title: Detection of Legionella by PCR in respiratory specimens using a commercially available kit.
abstract: Using polymerase chain reaction (PCR) for the detection of pathogens that are difficult to grow, such as Legionella species, may reduce difficulties encountered with culture and immunofluorescent staining. We evaluated a commercial PCR and hybridization kit, designed for environmental samples, for the detection of Legionella in respiratory specimens. Sixteen Legionella species cultures tested positive with the Perkin Elmer Legionella EnviroAmp Amplification and Detection kits (Perkin Elmer, Foster City, Calif). The assay detected as few as 100 colony-forming units per milliliter of spiked bronchoalveolar lavage (BAL) fluid, and no false-negative results were obtained. PCR inhibition by blood in the specimens was removed by washing pelleted specimens in sterile distilled water. Of 126 specimens screened with the kit, 1 induced sputum and 3 BAL specimens were positive by PCR. All 4 were validated as true-positive results by culture or serologic testing. The entire PCR and hybridization assay can be completed in less than 6 hours, whereas isolation and identification by culture requires up to 12 days, and serologic conversion may not be demonstrated for weeks. Molecular techniques based on direct extraction and amplification of DNA from respiratory specimens nay be useful for the timely diagnosis of legionellosis.
authors.affiliation: Microbiology Service, Clinical Pathology Department, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: S C
authors.initials: SC
authors.lastname: Weir
authors.forename: S H
authors.initials: SH
authors.lastname: Fischer
authors.forename: F
authors.initials: F
authors.lastname: Stock
authors.forename: V J
authors.initials: VJ
authors.lastname: Gill
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D004269:DNA, Bacterial / Q000032:analysis*; D005069:Evaluation Studies as Topic; D005188:False Negative Reactions; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics / Q000302:isolation & purification*; D007877:Legionnaires' Disease / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic