ArticleJournal of clinical microbiology2001
Development of conventional and real-time PCR assays for detection of Legionella DNA in respiratory specimens.
Article in Journal of clinical microbiology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers, 1 of them a synthesis that pooled it.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
31 citing papers in PubMed, 1 synthesis or guideline pooled it, 82 citations in OpenAlex.
- Diagnostic Accuracy of PCR Alone and Compared to Urinary Antigen Testing for Detection of Legionella spp.: a Systematic Review.Journal of clinical microbiology · 2016Pooled it
- How to identify respiratory pathogens in primary health care - a review on the benefits, prospects and pitfalls in using point of care tests.Infection · 2025Review
- New methods to detect bacterial or viral infections in patients with chronic obstructive pulmonary disease.Expert review of respiratory medicine · 2024Review
- Investigation of Conditions for Capture of LiveBiosensors · 2022Article
- Overview of the Clinical and Molecular Features of Legionella Pneumophila: Focus on Novel Surveillance and Diagnostic Strategies.Antibiotics (Basel, Switzerland) · 2022Review
- Water Sampling Module for Collecting and ConcentratingBiosensors · 2021Article
- Loop-mediated isothermal amplification for detection ofInfection and drug resistance · 2019Article
- Current and emerging Legionella diagnostics for laboratory and outbreak investigations.Clinical microbiology reviews · 2015Review
- Molecular Detection of Legionella: Moving on From mip.Frontiers in microbiology · 2010Article
- Direct sequencing of Legionella pneumophila from respiratory samples for sequence-based typing analysis.Journal of clinical microbiology · 2009Article
- Identification of legionella species by use of an oligonucleotide array.Journal of clinical microbiology · 2009Article
- Evaluation of a nested-PCR-derived sequence-based typing method applied directly to respiratory samples from patients with Legionnaires' disease.Journal of clinical microbiology · 2009Article
- Utility of real-time PCR for diagnosis of Legionnaires' disease in routine clinical practice.Journal of clinical microbiology · 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
- Reactive Legionella pneumophila arthritis diagnosed by polymerase chain reaction.Rheumatology international · 2007Article
- Identification and differentiation of Legionella pneumophila and Legionella spp. with real-time PCR targeting the 16S rRNA gene and species identification by mip sequencing.Applied and environmental microbiology · 2006Article
- Simultaneous detection of pathogens in clinical samples from patients with community-acquired pneumonia by real-time PCR with pathogen-specific molecular beacon probes.Journal of clinical microbiology · 2006Article
- Measurement of complement receptor 1 on neutrophils in bacterial and viral pneumonia.BMC infectious diseases · 2006Article
- Real-time PCR in clinical microbiology: applications for routine laboratory testing.Clinical microbiology reviews · 2006Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The development and validation of a PCR assay based on the use of new 16S ribosomal DNA (rDNA)-targeted primers to detect Legionella DNA in respiratory specimens are described. The assay was originally developed as conventional PCR followed by electrophoretic detection and was then adapted to Lightcycler format with SYBR Green I detection and melting curve analysis. The 73 Legionella pneumophila strains tested were amplified with both applications. In addition, 21 and 23 out of 27 other Legionella strains were found positive by conventional and real-time PCR assays, respectively, including the clinically important species L. micdadei, L. bozemaniae, and L. dumoffii. Two DNA purification methods were compared using artificially seeded clinical specimens: a standard organic extraction method and a commercial kit based on adsorption of DNA to silica particles. The detection limit of the assay varied from 2 CFU to >200,000 CFU per ml of clinical specimen, depending on the background sample (i.e., pooled sputa or BAL fluids) and the DNA purification method, the silica method achieving lower detection limits. Analysis of 77 clinical samples (66 bronchoalveolar lavage fluid and 11 sputum samples) by conventional PCR yielded results that were consistent with Legionella culture results. The melting curve analysis in the Lightcycler system readily detected the specific amplification products. However, run-to-run variations in the measured melting temperatures required normalization against the standard sample in each run. The results obtained with the clinical specimens were similar to those obtained with conventional PCR, but more samples are required to determine whether the system can be applied to routine screening of samples for the presence of Legionella DNA.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.