Sensitivity and specificity of nested and real-time PCR for the detection of Pneumocystis jiroveci in clinical specimens

Míriam J. Alvarez-Martínez, José M. Miró, Maria Eugenia Valls, Asunción Moreno, Paula V. Rivas, Manel Solé, Natividad Benito, Pere Domingo, Carmen Muñoz, Esteban Rivera, Heather J. Zar, Gustavo Wissmann, Ada R.S. Diehl, João C. Prolla, Maria Teresa Jiménez de Anta, José M. Gatell, Paul E. Wilson, Steven R. Meshnick

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79 Citations (Scopus)

Abstract

A polymerase chain reaction (PCR)-based test for Pneumocystis jiroveci (formerly Pneumocystis carinii f. sp. hominis) might be an alternative to histologic diagnoses of P. jiroveci pneumonia (PCP). However, previously developed nested PCR methods tend to have low specificities (high false-positive rates). In this study, nested and quantitative real-time PCR methods for the amplification of the P. jiroveci DHPS (dihydropteroate synthase) gene were evaluated in a variety of stored clinical samples from Spain, South Africa, and Brazil. The sensitivities of both assays were high, ranging from 62.5% to 100% depending on the type of specimen. In a subset of 71 microscopically confirmed PCP cases and 70 negative cases, the sensitivities and specificities were 94% and 81% for nested PCR and 94% and 96% for real-time PCR, respectively. Real-time PCR has a statistically significantly better specificity than nested PCR (P = .015) and is likely to generate fewer false positives. © 2006 Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)153-160
JournalDiagnostic Microbiology and Infectious Disease
Volume56
Issue number2
DOIs
Publication statusPublished - 1 Oct 2006

Keywords

  • Nested PCR
  • Pneumocystis jiroveci
  • Real-time PCR

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