Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens

• Published in: Journal of medical microbiology Vol. 53; no. 7; pp. 603 - 607
• Main Authors: Flori, Pierre, Bellete, Bahrie, Durand, Fabrice, Raberin, Helene, Cazorla, Celine, Hafid, Jamal, Lucht, Frederic, Sung, Roger Tran Manh
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.07.2004; Society for General Microbiology
• Subjects: AIDS-Related Opportunistic Infections - diagnosis; AIDS-Related Opportunistic Infections - microbiology; Biological and medical sciences; Bronchoalveolar Lavage Fluid - microbiology; Carrier State - diagnosis; Carrier State - microbiology; DNA, Fungal - analysis; DNA, Fungal - isolation & purification; False Negative Reactions; False Positive Reactions; Fundamental and applied biological sciences. Psychology; Human immunodeficiency virus; Humans; Immunocompromised Host; Infectious diseases; Medical sciences; Microbiology; Miscellaneous; Mycology; Pneumocystis carinii; Pneumocystis carinii - cytology; Pneumocystis carinii - genetics; Pneumocystis carinii - isolation & purification; Pneumocystis jiroveci; Pneumonia, Pneumocystis - diagnosis; Pneumonia, Pneumocystis - microbiology; Pneumonia, Pneumocystis - pathology; Polymerase Chain Reaction - methods; Sensitivity and Specificity; Staining and Labeling - methods; Fungi; Polymerase chain reaction; Lung disease; Pneumonia; Microbiology; Respiratory disease; Bronchoalveolar lavage; Pneumocystis; Fungi Imperfecti; Real time; Thallophyta
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.45528-0

Laboratory of Parasitology and Mycology, Hôpital Nord 1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue 2 , University Hospital of Saint Etienne, 42055 Saint Etienne, France Correspondence Pierre Flori pierre.flori{at}univ-st-etienne.fr Received November 7, 2003 Accepted February 10, 2004 Between January 2002 and July 2003, 173 bronchoalveolar lavage (BAL) specimens from 150 patients (19 HIV-infected and 131 non-HIV-infected patients) were evaluated for identification of Pneumocystis jiroveci (formerly known as Pneumocystis carinii f. sp. hominis ) using staining techniques, conventional PCR ( mtLSUrRNA gene) and real-time PCR ( MSG gene). Test results were compared to Pneumocystis pneumonia (PCP) confirmed by typical clinical findings and response to treatment. Sensitivity and specificity of the techniques were 60 and 100 % for staining (where either one or both techniques were positive), 100 and 87.0 % for conventional PCR and 100 and 84.9 % for real-time PCR, respectively. The use of a concentration of 10 3 copies of DNA per capillary of BAL as a cut-off (determined by real-time PCR) increased specificity from 84.9 to 98.6 % without reducing the sensitivity of the technique. This technique is rapid (<3 h) and therefore of major interest in differentiating between asymptomatic carriage and PCP. A BAL specimen with <10 3 copies per capillary of Pneumocystis -specific DNA is more likely to indicate a chronic carrier state, but in such cases follow-up is required to ensure that the patient is not in the early stage of an active PCP. Abbreviations: BAL, bronchoalveolar lavage; PCP, Pneumocystis pneumonia.