Use of the Polymerase Chain Reaction to Detect Legionella DNA in Urine and Serum Samples from Patients with Pneumonia

Abstract

Legionella pneumonia can be difficult to diagnose. Existing laboratory tests for detecting Legionella species lack sensitivity or provide only a retrospective diagnosis. We used the polymerase chain reaction (PCR) with primers that amplify a 104-base pair segment of the coding region of the 5S tRNA gene to detect Legionella DNA in urine and serum samples from patients with pneumonia. Stored urine and serum samples from patients enrolled in two prospective studies of pneumonia were tested. Legionella DNA was detected in urine and/or serum samples from 18 (64%) of 28 patients with legionella pneumonia diagnosed by conventional tests, but it was not detected in urine or serum samples from 24 patients with pneumonia due to other organisms. The sensitivity of PCR improved to 73% if testing was restricted to samples taken within 4 days of the onset of symptoms. Detection of Legionella DNA in urine and serum promises to be a valuable tool for the rapid diagnosis of legionella pneumonia.

Laboratory diagnosis of legionnaires’ disease due to Legionella pneumophila serogroup 1: comparison of phenotypic and genotypic methods

“…In this study, the overall sensitivity of the 5S rRNA PCR was 63 % of the total sera tested. Legionellaspecific DNA has previously been detected in serum, but with low sensitivities of 30 and 43 % (Murdoch et al, 1996;Matsiota-Bernard et al, 1997). As seen in this study and others (reviewed by Murdoch, 2003b), these sensitivity levels can be increased if samples are obtained early in the course of the illness and when multiple samples are tested.…”

Development and Clinical Evaluation of an Internally Controlled, Single-Tube Multiplex Real-Time PCR Assay for Detection of Legionella pneumophila and Other Legionella Species

“…Therefore, clinically the identification of the actual Legionella species is required. Diagnostic PCR assays have principally targeted specific regions within 16S rRNA genes (rDNA) (3,9,12,21,23,26), 5S rDNA (9), or the macrophage inhibitor potentiator (mip) gene (1,9,15,16,17,20,22,26). Real-time PCR has added benefits to routine diagnosis, as it minimizes manual time for the PCR and negates the use of post-PCR analysis.…”

Serological Survey of Brucellosis in Camel of Gujarat

“…Several studies have documented the detection of Brucella specific genome from the serum samples using PCR (Shome et al, 2013;Brown et al, 1995;Murdoch et al, 1996;Bougnoux et al, 1999;Kawamura et al, 1999). The use of serum instead of wholeblood samples offers several advantages for nucleic acid amplification.…”