Inter-laboratory comparison of three different real-time PCR assays for the detection of Pneumocystis jiroveci in bronchoalveolar lavage fluid samples

• Published in: Journal of medical microbiology Vol. 55; no. 9; pp. 1229 - 1235
• Main Authors: Linssen, Catharina F. M, Jacobs, Jan A, Beckers, Pieter, Templeton, Kate E, Bakkers, Judith, Kuijper, Ed J, Melchers, Willem J. G, Drent, Marjolein, Vink, Cornelis
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.09.2006; Society for General Microbiology
• Subjects: Biological and medical sciences; Bronchoalveolar Lavage Fluid - microbiology; Fundamental and applied biological sciences. Psychology; Humans; Infectious diseases; Medical sciences; Microbiology; Microscopy; Miscellaneous; Mycology; Pneumocystis carinii - genetics; Pneumocystis carinii - isolation & purification; Pneumocystis jiroveci; Pneumonia, Pneumocystis - diagnosis; Polymerase Chain Reaction - standards; Statistics as Topic; Fungi; Polymerase chain reaction; Microbiology; Bronchoalveolar lavage; Pneumocystis; Fungi Imperfecti; Detection; Real time; Thallophyta
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.46552-0

1 Department of Medical Microbiology, Maastricht Infection Center (MINC), University Hospital Maastricht, PO Box 5800, 6202 AZ Maastricht, The Netherlands 2 Department of Medical Microbiology, Radboud University Nijmegen Medical Centre (RUNMC), PO Box 9101, 6500 HB Nijmegen, The Netherlands 3 Department of Medical Microbiology, Leiden University Medical Center (LUMC), PO Box 9600, 2300 RC Leiden, The Netherlands 4 Department of Respiratory Medicine, University Hospital Maastricht, PO Box 5800, 6202 AZ Maastricht, The Netherlands Correspondence Catharina F. M. Linssen klin{at}lmib.azm.nl Received 3 February 2006 Accepted 15 June 2006 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 . Abbreviations: BAL, bronchoalveolar lavage; DHPS, dihydroperoate synthase; LUMC, Leiden University Medical Center; MINC, Maastricht Infection Center; MSG, major surface glycoprotein; PCP, Pneumocystis jiroveci pneumonia; RUNMC, Radboud University Nijmegen Medical Centre.