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 H. Limper, Dana Crino, Glenn D. Roberts, Nancy L. Wengenack

Research output: Contribution to journalArticle

57 Scopus citations

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 1 2006

Keywords

  • LightCycler
  • PCR
  • Pneumocystis

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

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