title: Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples.
abstract: The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.
authors.affiliation: Department of Medical Microbiology, Albert Schweitzer Hospital, Dordrecht, The Netherlands; Department of Pathology, Albert Schweitzer Hospital, Dordrecht, The Netherlands. Electronic address: B.de.Leeuw@RLM-microbiologie.nl.
authors.forename: Bertie H C G M
authors.initials: BH
authors.lastname: de Leeuw
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: W Sebastiaan
authors.initials: WS
authors.lastname: Voskuil
authors.affiliation: Department of Medical Microbiology, Albert Schweitzer Hospital, Dordrecht, The Netherlands.
authors.forename: Boulos
authors.initials: B
authors.lastname: Maraha
authors.affiliation: Department of Medical Microbiology, Maasstad Hospital, Rotterdam, The Netherlands.
authors.forename: Anneke
authors.initials: A
authors.lastname: van der Zee
authors.affiliation: Department of Pathology, Albert Schweitzer Hospital, Dordrecht, The Netherlands.
authors.forename: Pieter J
authors.initials: PJ
authors.lastname: Westenend
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Johannes G
authors.initials: JG
authors.lastname: Kusters
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D016612:Paraffin Embedding; D045363:Pneumocystis carinii / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016707:Tissue Fixation
chemical_list: D004269:DNA, Bacterial; D005557:Formaldehyde
keywords: Formalin-fixed paraffin-embedded; PCR; Pneumocystis jirovecii