Abstract
A sensitive detection system specific for Legionella pneumophila in water was developed. This system is based on amplification of a chromosomal DNA sequence from L. pneumophila by the polymerase chain reaction, followed by detection of the amplified product by hybridization of a radiolabeled oligodeoxynucleotide. After 35 cycles of amplification, a water sample which had been seeded with 35 CFU of L. pneumophila contained sufficient amplified DNA to be detected on dot blots. Bacteria of other genera tested did not generate positive signals under these conditions. Application of this technique to environmental water samples may help identify the natural reservoirs of nosocomial and community-acquired L. pneumophila infections.
MeSH Terms
Blotting, Southern
DNA Probes
DNA, Bacterial/analysis
Electrophoresis, Agar Gel
Feces/microbiology
Gene Amplification
Humans
Legionella/genetics,isolation & purification
Nucleic Acid Hybridization
Species Specificity
Water Microbiology
Chemicals
DNA Probes
DNA, Bacterial
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Starnbach M N
Department of Microbiology and Immunology, Stanford University School of Medicine, California 94305.
Falkow S
Tompkins L S
References (11)
11 references, click to expand
-
Taxonomy and epidemiology of gram-negative bacterial plasmids studied by DNA-DNA filter hybridization in formamide.
J Gen Microbiol. 1978 Feb;104(2):269-76
PMID: 632804
-
Legionnaires' disease in a transplant unit: isolation of the causative agent from shower baths.
Lancet. 1980 Jul 19;2(8186):118-21
PMID: 6105294
-
Ubiquitousness of Legionella pneumophila in the water supply of a hospital with endemic Legionnaires' disease.
N Engl J Med. 1982 Feb 25;306(8):466-8
PMID: 7057845
-
Epidemiologic assessment of methods of transmission of legionellosis.
Zentralbl Bakteriol Mikrobiol Hyg A. 1983 Jul;255(1):52-7
PMID: 6637235
-
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.
Science. 1985 Dec 20;230(4732):1350-4
PMID: 2999980
-
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
Science. 1988 Jan 29;239(4839):487-91
PMID: 2448875
-
A Legionella-specific DNA probe detects organisms in lung tissue homogenates from intranasally inoculated mice.
Isr J Med Sci. 1986 Oct;22(10):703-5
PMID: 3793430
-
Retrospective study of Gen-Probe rapid diagnostic system for detection of legionellae in frozen clinical respiratory tract samples.
J Clin Microbiol. 1987 Jun;25(6):1022-6
PMID: 2439535
-
Electrophoresis in agarose and acrylamide gels.
Methods Enzymol. 1987;152:61-87
PMID: 2443811
-
Molecular epidemiology of Legionella species by restriction endonuclease and alloenzyme analysis.
J Clin Microbiol. 1987 Oct;25(10):1875-80
PMID: 2822760
-
Evaluation of the Gen-Probe DNA probe for the detection of legionellae in culture.
J Clin Microbiol. 1986 Mar;23(3):481-4
PMID: 2420820