Real-time PCR assay targets the 23S-5S spacer for direct detection and differentiation of Legionella spp. and Legionella pneumophila.
A real-time PCR for the ABI Prism 7000 system targeting the 23S-5S spacer of Legionella spp. was developed. Simultaneous detection and differentiation of Legionella spp. and Legionella pneumophila within 90 min and without post-PCR melting-curve analysis was achieved using two TaqMan probes. In sputum samples from 23 controls and 17 patients with legionellosis, defined by positive culture, urinary antigen testing, or seroconversion, 94% sensitivity and 100% specificity were observed.
Herpers
Bjorn L
BL
Department of Medical Microbiology and Immunology, St. Antonius Hospital, Koekoekslaan 1, 3435 CM Nieuwegein, The Netherlands. b.herpers@antonius.net
de Jongh
Bartelt M
BM
van der Zwaluw
Kim
K
van Hannen
Erik J
EJ
D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000032:analysis*; D006801:Humans; D007875:Legionella / Q000145:classification* / Q000235:genetics; D016952:Legionella pneumophila / Q000145:classification* / Q000235:genetics; D007877:Legionnaires' Disease / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D012341:RNA, Ribosomal, 5S / Q000235:genetics*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology
D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D012338:RNA, Ribosomal, 23S; D012341:RNA, Ribosomal, 5S