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A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid.
Diagn Microbiol Infect Dis. 2006 Mar; 54(3):169-75.DM

Abstract

Pneumocystis jiroveci is an important cause of pneumonia in immunocompromised individuals. This organism cannot be cultured, and therefore, diagnosis relies on microscopic identification of the organism using stains or antibodies. Although simple, these tests are insensitive and require expertise for accurate interpretation. We developed a real-time polymerase chain reaction (PCR) assay that provides sensitive and objective detection of Pneumocystis from bronchoalveolar lavage fluid. Primers and fluorescence resonance energy transfer probes were developed that target the cdc2 gene of P. jiroveci. Assay sensitivity is 6 copies of target per microliter of sample. No cross-reactivity occurs with other pathogens, and the PCR assay has a 21% increase in clinical sensitivity as compared with Calcofluor white staining. The real-time PCR assay provides a sensitive, rapid, and objective method for the detection of Pneumocystis from bronchoalveolar lavage fluid.

Authors+Show Affiliations

Arcenas RC
Division of Clinical Microbiology, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
Uhl JR
No affiliation info available
Buckwalter SP
No affiliation info available
Limper AH
No affiliation info available
Crino D
No affiliation info available
Roberts GD
No affiliation info available
Wengenack NL
No affiliation info available

MeSH

BenzenesulfonatesBronchoalveolar Lavage FluidDNA PrimersDNA, FungalFluorescence Resonance Energy TransferFluorescent DyesFungal ProteinsHumansPneumocystis cariniiPolymerase Chain ReactionSensitivity and SpecificityStaining and Labeling

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article

Language

eng

PubMed ID

16423488

Citation

Arcenas, Rodney C., et al. "A Real-time Polymerase Chain Reaction Assay for Detection of Pneumocystis From Bronchoalveolar Lavage Fluid." Diagnostic Microbiology and Infectious Disease, vol. 54, no. 3, 2006, pp. 169-75.
Arcenas RC, Uhl JR, Buckwalter SP, et al. A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid. Diagn Microbiol Infect Dis. 2006;54(3):169-75.
Arcenas, R. C., Uhl, J. R., Buckwalter, S. P., Limper, A. H., Crino, D., Roberts, G. D., & Wengenack, N. L. (2006). A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid. Diagnostic Microbiology and Infectious Disease, 54(3), 169-75.
Arcenas RC, et al. A Real-time Polymerase Chain Reaction Assay for Detection of Pneumocystis From Bronchoalveolar Lavage Fluid. Diagn Microbiol Infect Dis. 2006;54(3):169-75. PubMed PMID: 16423488.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid. AU - Arcenas,Rodney C, AU - Uhl,James R, AU - Buckwalter,Seanne P, AU - Limper,Andrew H, AU - Crino,Dana, AU - Roberts,Glenn D, AU - Wengenack,Nancy L, Y1 - 2006/01/19/ PY - 2005/04/12/received PY - 2005/08/08/accepted PY - 2006/1/21/pubmed PY - 2006/8/8/medline PY - 2006/1/21/entrez SP - 169 EP - 75 JF - Diagnostic microbiology and infectious disease JO - Diagn Microbiol Infect Dis VL - 54 IS - 3 N2 - Pneumocystis jiroveci is an important cause of pneumonia in immunocompromised individuals. This organism cannot be cultured, and therefore, diagnosis relies on microscopic identification of the organism using stains or antibodies. Although simple, these tests are insensitive and require expertise for accurate interpretation. We developed a real-time polymerase chain reaction (PCR) assay that provides sensitive and objective detection of Pneumocystis from bronchoalveolar lavage fluid. Primers and fluorescence resonance energy transfer probes were developed that target the cdc2 gene of P. jiroveci. Assay sensitivity is 6 copies of target per microliter of sample. No cross-reactivity occurs with other pathogens, and the PCR assay has a 21% increase in clinical sensitivity as compared with Calcofluor white staining. The real-time PCR assay provides a sensitive, rapid, and objective method for the detection of Pneumocystis from bronchoalveolar lavage fluid. SN - 0732-8893 UR - https://www.unboundmedicine.com/medline/citation/16423488/A_real_time_polymerase_chain_reaction_assay_for_detection_of_Pneumocystis_from_bronchoalveolar_lavage_fluid_ DB - PRIME DP - Unbound Medicine ER -