title: Inter-laboratory comparison of three different real-time PCR assays for the detection of Pneumocystis jiroveci in bronchoalveolar lavage fluid samples.
abstract: Pneumocystis jiroveci pneumonia (PCP) is an opportunistic infection affecting immunocompromised patients. While conventional diagnosis of PCP by microscopy is cumbersome, the use of PCR to diagnose PCP has great potential. Nevertheless, inter-laboratory validation and standardization of PCR assays is lacking. The aim of this study was to evaluate the inter-laboratory agreement of three independently developed real-time PCR assays for the detection of P. jiroveci in bronchoalveolar lavage fluid samples. Therefore, 124 samples were collected in three tertiary care laboratories (Leiden University Medical Center, Maastricht Infection Center and Radboud University Nijmegen Medical Centre) and were tested by both microscopy and real-time PCR. Of 41 samples positive for P. jiroveci by microscopy, 40 were positive in all three PCR assays. The remaining sample was positive in a single assay only. Out of 83 microscopy-negative samples, 69 were negative in all three PCR assays. The other 14 samples were found positive, either in all three assays (n=5), in two (n=2) or in one of the assays (n=7). The data demonstrate high inter-laboratory agreement among real-time PCR assays for the detection of P. jiroveci.
authors.affiliation: Department of Medical Microbiology, Maastricht Infection Center (MINC), University Hospital Maastricht, PO Box 5800, 6202 AZ Maastricht, The Netherlands.
authors.forename: Catharina F M
authors.initials: CFM
authors.lastname: Linssen
authors.affiliation: Department of Medical Microbiology, Maastricht Infection Center (MINC), University Hospital Maastricht, PO Box 5800, 6202 AZ Maastricht, The Netherlands.
authors.forename: Jan A
authors.initials: JA
authors.lastname: Jacobs
authors.affiliation: Department of Medical Microbiology, Radboud University Nijmegen Medical Centre (RUNMC), PO Box 9101, 6500 HB Nijmegen, The Netherlands.
authors.forename: Pieter
authors.initials: P
authors.lastname: Beckers
authors.affiliation: Department of Medical Microbiology, Leiden University Medical Center (LUMC), PO Box 9600, 2300 RC Leiden, The Netherlands.
authors.forename: Kate E
authors.initials: KE
authors.lastname: Templeton
authors.affiliation: Department of Medical Microbiology, Radboud University Nijmegen Medical Centre (RUNMC), PO Box 9101, 6500 HB Nijmegen, The Netherlands.
authors.forename: Judith
authors.initials: J
authors.lastname: Bakkers
authors.affiliation: Department of Medical Microbiology, Leiden University Medical Center (LUMC), PO Box 9600, 2300 RC Leiden, The Netherlands.
authors.forename: Ed J
authors.initials: EJ
authors.lastname: Kuijper
authors.affiliation: Department of Medical Microbiology, Radboud University Nijmegen Medical Centre (RUNMC), PO Box 9101, 6500 HB Nijmegen, The Netherlands.
authors.forename: Willem J G
authors.initials: WJG
authors.lastname: Melchers
authors.affiliation: Department of Respiratory Medicine, University Hospital Maastricht, PO Box 5800, 6202 AZ Maastricht, The Netherlands.
authors.forename: Marjolein
authors.initials: M
authors.lastname: Drent
authors.affiliation: Department of Medical Microbiology, Maastricht Infection Center (MINC), University Hospital Maastricht, PO Box 5800, 6202 AZ Maastricht, The Netherlands.
authors.forename: Cornelis
authors.initials: C
authors.lastname: Vink
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D006801:Humans; D008853:Microscopy; D045363:Pneumocystis carinii / Q000235:genetics / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000592:standards*; D013223:Statistics as Topic