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Journal of Clinical Microbiology, January 2001, p. 162-169, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.162-169.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Rapid Identification of Clinically Relevant Legionella spp. by Analysis of Transfer DNA Intergenic Spacer Length Polymorphism

Yves De Gheldre,1,* Nicole Maes,1 François Lo Presti,2 Jerome Etienne,2 and Marc Struelens1

Laboratoire de Référence des Legionella, Service de Microbiologie, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium,1 and Centre National de Référence des Legionella, EA1655 Faculté de Médecine R. T. H. Laennec, Lyon, France2

Received 7 June 2000/Returned for modification 22 August 2000/Accepted 6 October 2000

Analysis of PCR-amplified transfer DNA (tDNA) intergenic spacers was evaluated as a rapid method for identification to the species level of 18 species of Legionella known as human pathogens. Type strains (n = 19), reference strains (n = 16), environmental strains (n = 31), and clinical strains (n = 32) were tested. PCR products using outwardly directed tDNA consensus primers were separated on polyacrylamide gels and analyzed with automated laser fluorescence. Test results were obtained in 8 h starting with 72-h-old bacterial growth on solid medium. Species-specific patterns were obtained for all 18 Legionella species tested: Legionella anisa, L. bozemanii serogroups 1 and 2, L. cincinnatiensis, L. dumoffii, L. feeleii serogroups 1 and 2, L. gormanii, L. hackeliae serogroups 1 and 2, L. jordanis, L. lansingensis, L. longbeachae serogroups 1 and 2, L. lytica, L. maceachernii, L. micdadei, L. oakridgensis, L. parisiensis, L. pneumophila serogroups 1 to 14, L. sainthelensi serogroup 2, L. tucsonensis, and L. wadsworthii. Computer-assisted matching of tDNA-intergenic length polymorphism (ILP) patterns identified all 63 environmental and clinical strains to the species level and to serogroup for some strains. tDNA-ILP analysis is proposed as a routinely applicable method which allows rapid identification of environmental and clinical isolates of Legionella spp. associated with legionellosis.

* Corresponding author. Mailing address: Laboratoire de Microbiologie, Hôpital Erasme, Université Libre de Bruxelles, 808 Route de Lennik, 1070 Brussels, Belgium. Phone: 32 2 555 4518. Fax: 32 2 555 3770. E-mail: ydegheld{at}ulb.ac.be.

Journal of Clinical Microbiology, January 2001, p. 162-169, Vol. 39, No. 1
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.1.162-169.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.