Pneumocystis jirovecii testing by real-time polymerase chain reaction and direct examination among immunocompetent and immunosuppressed patient groups and correlation to disease specificity

John W. Wilson, Andrew Harold Limper, Thomas Grys, Theresa Karre, Nancy L. Wengenack, Matthew J. Binnicker

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

We used real-time polymerase chain reaction (PCR) targeting the cdc2 gene and direct fluorescent microscopy examination (DFME) to evaluate the prevalence of Pneumocystis jirovecii among immunocompetent patients without clinical pulmonary infection and immunosuppressed patients evaluated for opportunistic pulmonary infections. Among 102 bronchoalveolar lavage samples collected from immunocompetent patients without infection, none tested positive for P. jirovecii by either DFME or real-time PCR despite the presence of other comorbidities. Among patients with suspected pulmonary infection and tested with either assay, real-time PCR produced a higher number of positive results compared to DFME and increased P. jirovecii detection by 7% when added to DFME-negative samples. Real-time PCR may have increased sensitivity for P. jirovecii detection over DFME and decrease the risk of sample contamination compared to conventional and nested PCR. The use of single-copy gene targets (e.g., cdc2) may lower the rate of "colonization" detection and confer a high predictive value for Pneumocystis pneumonia.

Original languageEnglish (US)
Pages (from-to)145-152
Number of pages8
JournalDiagnostic Microbiology and Infectious Disease
Volume69
Issue number2
DOIs
StatePublished - Feb 2011

Fingerprint

Pneumocystis carinii
Real-Time Polymerase Chain Reaction
Microscopy
Lung
Infection
Pneumocystis Pneumonia
Gene Targeting
Opportunistic Infections
Bronchoalveolar Lavage
Comorbidity
Polymerase Chain Reaction
Genes

Keywords

  • P. jirovecii
  • Pneumocystis pneumonia
  • Real-time PCR, Direct examination

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

Cite this

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abstract = "We used real-time polymerase chain reaction (PCR) targeting the cdc2 gene and direct fluorescent microscopy examination (DFME) to evaluate the prevalence of Pneumocystis jirovecii among immunocompetent patients without clinical pulmonary infection and immunosuppressed patients evaluated for opportunistic pulmonary infections. Among 102 bronchoalveolar lavage samples collected from immunocompetent patients without infection, none tested positive for P. jirovecii by either DFME or real-time PCR despite the presence of other comorbidities. Among patients with suspected pulmonary infection and tested with either assay, real-time PCR produced a higher number of positive results compared to DFME and increased P. jirovecii detection by 7{\%} when added to DFME-negative samples. Real-time PCR may have increased sensitivity for P. jirovecii detection over DFME and decrease the risk of sample contamination compared to conventional and nested PCR. The use of single-copy gene targets (e.g., cdc2) may lower the rate of {"}colonization{"} detection and confer a high predictive value for Pneumocystis pneumonia.",
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