title: Development of a multiplex real-time quantitative PCR assay to detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae in respiratory tract secretions.
abstract: Atypical pathogens such as Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae are an important cause of community-acquired pneumonia. The available detection methods (culture and serology) either lack sensitivity or give only a retrospective diagnosis. In order to improve their detection and quantification in respiratory samples, a real-time multiplex PCR, performed in two separate reactions, was developed for these three pathogens. The comparison of multiplex real-time and conventional PCR assay on 73 respiratory specimens showed an overall agreement of 98.3%, corresponding to 95.8%, 100% and 100% agreement for C. pneumoniae, L. pneumophila and M. pneumoniae, respectively. Clinical application of this multiplex real-time PCR was done on 40 respiratory samples from 38 patients with respiratory tract infections. Of 19 serology-positive patients, 14 were confirmed by the multiplex real-time PCR to be infected by either one of the three pathogens. All samples from serology-negative patients were negative with the multiplex real-time PCR.
authors.affiliation: Institute of Microbiology, University Hospital of Lausanne, Lausanne, Switzerland.
authors.forename: Martine
authors.initials: M
authors.lastname: Welti
authors.forename: Katia
authors.initials: K
authors.lastname: Jaton
authors.forename: Martin
authors.initials: M
authors.lastname: Altwegg
authors.forename: Roland
authors.initials: R
authors.lastname: Sahli
authors.forename: Aline
authors.initials: A
authors.lastname: Wenger
authors.forename: Jacques
authors.initials: J
authors.lastname: Bille
mesh_terms: D001709:Biotechnology; D023521:Chlamydophila Infections / Q000175:diagnosis; D016993:Chlamydophila pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics / Q000302:isolation & purification*; D007877:Legionnaires' Disease / Q000175:diagnosis; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D018410:Pneumonia, Bacterial / Q000175:diagnosis*; D011019:Pneumonia, Mycoplasma / Q000175:diagnosis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012137:Respiratory System / Q000378:metabolism / Q000382:microbiology; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial