COMPARISON OF DETECTION METHODS FOR LEGIONELLA SPECIES IN ENVIRONMENTAL WATER BY COLONY ISOLATION, FLUORESCENT-ANTIBODY STAINING, AND POLYMERASE CHAIN-REACTION

Three detection methods for Legionella species in water samples from cooling towers and a river were examined. Direct counting of bacteria stained with fluorescent antibody (FA) for L. pneumophila (serogroups 1 to 6) could detect the cell of 10(4) to 10(6) cell 100 ml in all 14 samples, while colony counting method detected 10 to 10(3) CFU/100 ml only in 8 samples from cooling towers. Polymerase chain reaction (PCR) assay with primers to amplify 16S ribosomal DNA sequence of most Legionella species (LEG primer) detected legionellae in 13 samples, while species-specific primers for L. pneumophila detected the DNAs from 3 samples. In laboratory examination, LEG primers could amplify DNAs of 29 species of genus Legionella with high sensitivity, even from 1 cell of L. pneumophila GIFU 9134. The PCR assay with LEG primers was specific and sensitive met.hods to be satisfied the survey of legionellae. Thus, PCR assay is a suitable method to detect and monitor Legionella species in an environment.