Direct amplification and sequencing of the 16S ribosomal DNA of an intracellular Legionella species recovered by amoebal enrichment from the sputum of a patient with pneumonia.
DNA coding for the 16S rRNA of an intracellular bacterium was directly amplified from lysed cells of a host amoebae using the polymerase chain reaction and primers specific for eubacteria. The amoebae had been used to recover an uncultured bacterium observed in the sputum of a patient with pneumonia. The amplified DNA was sequenced directly and compared with published 16S rRNA sequences. The analysis revealed that the intracellular bacterium is a member of the genus Legionella and that it is different from species, including L. pneumophila, for which 16S ribosomal RNA sequence data are available.
Fry
N K
NK
Molecular Biology Unit, Central Public Health Laboratory, London, U.K.
Rowbotham
T J
TJ
Saunders
N A
NA
Embley
T M
TM
D000048:Acanthamoeba; D000818:Animals; D001483:Base Sequence; D004275:DNA, Ribosomal / Q000235:genetics*; D006801:Humans; D007875:Legionella / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D007876:Legionellosis / Q000382:microbiology; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D013183:Sputum / Q000382:microbiology*
D004275:DNA, Ribosomal; D009838:Oligodeoxyribonucleotides; D012336:RNA, Ribosomal, 16S