Comparison of multiplex PCR and culture for detection of Legionellae in cooling tower water samples.
We compared multiplex polymerase chain reaction (PCR) and culture for detecting the presence of Legionella pneumophila and Legionella spp in cooling tower water samples. Multiplex PCR was performed after phenol extraction of DNA from the samples. The set of primers for the PCR assay involved the 5S rRNA (Legionella spp) and the mip (macrophage infectivity potentiator gene, specific for L. pneumophila) genes as target sequences for amplification. Both the sensitivity and the specificity of the PCR assay were 100% when the 5S rRNA gene was used as target sequence. Isolation of Legionellae from the samples was observed only with the PCR-positive samples. We propose that PCR be used as a screening test before attempting to culture Legionellae from cooling tower water samples.
Chang
H R
HR
Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore.
Loo
L H
LH
Kuah
B G
BG
Heng
B H
BH
D000389:Air Conditioning; D001483:Base Sequence; D017931:DNA Primers; D019085:Fluorescent Antibody Technique, Direct; D007875:Legionella / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012846:Singapore; D014871:Water Microbiology
D017931:DNA Primers