Abstract
A detection system forLegionella DNA in urine samples based on the polymerase chain reaction (PCR) was developed and tested on infected guinea pigs and patients suffering from pneumonia. Results were compared with standard methods for diagnosis of Legionnaires' disease. A primer system was selected which amplifies a 108 bp DNA fragment of the 5S rRNA gene. The sensitivity of the PCR system was one femtogram of extractedLegionella DNA. Three methods were tested for pretreatment of urine samples. Of these, the Geneclean II kit (Bio 101, USA) gave the best results for artificially contaminated urine samples as well as those from infected guinea pigs or patients. Thirty-seven urine samples from 15 guinea pigs intraperitoneally infected with eitherLegionella pneumophila serogroup 1, 3 and 6 orLegionella micdadei, 26 urine samples of 21 patients suffering from pneumonia, and 30 control samples of patients with urinary tract infection (UTI) were tested.Legionella DNA was detected in 29 of the guinea pig urine samples; whereas, urinary antigen detection using EIA was positive in only 20 of the samples. PCR was also positive in the samples of 11 patients with pneumonia, 9 of which were confirmed by other microbiological methods, such as culture, direct fluorescent antibody test, urinary antigen detection and antibody testing. However, of the 30 control samples from patients with UTI, three samples yielded positive results. The results demonstrate thatLegionella DNA is excreted in the urine of infected individuals and that the PCR shows a higher degree of sensitivity than EIA to the detection of solubleLegionella antigen in urine. Although the positive results obtained from control samples are a potential limitation of the test, these preliminary data suggest that PCR can be a sensitive method for the diagnosis of legionellosis from urine samples.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Fliermans CB, Cherry WB, Orrison LH, Thacker L: Isolation ofLegionella pneumophila from nonepidemic-related aquatic habitats. Applied and Environmental Microbiology 1979, 37: 1239–1242.
Fliermans CB, Cherry WB, Orrison LH, Smith SJ, Tison DL, Pope DH: Ecological distribution ofLegionella pneumophila. Applied and Environmental Microbiology 1981, 41: 9–16.
Morris GK, Patton CM, Feeley JC, Johnson SE, Gorman G, Martin WT, Skaliy P, Mallison GF, Politi BD, Mackel DC: Isolation of the Legionnaires' disease bacterium from environmental samples. Annals of Internal Medicine 1979, 90: 664–666.
Tobin JO'H, Dunnill MS, French M, Morris PJ, Beare J, Fisher-Hoch S, Mitchell RG, Muers MF: Legionnaires' disease in a transplant unit: isolation of the causative agent from shower baths. Lancet, London 1980, ii: 118–121.
Dondero TJ, Rendtorff RC, Mallison GF, Weeks RM, Levy JS, Wong EW, Schaffner W: An outbreak of Legionnaires' disease associated with contaminated air conditioning cooling tower. New England Journal of Medicine 1980, 302: 365–370.
Breiman RF, Cozen W, Fields BS, Mastro TD, Carr SJ, Spika JS, and Mascola L: Role of air-sampling in an investigation of an outbreak of Legionnaires' disease associated with exposure to aerosols from an evaporative condenser. Journal of Infectious Diseases 1990, 161: 1257–1261.
Stout JE, Yu VL, Muraca P, Joly J, Troup N, Tompkins LS: Potable water as a cause of sporadic cases of community-acquired Legionnaires' disease. New England Journal of Medicine 1992, 326: 151–155.
Blatt SP, Parkinson MD, Pace E, Hoffman P, Dolan D, Lauderdale P, Zajac RA, Melcher GP: Nosocomial Legionnaires' disease: aspiration as a primary mode of transmission. American Journal of Medicine 1993, 95: 16–22.
Yu VL: Could aspiration be the major mode of transmission forLegionella? American Journal of Medicine 1993, 95: 13–15.
England AC, Fraser DW: Sporadic and epidemic nosocomial legionellosis in the United States. Epidemiologic features. American Journal of Medicine 1981, 70: 707–711.
Harrison TG, Taylor AG: A laboratory manual forLegionella. John Wiley, London, 1988.
Edelstein PH: Legionnaires' disease. Clinical Infectious Diseases 1993, 16: 741–747.
Harrison TG, Dournon E, Taylor AG: Evaluation of sensitivity of two serological tests for diagnosing pneumonia caused byLegionella pneumophila serogroup 1. Journal of Clinical Pathology 1987, 40: 77–82.
Aguero-Rosenfeld ME, Edelstein PH: Retrospective evaluation of the Du Pont radioimmunoassay kit for detection ofLegionella pneumophila serogroup 1 antigenuria in humans. Journal of Clinical Microbiology 1988, 26: 1775–1778.
Starnbach MN, Falkow S, Tompkins LS: Species-specific detection ofLegionella pneumophila in water by DNA amplification and hybridization. Journal of Clinical Microbiology 1989, 27: 1257–1261.
Mahbubani MH, Bej AK, Miller R, Haff L, DiCesare J, Atlas RM: Detection ofLegionella with polymerase chain reaction and gene probe methods. Molecular and Cellular Probes 1990, 4: 175–187.
Jaulhac B, Nowicki M, Bornstein N, Meunier O, Prevost G, Piemont Y, Fleurette J, Monteil H: Detection ofLegionella spp. in bronchoalveolar lavage fluids by DNA amplification. Journal of Clinical Microbiology 1992, 30: 920–924.
Koide M, Saito A, Kusano N, Higa F: Detection ofLegionella spp. in cooling tower water by the polymerase chain reaction method. Applied and Environmental Microbiology 1993, 59: 1943–1946.
Maiwald M, Kissel K, Srimuang S, von Knebel Doeberitz M, Sonntag HG: Comparison of polymerase chain reaction and conventional culture for the detection of legionellas in hospital water samples. Journal of Applied Bacteriology 1994, 76: 216–225.
Kessler HH, Reinthaler FF, Pschaid A, Pierer K, Kleinhappl B, Eber E, Marth E: Rapid detection ofLegionella species in bronchoalveolar lavage fluids with the EnviroAmpLegionella PCR amplification and detection kit. Journal of Clinical Microbiology 1993, 31: 3325–3328.
Edelstein PH: Improved semiselective medium for isolation ofLegionella pneumophila from contaminated clinical and environmental specimens. Journal of Clinical Microbiology 1981, 14: 298–303.
Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: a laboratory manual. 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989.
Gosting LH, Cabrian K, Sturge JC, Goldstein LC: Identification of a species-specific antigen inLegionella pneumophila by a monoclonal antibody. Journal of Clinical Microbiology 1984, 20: 1031–1035.
Helbig JH, Lück PC, Witzleb W: Diagnostik vonLegionella-Pneumonien durch Nachweis der Antigenurie mittels Enzymimmunoassay unter Verwendung von 6 unterschiedlichen Antikörperspezifitäten. Zeitschrift für die gesamte Hygiene 1989, 35: 591–593.
Helbig JH, Lück PC, Pilz C, Witzleb W: Common epitope on urinary antigen derived from differentLegionella pneumophila serogroup 1 strains recognized by a monoclonal antibody. Zentralblatt für Bakteriologie 1990, 273: 478–480.
Rajakumar R, Lacey CJN, Evans EGV, Carney JA: Use of slide agglutination test for rapid diagnosis of vaginal candidosis. Genitourinary Medicine 1987, 63: 192–195.
Matsiota-Bernard P, Pitsouni E, Legakis N, Nauciel C: Evaluation of commercial amplification kit for detection ofLegionella pneumophila in clinical specimens. Journal of Clinical Microbiology 1994, 32: 1503–1505.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Maiwald, M., Schill, M., Stockinger, C. et al. Detection ofLegionella DNA in human and guinea pig urine samples by the polymerase chain reaction. Eur. J. Clin. Microbiol. Infect. Dis. 14, 25–33 (1995). https://doi.org/10.1007/BF02112614
Issue Date:
DOI: https://doi.org/10.1007/BF02112614