1887

Abstract

SUMMARY:

Bacteria of the family form a monophyletic group within the -subclass of . Based on comparative sequence analysis we constructed two oligonucleotide probes complementary to regions of 16S rRNA characteristic for . Probe specificities were tested by whole-cell or dot-blot hybridization against 14 serogroups of , 22 different spp. and 72 non-legionellae reference strains. Using optimized conditions both probes hybridized to all tested strains of . Probes LEG226 and LEG705 hybridized to 71% and 90% of the species tested, respectively. With the exception of none of the non-target strains showed complete sequence homology within the target molecule. In a preliminary evaluation the results of classical techniques employing selective media, immunofluorescence and the probe assay were in good accordance for routine environmental and clinical isolates. suspended in drinking water at approximately 10-10 c.f.u.ml could be rapidly detected by a combination of membrane filtration on polycarbonate filters and whole-cell hybridization. Even after incubation for 1 year a proportion of the released cells was still detectable. hybridization also facilitated visualization of spp. cells in model biofilms. A combination of hybridization and confocal laser scanning microscopy (CLSM) was used to analyse the three-dimensional arrangement of within cells of the ciliated protozoan . Whole-cell probing with 16S rRNA-targeted oligonucleotides could, in the future, complement established techniques like immunofluorescence and PCR in ecological and epidemiological studies of .

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1995-01-01
2021-08-04
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