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RESEARCH PRODUCT
Single tube real time PCR for detection of Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila from clinical samples of CAP.
Mohammad Mehdi FeizabadiBizhan NomanpourToraj BabaeiSiroos JafariArash Ghodousisubject
Settore MED/07 - Microbiologia E Microbiologia ClinicaMycoplasma pneumoniaemedicine.disease_causeReal-Time Polymerase Chain ReactionLegionella pneumophilaSensitivity and SpecificityMicrobiologyLegionella pneumophilaCommunity-acquired pneumoniacommunity acquired pneumonia CAP real time PCR Streptococcus pneumonia Legionella pneumophila Chlamydophila pneumonia Mycoplasma pneumoniaeStreptococcus pneumoniaeMultiplex polymerase chain reactionmedicinePneumonia BacterialHumansMultiplexGeneral Immunology and MicrobiologybiologyBacteriaGeneral MedicineChlamydophila pneumoniaebiology.organism_classificationmedicine.diseaseVirologyrespiratory tract diseasesMycoplasma pneumoniaeCommunity-Acquired InfectionsReal-time polymerase chain reactionStreptococcus pneumoniaeChlamydophila pneumoniaeMultiplex Polymerase Chain Reactiondescription
We designed a multiplex real time PCR for rapid, sensitive and specific detection of Streptococcus pneumoniae, Legionella pneumophila, Chlamydophila pneumoniae and Mycoplasma pneumoniae. The study cases consisted of 129 patients with community acquired pneumonia (CAP). Bacteriological techniques were implemented for detection of the cultivable organisms. DNA were extracted from sputa, throat swabs, bronchoalveolar lavages and tracheal aspirates and used as templates in real time PCR. The primers and probes were designed for cbpA (S. pneumoniae), p1adhesin (M. pneumoniae), mip (L. pneumophila) and ompA (C. pneumoniae). After optimization of real time PCR for every organism, the experiments were continued in multiplex in a single tube. Of 129 CAP specimens, the positive cultures included 14 (10.85%) for S. pneumoniae, 9 (6.98%) for L. pneumophila and 3 (2.33%) for M. pneumoniae. Four specimens (3.10%) were positive for C. pneumoniae by real time PCR. The sensitivity of our real time PCR was 100% for all selected bacteria. The specificity of the test was 98.26%, 98.34%, 100% and 100% for S. pneumoniae, L. pneumophila, M. pneumoniae and C. pneumoniae, respectively. This is the first report on the use of multiplex real time PCR for detection of CAP patients in the Middle East. The method covers more than 90% of the bacterial pathogens causing CAP.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2012-07-04 | Acta microbiologica et immunologica Hungarica |