Rapid identification of clinically relevant Legionella spp. by analysis of transfer DNA intergenic spacer length polymorphism.
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.
De Gheldre
Y
Y
Laboratoire de Référence des Legionella, Service de Microbiologie, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium. ydegheld@ulb.ac.be
Maes
N
N
Presti
F L
FL
Etienne
J
J
Struelens
M
M
D019295:Computational Biology / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics*; D004783:Environmental Microbiology; D006801:Humans; D007875:Legionella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D007876:Legionellosis / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012343:RNA, Transfer / Q000235:genetics*; D012015:Reference Standards; D015203:Reproducibility of Results
D004269:DNA, Bacterial; D021901:DNA, Intergenic; D012343:RNA, Transfer