Species-specific detection of Legionella using polymerase chain reaction and reverse dot-blotting.
Legionella pneumophila and some other Legionella species are capable of causing Legionnaire's disease, a potentially fatal pneumonia. The identification of legionellae by standard laboratory techniques such as culture is difficult and time consuming. In the present work, the DNA sequence of the 23S-5S spacer region was determined for 43 Legionella isolates, and the sequence information was used to develop a species-specific detection system using PCR and reverse dot-blotting which employs just one PCR amplicon to perform genus- and species-specific detection. L. pneumophila serogroups 1-16 as well as 21 non-pneumophila isolates could be identified and differentiated at the species level using this system.
Robinson
P N
PN
Neurochemistry Research Laboratory, Klinikum Benjamin-Franklin, Freien Universität Berlin, Germany.
Heidrich
B
B
Tiecke
F
F
Fehrenbach
F J
FJ
Rolfs
A
A
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D007875:Legionella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D007877:Legionnaires' Disease / Q000175:diagnosis / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; D012341:RNA, Ribosomal, 5S