Abstract
A method based on PCR amplification of the 16S rRNA gene (rDNA)-23S rDNA intergenic regions was developed for the identification of species within the family Legionellaceae. The sizes of the PCR products varied from 1,353 to 350 bp. Strains of Legionella pneumophila were characterized as having products of approximately 900 and 530 bp, and L. birminghamensis had products of 1,390, 960, and 380 bp. Of the 38 species of legionellae examined, only 7 were indistinguishable (L. erythra from L. rubrilucens, L. anisa or L. cherrii from L. tucsonensis, and L. quateirensis from L. shakespearei). Two environmental isolates were identified as L. pneumophila. Strain LLAP-3, which was a symbiont of amoebae, could not be associated with any Legionella sp. studied.
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- Barry T., Colleran G., Glennon M., Dunican L. K., Gannon F. The 16s/23s ribosomal spacer region as a target for DNA probes to identify eubacteria. PCR Methods Appl. 1991 Aug;1(1):51–56. doi: 10.1101/gr.1.1.51. [DOI] [PubMed] [Google Scholar]
- Brenner D. J. Classification of Legionellaceae. Current status and remaining questions. Isr J Med Sci. 1986 Sep;22(9):620–632. [PubMed] [Google Scholar]
- Brenner D. J., Steigerwalt A. G., McDade J. E. Classification of the Legionnaires' disease bacterium: Legionella pneumophila, genus novum, species nova, of the family Legionellaceae, familia nova. Ann Intern Med. 1979 Apr;90(4):656–658. doi: 10.7326/0003-4819-90-4-656. [DOI] [PubMed] [Google Scholar]
- Feeley J. C., Gibson R. J., Gorman G. W., Langford N. C., Rasheed J. K., Mackel D. C., Baine W. B. Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila. J Clin Microbiol. 1979 Oct;10(4):437–441. doi: 10.1128/jcm.10.4.437-441.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fox A., Lau P. Y., Brown A., Morgan S. L., Zhu Z. T., Lema M., Walla M. D. Capillary gas chromatographic analysis of carbohydrates of Legionella pneumophila and other members of the family Legionellaceae. J Clin Microbiol. 1984 Mar;19(3):326–332. doi: 10.1128/jcm.19.3.326-332.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fry N. K., Rowbotham T. J., Saunders N. A., Embley T. M. Direct amplification and sequencing of the 16S ribosomal DNA of an intracellular Legionella species recovered by amoebal enrichment from the sputum of a patient with pneumonia. FEMS Microbiol Lett. 1991 Oct 1;67(2):165–168. doi: 10.1016/0378-1097(91)90348-e. [DOI] [PubMed] [Google Scholar]
- Fry N. K., Warwick S., Saunders N. A., Embley T. M. The use of 16S ribosomal RNA analyses to investigate the phylogeny of the family Legionellaceae. J Gen Microbiol. 1991 May;137(5):1215–1222. doi: 10.1099/00221287-137-5-1215. [DOI] [PubMed] [Google Scholar]
- Hookey J. V., Bryden J., Gatehouse L. The use of 16S rDNA sequence analysis to investigate the phylogeny of Leptospiraceae and related spirochaetes. J Gen Microbiol. 1993 Nov;139(11):2585–2590. doi: 10.1099/00221287-139-11-2585. [DOI] [PubMed] [Google Scholar]
- Jensen M. A., Webster J. A., Straus N. Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol. 1993 Apr;59(4):945–952. doi: 10.1128/aem.59.4.945-952.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mitchell C. D., Bean B., Gentry S. R., Groth K. E., Boen J. R., Balfour H. H., Jr Acyclovir therapy for mucocutaneous herpes simplex infections in immunocompromised patients. Lancet. 1981 Jun 27;1(8235):1389–1392. doi: 10.1016/s0140-6736(81)92569-1. [DOI] [PubMed] [Google Scholar]
- Saunders N. A., Doshi N., Harrison T. G. A second serogroup of Legionella erythra serologically indistinguishable from Legionella rubrilucens. J Appl Bacteriol. 1992 Mar;72(3):262–265. doi: 10.1111/j.1365-2672.1992.tb01833.x. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Measurement of DNA length by gel electrophoresis. Anal Biochem. 1979 Dec;100(2):319–323. doi: 10.1016/0003-2697(79)90235-5. [DOI] [PubMed] [Google Scholar]
- Springer N., Ludwig W., Drozański W., Amann R., Schleifer K. H. The phylogenetic status of Sarcobium lyticum, an obligate intracellular bacterial parasite of small amoebae. FEMS Microbiol Lett. 1992 Sep 15;75(2-3):199–202. doi: 10.1016/0378-1097(92)90403-b. [DOI] [PubMed] [Google Scholar]
- Valentine J. E., Boyle M. J., Sewell W. A., Gough N. M. Presence of single-stranded DNA in PCR products of slow electrophoretic mobility. Biotechniques. 1992 Aug;13(2):222–224. [PubMed] [Google Scholar]