title: Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens.
abstract: 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.
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Flori
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Bahrie
authors.initials: B
authors.lastname: Bellete
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Durand
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Raberin
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Cazorla
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Jamal
authors.initials: J
authors.lastname: Hafid
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Lucht
authors.affiliation: Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.
authors.forename: Roger Tran Manh
authors.initials: RTM
authors.lastname: Sung
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis / Q000382:microbiology; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D002353:Carrier State / Q000175:diagnosis / Q000382:microbiology; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D005188:False Negative Reactions; D005189:False Positive Reactions; D006801:Humans; D016867:Immunocompromised Host; D045363:Pneumocystis carinii / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis* / Q000382:microbiology / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004271:DNA, Fungal