Abstract
We describe a novel procedure for direct covalent coupling of horseradish peroxidase to rRNA and ribotyping by using enhanced chemiluminescence. Compared with their 32P-end-labeled counterparts, chemiluminescent rRNA probes are stable and easy to synthesize and provide results as good as or superior to those obtained with isotopic labeling. Direct chemiluminescent labeling of Escherichia coli rRNA produces a sensitive, universal probe suitable for clinical laboratory use in the investigation of nosocomial outbreaks.
Full text
PDF


Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Altwegg M., Hickman-Brenner F. W., Farmer J. J., 3rd Ribosomal RNA gene restriction patterns provide increased sensitivity for typing Salmonella typhi strains. J Infect Dis. 1989 Jul;160(1):145–149. doi: 10.1093/infdis/160.1.145. [DOI] [PubMed] [Google Scholar]
- De Buyser M. L., Morvan A., Grimont F., el Solh N. Characterization of Staphylococcus species by ribosomal RNA gene restriction patterns. J Gen Microbiol. 1989 Apr;135(4):989–999. doi: 10.1099/00221287-135-4-989. [DOI] [PubMed] [Google Scholar]
- Forsman M., Sandström G., Jaurin B. Identification of Francisella species and discrimination of type A and type B strains of F. tularensis by 16S rRNA analysis. Appl Environ Microbiol. 1990 Apr;56(4):949–955. doi: 10.1128/aem.56.4.949-955.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goering R. V., Duensing T. D. Rapid field inversion gel electrophoresis in combination with an rRNA gene probe in the epidemiological evaluation of staphylococci. J Clin Microbiol. 1990 Mar;28(3):426–429. doi: 10.1128/jcm.28.3.426-429.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grimont F., Grimont P. A. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Ann Inst Pasteur Microbiol. 1986 Sep-Oct;137B(2):165–175. doi: 10.1016/s0769-2609(86)80105-3. [DOI] [PubMed] [Google Scholar]
- Irino K., Grimont F., Casin I., Grimont P. A. rRNA gene restriction patterns of Haemophilus influenzae biogroup aegyptius strains associated with Brazilian purpuric fever. J Clin Microbiol. 1988 Aug;26(8):1535–1538. doi: 10.1128/jcm.26.8.1535-1538.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LiPuma J. J., Stull T. L., Dasen S. E., Pidcock K. A., Kaye D., Korzeniowski O. M. DNA polymorphisms among Escherichia coli isolated from bacteriuric women. J Infect Dis. 1989 Mar;159(3):526–532. doi: 10.1093/infdis/159.3.526. [DOI] [PubMed] [Google Scholar]
- Owen R. J., Hernandez J., Bolton F. DNA restriction digest and ribosomal RNA gene patterns of Campylobacter jejuni: a comparison with bio-, sero-, and bacteriophage-types of United Kingdom outbreak strains. Epidemiol Infect. 1990 Oct;105(2):265–275. doi: 10.1017/s0950268800047877. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pollard-Knight D., Read C. A., Downes M. J., Howard L. A., Leadbetter M. R., Pheby S. A., McNaughton E., Syms A., Brady M. A. Nonradioactive nucleic acid detection by enhanced chemiluminescence using probes directly labeled with horseradish peroxidase. Anal Biochem. 1990 Feb 15;185(1):84–89. doi: 10.1016/0003-2697(90)90259-c. [DOI] [PubMed] [Google Scholar]
- Rabkin C. S., Jarvis W. R., Anderson R. L., Govan J., Klinger J., LiPuma J., Martone W. J., Monteil H., Richard C., Shigeta S. Pseudomonas cepacia typing systems: collaborative study to assess their potential in epidemiologic investigations. Rev Infect Dis. 1989 Jul-Aug;11(4):600–607. doi: 10.1093/clinids/11.4.600. [DOI] [PubMed] [Google Scholar]
- Rehnstam A. S., Norqvist A., Wolf-Watz H., Hagström A. Identification of Vibrio anguillarum in fish by using partial 16S rRNA sequences and a specific 16S rRNA oligonucleotide probe. Appl Environ Microbiol. 1989 Aug;55(8):1907–1910. doi: 10.1128/aem.55.8.1907-1910.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snipes K. P., Hirsh D. C., Kasten R. W., Hansen L. M., Hird D. W., Carpenter T. E., McCapes R. H. Use of an rRNA probe and restriction endonuclease analysis to fingerprint Pasteurella multocida isolated from turkeys and wildlife. J Clin Microbiol. 1989 Aug;27(8):1847–1853. doi: 10.1128/jcm.27.8.1847-1853.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Stull T. L., LiPuma J. J., Edlind T. D. A broad-spectrum probe for molecular epidemiology of bacteria: ribosomal RNA. J Infect Dis. 1988 Feb;157(2):280–286. doi: 10.1093/infdis/157.2.280. [DOI] [PubMed] [Google Scholar]