ArticleThe Journal of molecular diagnostics : JMD2005
Single-run, parallel detection of DNA from three pneumonia-producing bacteria by real-time polymerase chain reaction.
Article in The Journal of molecular diagnostics : JMD, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 41 citations in OpenAlex.
- Underdiagnosing of Mycoplasma pneumoniae infections as revealed by use of a respiratory multiplex PCR panel.Diagnostic microbiology and infectious disease · 2016Article
- Mycoplasma pneumoniae detection causes excess antibiotic use in Norwegian general practice: a retrospective case-control study.The British journal of general practice : the journal of the Royal College of General Practitioners · 2015Article
- Molecular detection of Mycoplasma pneumoniae by quantitative real-time PCR in patients with community acquired pneumonia.The Indian journal of medical research · 2013Article
- Molecular methods for the detection of Mycoplasma and ureaplasma infections in humans: a paper from the 2011 William Beaumont Hospital Symposium on molecular pathology.The Journal of molecular diagnostics : JMD · 2012Review
- Lower respiratory infections among hospitalized children in New Caledonia: a pilot study for the Pneumonia Etiology Research for Child Health project.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2012Article
- Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood.Protein & cell · 2012Article
- Acute respiratory infection due to Mycoplasma pneumoniae: current status of diagnostic methods.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2010Review
- Current applications and future trends of molecular diagnostics in clinical bacteriology.Analytical and bioanalytical chemistry · 2009Review
- Simultaneous detection of Chlamydophila pneumoniae and Mycoplasma pneumoniae by use of molecular beacons in a duplex real-time PCR.Journal of clinical microbiology · 2008Article
- Multiple Chlamydiaceae species in trachoma: implications for disease pathogenesis and control.PLoS medicine · 2008Article
- Currently used nucleic acid amplification tests for the detection of viruses and atypicals in acute respiratory infections.Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology · 2007Review
- Molecular typing of a Legionella pneumophila outbreak in Ontario, Canada.Journal of medical microbiology · 2007Article
- Spectrum of viruses and atypical bacteria in intercontinental air travelers with symptoms of acute respiratory infection.The Journal of infectious diseases · 2007Article
- Development of an internal positive control for rapid diagnosis of avian influenza virus infections by real-time reverse transcription-PCR with lyophilized reagents.Journal of clinical microbiology · 2006Article
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A molecular assay for parallel detection of three bacteria, Chlamydia (C.) pneumoniae, Legionella (L.) spp., and Mycoplasma (M.) pneumoniae, in clinical specimens by a set of real-time polymerase chain reactions (PCRs) in a single run was evaluated. Bacterial DNAs were extracted by an automated DNA extraction protocol on the MagNA Pure LC System. Amplification and detection were done by real-time PCR on the LightCycler (LC) instrument. For amplification, specific oligonucleotides derived from the 16s rRNA genes of C. pneumoniae, L. spp., and M. pneumoniae were used. The three assays were complemented with an internal control (IC), a specially designed DNA fragment which contains the specific primer binding sites for the three PCRs. The IC was added to the samples, co-extracted, and co-amplified. Primers and hybridization probes were designed to suit one LC PCR program. LC PCRs were established, detection limits were determined, and clinical samples were tested. The detection limits were found between 5.0 and 0.5 IFU/CFU per PCR reaction for each of the bacteria. A total number of 100 clinical specimens were tested for validation of the molecular assay. Tested samples included 63 bronchoalveolar lavages (BALs) and 37 induced sputa specimens. The internal control was detected in all negative and low-positive samples; no inhibition was found throughout the whole study. Additionally, samples underwent testing by culture for L. spp., and M. pneumoniae; for C. pneumoniae, the serological microimmunofluorescence (MIF) test was used. In conclusion, the developed set of LC PCR assays permits parallel detection of C. pneumoniae, L. spp., and M. pneumoniae in a single LC run. This molecular assay may lead to accurate and early diagnosis of pneumonia produced by these three types of bacteria. The assay proved to be suitable for the high-throughput routine diagnostic laboratory.
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