title: Evaluation of real-time PCR for the early detection of Legionella pneumophila DNA in serum samples.
abstract: Legionella pneumonia can be difficult to diagnose. Existing laboratory tests all have shortcomings, especially in the ability to diagnose Legionnaires' disease (LD) at an early stage of the disease in a specimen that is readily obtainable. The aim of this study was to assess the performance of PCR as a rapid diagnostic method and to compare the results of different PCR assays of serum samples from patients with LD. Samples included 151 serum samples from 68 patients with proven LD and 60 serum samples from 36 patients with respiratory tract infections other than Legionella. PCR assays were based on the 5S rRNA gene, 16S rRNA gene and the mip gene. The samples from patients with infections caused by pathogens other than Legionella all tested negative in PCR. Among the patients with proven LD 54.4 % (37/68) tested positive in 5S rRNA PCR, 52.9 % (36/68) in mip gene PCR and 30.9 % (21/68) in 16S rRNA PCR in the first available serum sample. The association between threshold cycle value in 5S PCR positive serum samples (n=49) and C-reactive protein value was determined, and showed a strong negative correlation (Pearson correlation coefficient r=-0.63, P<0.0001). In addition to existing tests for the diagnosis of LD, detection of Legionella DNA in serum could be a useful tool for early diagnosis of LD caused by any Legionella species and serogroup, and has the potential to provide a diagnosis in a time frame that could affect initial infection management.
authors.affiliation: Laboratory of Medical Microbiology and Immunology, St Elisabeth Hospital, PO Box 747, 5000 AS Tilburg, The Netherlands.
authors.forename: Bram M W
authors.initials: BMW
authors.lastname: Diederen
authors.affiliation: Laboratory of Medical Microbiology and Immunology, St Elisabeth Hospital, PO Box 747, 5000 AS Tilburg, The Netherlands.
authors.forename: Caroline M A
authors.initials: CMA
authors.lastname: de Jong
authors.affiliation: Laboratory of Medical Microbiology and Immunology, St Elisabeth Hospital, PO Box 747, 5000 AS Tilburg, The Netherlands.
authors.forename: Faïçal
authors.initials: F
authors.lastname: Marmouk
authors.affiliation: Laboratory of Microbiology and Infection Control, Amphia Hospital, PO Box 90158, 4800 RK Breda, The Netherlands.
authors.forename: Jan A J W
authors.initials: JAJW
authors.lastname: Kluytmans
authors.affiliation: Laboratory of Medical Microbiology and Immunology, St Elisabeth Hospital, PO Box 747, 5000 AS Tilburg, The Netherlands.
authors.forename: Marcel F
authors.initials: MF
authors.lastname: Peeters
authors.affiliation: Laboratory of Medical Microbiology and Immunology, St Elisabeth Hospital, PO Box 747, 5000 AS Tilburg, The Netherlands.
authors.forename: Anneke
authors.initials: A
authors.lastname: Van der Zee
mesh_terms: D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis* / Q000097:blood / Q000235:genetics; D005260:Female; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics*; D007877:Legionnaires' Disease / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000592:standards; D019696:Peptidylprolyl Isomerase / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012341:RNA, Ribosomal, 5S; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase