, four large chains exist, synthesized in the nucleolus and constituting about 50% of the ribosome. (Dorland, 28th ed) Ribosomal RNA,15S RNA,RNA, 15S
D012336 RNA, Ribosomal, 16S Constituent of 30S subunit prokaryotic ribosomes containing 1600 nucleotides and 21 proteins. 16S rRNA is involved in initiation of polypeptide synthesis. 16S Ribosomal RNA,16S rRNA,RNA, 16S Ribosomal,Ribosomal RNA, 16S,rRNA, 16S
D015345 Oligonucleotide Probes Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin. Oligodeoxyribonucleotide Probes,Oligonucleotide Probe,Oligoribonucleotide Probes,Probe, Oligonucleotide,Probes, Oligodeoxyribonucleotide,Probes, Oligonucleotide,Probes, Oligoribonucleotide

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