title: Comparison of detection methods for Legionella species in environmental water by colony isolation, fluorescent antibody staining, and polymerase chain reaction.
abstract: 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 methods to be satisfied the survey of legionellae. Thus, PCR assay is a suitable method to detect and monitor Legionella species in an environment.
authors.affiliation: Department of Microbiology, Gifu University School of Medicine, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: T
authors.initials: T
authors.lastname: Ezaki
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology; D001419:Bacteria / Q000302:isolation & purification; D001431:Bacteriological Techniques; D001483:Base Sequence; D004269:DNA, Bacterial / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D007875:Legionella / Q000145:classification / Q000276:immunology / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014871:Water Microbiology
chemical_list: D000907:Antibodies, Bacterial; D004269:DNA, Bacterial