Detection of respiratory bacterial pathogens causing atypical pneumonia by multiplex Lightmix<sup>®</sup> RT-PCR.

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State: Public
Version: Final published version
Serval ID
serval:BIB_D7A3ACFF6749
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Detection of respiratory bacterial pathogens causing atypical pneumonia by multiplex Lightmix<sup>®</sup> RT-PCR.
Journal
International journal of medical microbiology
Author(s)
Wagner K., Springer B., Imkamp F., Opota O., Greub G., Keller P.M.
ISSN
1618-0607 (Electronic)
ISSN-L
1438-4221
Publication state
Published
Issued date
04/2018
Peer-reviewed
Oui
Volume
308
Number
3
Pages
317-323
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Pneumonia is a severe infectious disease. In addition to common viruses and bacterial pathogens (e.g. Streptococcus pneumoniae), fastidious respiratory pathogens like Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella spp. can cause severe atypical pneumonia. They do not respond to penicillin derivatives, which may cause failure of antibiotic empirical therapy. The same applies for infections with B. pertussis and B. parapertussis, the cause of pertussis disease, that may present atypically and need to be treated with macrolides. Moreover, these fastidious bacteria are difficult to identify by culture or serology, and therefore often remain undetected. Thus, rapid and accurate identification of bacterial pathogens causing atypical pneumonia is crucial. We performed a retrospective method evaluation study to evaluate the diagnostic performance of the new, commercially available Lightmix <sup>®</sup> multiplex RT-PCR assay that detects these fastidious bacterial pathogens causing atypical pneumonia. In this retrospective study, 368 clinical respiratory specimens, obtained from patients suffering from atypical pneumonia that have been tested negative for the presence of common agents of pneumonia by culture and viral PCR, were investigated. These clinical specimens have been previously characterized by singleplex RT-PCR assays in our diagnostic laboratory and were used to evaluate the diagnostic performance of the respiratory multiplex Lightmix <sup>®</sup> RT-PCR. The multiplex RT-PCR displayed a limit of detection between 5 and 10 DNA copies for different in-panel organisms and showed identical performance characteristics with respect to specificity and sensitivity as in-house singleplex RT-PCRs for pathogen detection. The Lightmix <sup>®</sup> multiplex RT-PCR assay represents a low-cost, time-saving and accurate diagnostic tool with high throughput potential. The time-to-result using an automated DNA extraction device for respiratory specimens followed by multiplex RT-PCR detection was below 4 h, which is expected to significantly improve diagnostics for atypical pneumonia-associated bacterial pathogens.
Keywords
Adolescent, Bacteria/genetics, Bacteria/isolation & purification, Bacteria/pathogenicity, Chlamydophila pneumoniae/genetics, Chlamydophila pneumoniae/isolation & purification, Chlamydophila pneumoniae/pathogenicity, DNA, Bacterial/genetics, Female, High-Throughput Screening Assays/methods, Humans, Legionella/genetics, Legionella/isolation & purification, Legionella/pathogenicity, Male, Molecular Diagnostic Techniques/methods, Multiplex Polymerase Chain Reaction/economics, Multiplex Polymerase Chain Reaction/methods, Mycoplasma pneumoniae/genetics, Mycoplasma pneumoniae/isolation & purification, Mycoplasma pneumoniae/pathogenicity, Pneumonia, Bacterial/diagnosis, Pneumonia, Bacterial/microbiology, Pneumonia, Mycoplasma/diagnosis, Pneumonia, Mycoplasma/microbiology, Reagent Kits, Diagnostic, Respiratory Tract Infections/diagnosis, Respiratory Tract Infections/microbiology, Retrospective Studies, Sensitivity and Specificity, Streptococcus pneumoniae/genetics, Streptococcus pneumoniae/isolation & purification, Atypical pneumonia, High throughput screening, Lightmix(®) kit, Molecular detection, Multiplex RT-PCR, Respiratory pathogens
Pubmed
Web of science
Open Access
Yes
Create date
08/02/2018 18:30
Last modification date
20/08/2019 15:57
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