Abstract
A novel method for molecular typing of organisms, amplified fragment length polymorphism analysis, was tested for its suitability in epidemiological studies in medical microbiology. Amplified fragment length polymorphism analysis, originally developed for typing crop plants, consists of a simple restriction-ligation reaction and a subsequent PCR amplification. In a single-step reaction, the genomic DNA is digested and the restriction fragments are ligated to specially constructed adapters. PCR amplification of such tagged restriction fragments with primers complementary to the adapters allows the detection of restriction fragment length polymorphisms upon resolution on agarose gels. The method is fast, efficient, and reproducible for typing strains of Legionella pneumophila isolated from both humans and the environment. The accuracy of the method was tested by comparison with standard restriction fragment length polymorphism typing performed with both a ribosomal and a genomic probe.
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Selected References
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- Bingen E. H., Denamur E., Elion J. Use of ribotyping in epidemiological surveillance of nosocomial outbreaks. Clin Microbiol Rev. 1994 Jul;7(3):311–327. doi: 10.1128/cmr.7.3.311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brosius J., Ullrich A., Raker M. A., Gray A., Dull T. J., Gutell R. R., Noller H. F. Construction and fine mapping of recombinant plasmids containing the rrnB ribosomal RNA operon of E. coli. Plasmid. 1981 Jul;6(1):112–118. doi: 10.1016/0147-619x(81)90058-5. [DOI] [PubMed] [Google Scholar]
- Edelstein P. H. Improved semiselective medium for isolation of Legionella pneumophila from contaminated clinical and environmental specimens. J Clin Microbiol. 1981 Sep;14(3):298–303. doi: 10.1128/jcm.14.3.298-303.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ferdows M. S., Barbour A. G. Megabase-sized linear DNA in the bacterium Borrelia burgdorferi, the Lyme disease agent. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5969–5973. doi: 10.1073/pnas.86.15.5969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gaia V., Poloni C., Peduzzi R. Epidemiological typing of Legionella pneumophila with ribotyping. Report of two clinical cases. Eur J Epidemiol. 1994 Jun;10(3):303–306. doi: 10.1007/BF01719354. [DOI] [PubMed] [Google Scholar]
- Grimont F., Lefèvre M., Ageron E., Grimont P. A. rRNA gene restriction patterns of Legionella species: a molecular identification system. Res Microbiol. 1989 Nov-Dec;140(9):615–626. doi: 10.1016/0923-2508(89)90193-9. [DOI] [PubMed] [Google Scholar]