A Legionella-specific DNA probe detects organisms in lung tissue homogenates from intranasally inoculated mice.
We have prepared a DNA probe from internal sequences of the gene encoding the Legionella pneumophila major outer membrane protein (MOMP). Immunologic studies of the MOMP have confirmed that it possesses both genus-specific and species-specific antigenic domains, but possesses no cross-reactivity with non-Legionella species. At the DNA levels, the 3' half of the gene contains sequences that are homologous to DNA from all strains tested within the genus, whereas the 5' half of the gene has homology with L. pneumophila strains only. Homology of the gene with non-legionellae has not been detected even under low stringency conditions. To test the utility of this probe for detecting organisms in tissue, we tested crude homogenates of mouse lungs representing 1/1,000th of the total lung mass. After intranasal inoculation with 2 X 10(8) colony-forming units of L. pneumophila, mice were sacrificed at various intervals (10 mice per group). Since L. pneumophila does not produce a propagating infection in these animals, cultures of lung tissue from successive days after inoculation showed a roughly linear decline in viable L. pneumophila (total lung yield: 10(8) on Day 0, 5 X 10(7) on Day 2, 10(5) on Day 5, 10(3) on Day 9, and less than 10(2) on Day 15). By DNA dot hybridization with the MOMP probe, we detected positive signals from most animals on Days 0 and 2, suggesting a threshold sensitivity of between 50,000 and 100,000 organisms with our current methods. Advances in DNA probe technology may soon permit the rapid, specific identification of either L. pneumophila or other Legionella species in pathologic specimens.
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial; D007875:Legionella / Q000235:genetics*; D007877:Legionnaires' Disease / Q000175:diagnosis*; D008168:Lung / Q000382:microbiology*; D051379:Mice; D008819:Mice, Nude; D009693:Nucleic Acid Hybridization
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial