PCR as a test for the presence or absence of Legionella in (cooling) water.
Legionella pneumophila, a Gram-negative bacterium, is the causative agent of legionellosis. Traditionally, culture methods are normally used to detect Legionella species in different types of water (e.g. surface or tap water, circulating systems, air conditioners and their cooling devices). In this study the PCR conditions to detect Legionella were optimised based on the EnviroAmp Legionella kit (Perkin-Elmer) which is no longer commercially available. The PCR is very sensitive and specific in indicating the presence or absence (no quantification with classical PCR) of Legionella spp in general and more specifically L. pneumophila. To identify L. pneumophila. DNA sequences from the mip (macrophage infectivity potentiator) gene were amplified. The mip gene is conserved and specific for L. pneumophila although mip-like genes are also present in other Legionella spp. The PCR techniques were able to detect small amounts of Legionella in tap water samples. Cooling water, however, often contained PCR-inhibiting substances that could result in false negative PCR results for Legionella.
Declerck
P
P
Laboratory of Aquatic Ecology, Zoological Institute, Catholic University Leuven, Charles De Beriotstraat 32, B-3000 Leuven, Belgium. aquabio@bio.kuleuven.ac.be
Verelst
L
L
Duvivier
L
L
Van Damme
A
A
Ollevier
F
F
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D004784:Environmental Monitoring / Q000379:methods; D020104:Immunophilins / Q000032:analysis / Q000235:genetics*; D007875:Legionella / Q000235:genetics* / Q000302:isolation & purification; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics*; D019696:Peptidylprolyl Isomerase; D016133:Polymerase Chain Reaction; D018570:Risk Assessment; D012680:Sensitivity and Specificity; D014871:Water Microbiology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; D020104:Immunophilins; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase