ArticleJournal of clinical microbiology1997
Rapid detection of Pneumocystis carinii in bronchoalveolar lavage specimens from human immunodeficiency virus-infected patients: use of a simple DNA extraction procedure and nested PCR.
Article in Journal of clinical microbiology, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers, 2 of them syntheses that pooled it.
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Who cites it
23 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- Polymerase Chain Reaction on Respiratory Tract Specimens of Immunocompromised Patients to Diagnose Pneumocystis Pneumonia: A Systematic Review and Meta-analysis.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2024Pooled it
- The prevalence of laboratory-confirmed Pneumocystis jirovecii in HIV-infected adults in Africa: A systematic review and meta-analysis.Medical mycology · 2021Pooled it
- Performance of a Real Time PCR forJournal of fungi (Basel, Switzerland) · 2022Article
- Pathogenic microorganism detection in AIDS patients using bronchoalveolar lavage fluid.International journal of clinical and experimental pathology · 2020Article
- Role of Pneumocystis jirovecii in patients with different pulmonary underlying condition using a nested-PCR.Revista espanola de quimioterapia : publicacion oficial de la Sociedad Espanola de Quimioterapia · 2018Article
- Prevalence of colonization and mitochondrial large subunit rRNA mutation ofIranian journal of microbiology · 2016Article
- Use of nasopharyngeal aspirate for diagnosis of pneumocystis pneumonia.Journal of clinical microbiology · 2013Article
- Aetiology of pulmonary symptoms in HIV-infected smear negative recurrent PTB suspects in Kampala, Uganda: a cross-sectional study.PloS one · 2013Article
- Molecular detection of pneumocystis jirovecii in patients with respiratory tract infections.North American journal of medical sciences · 2012Article
- Colonization by Pneumocystis jirovecii and its role in disease.Clinical microbiology reviews · 2012Review
- Comparison of DNA extraction methods for microbial community profiling with an application to pediatric bronchoalveolar lavage samples.PloS one · 2012Article
- Article
- Pneumocystis colonization in immunocompetent and simian immunodeficiency virus-infected cynomolgus macaques.The Journal of infectious diseases · 2009Article
- FlindersTechnology Associates (FTA) filter paper-based DNA extraction with polymerase chain reaction (PCR) for detection of Pneumocystis jirovecii from respiratory specimens of immunocompromised patients.Journal of clinical laboratory analysis · 2007Article
- Pneumocystis pneumonia in an alcoholic patient with prolonged mechanical ventilation.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2005Article
- Molecular evidence of interhuman transmission of Pneumocystis pneumonia among renal transplant recipients hospitalized with HIV-infected patients.Emerging infectious diseases · 2004Article
- Detection of Pneumocystis carinii and characterization of mutations associated with sulfa resistance in bronchoalveolar lavage samples from human immunodeficiency virus-infected subjects.Journal of clinical microbiology · 2003Article
- Pneumocystis jiroveci internal transcribed spacer types in patients colonized by the fungus and in patients with pneumocystosis from the same French geographic region.Journal of clinical microbiology · 2003Article
- Detection of Pneumocystis DNA in samples from patients suspected of bacterial pneumonia--a case-control study.BMC infectious diseases · 2002Article
- Development and evaluation of a quantitative, touch-down, real-time PCR assay for diagnosing Pneumocystis carinii pneumonia.Journal of clinical microbiology · 2002Article
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11 authors.
Funding
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Abstract
We report on the development of a rapid nested PCR protocol for the detection of Pneumocystis carinii DNA in bronchoalveolar lavage (BAL) specimens in which the protocol included the use of a commercially available DNA extraction kit (GeneReleaser). GeneReleaser enabled us to obtain amplification-ready DNA within 20 min without requiring the purification of the DNA. The nested PCR was performed with the primers pAZ102-E, pAZ102-H, and pAZ102-L2 (A. E. Wakefield, F. J. Pixley, S. Banerji, K. Sinclair, R. F. Miller, E. R. Moxon, and J. M. Hopkin, Lancet 336:451-453, 1990.). Results were obtained in about 4 h with the adoption of denaturation, annealing, and extension steps shortened to 20 seconds. The sensitivity of the nested PCR was tested with a P. carinii cyst suspension and was found to be less than one cyst (one to eight nuclei). The detection limit was the same with the use of GeneReleaser or proteinase K-phenol chloroform for DNA extraction. The nested PCR assay was prospectively compared with staining with Giemsa and methenamine silver stains for the detection of P. carinii in 127 BAL samples from 105 human immunodeficiency virus-infected patients investigated for acute respiratory illness. Twenty-five BAL specimens (20%) were positive by staining and the nested PCR and 25 (20%) were negative by staining and positive by the nested PCR. These 25 BAL specimens with conflicting results were obtained from 23 patients, 82% of whom were receiving prophylactic therapy against P. carinii pneumonia (PCP). Only two patients were diagnosed with possible PCP. The final diagnosis was not PCP for 20 patients who were considered to be colonized or to have a low level of infection. This colonization is not of clinical importance but is of epidemiological importance. Our rapid, simple, and sensitive amplification protocol may be performed in clinical laboratories for the routine diagnosis of PCP with BAL specimens.
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