A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid

Rodney C. Arcenas, James R. Uhl, Seanne P. Buckwalter, Andrew Harold Limper, Dana Crino, Glenn D. Roberts, Nancy L. Wengenack

Research output: Contribution to journalArticle

50 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)169-175
Number of pages7
JournalDiagnostic Microbiology and Infectious Disease
Volume54
Issue number3
DOIs
StatePublished - Mar 2006

Fingerprint

Pneumocystis
Pneumocystis carinii
Bronchoalveolar Lavage Fluid
Real-Time Polymerase Chain Reaction
Fluorescence Resonance Energy Transfer
Pneumonia
Coloring Agents
Staining and Labeling
Polymerase Chain Reaction
Antibodies
Genes

Keywords

  • LightCycler
  • PCR
  • Pneumocystis

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology
  • Microbiology
  • Parasitology
  • Virology
  • Immunology and Allergy
  • Infectious Diseases

Cite this

A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid. / Arcenas, Rodney C.; Uhl, James R.; Buckwalter, Seanne P.; Limper, Andrew Harold; Crino, Dana; Roberts, Glenn D.; Wengenack, Nancy L.

In: Diagnostic Microbiology and Infectious Disease, Vol. 54, No. 3, 03.2006, p. 169-175.

Research output: Contribution to journalArticle

Arcenas, Rodney C. ; Uhl, James R. ; Buckwalter, Seanne P. ; Limper, Andrew Harold ; Crino, Dana ; Roberts, Glenn D. ; Wengenack, Nancy L. / A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid. In: Diagnostic Microbiology and Infectious Disease. 2006 ; Vol. 54, No. 3. pp. 169-175.
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