title: A misleading false-negative result of Pneumocystis real-time PCR assay due to a rare punctual mutation: A French multicenter study.
abstract: This article describes a previously unreported mutation at position 210 (C210T) of the mitochondrial large subunit ribosomal RNA (mtLSUrRNA) gene of Pneumocystis jirovecii, which led to a false-negative result of a real-time polymerase chain reaction (PCR) assay. Since the aforementioned real-time PCR assay is widely used in France, a French multicenter study was conducted to estimate the mutation frequency and its potential impact on the routine diagnosis of Pneumocystis pneumonia (PCP). Through analysis of data obtained from eight centers, the mutation frequency was estimated at 0.28%. This low frequency should not call into question the routine use of this PCR assay. Nonetheless, the occurrence of the false-negative PCR result provides arguments for maintaining microscopic techniques combined to PCR assays to achieve PCP diagnosis.
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France solene.legal@univ-brest.fr.|Laboratory of Parasitology and Mycology, Brest University Hospital, Brest, France.
authors.forename: Solène
authors.initials: S
authors.lastname: Le Gal
authors.affiliation: University of Rennes 1, INSERM U1085, Rennes, France.|Laboratory of Parasitology and Mycology, Rennes University Hospital, Rennes, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Robert-Gangneux
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Pépino
authors.affiliation: University of Rennes 1, INSERM U1085, Rennes, France.|Laboratory of Parasitology and Mycology, Rennes University Hospital, Rennes, France.
authors.forename: Sorya
authors.initials: S
authors.lastname: Belaz
authors.affiliation: University of Picardy-Jules Verne, EA 4285 UMR-I 01 INERIS, Amiens, France.|Department of Parasitology and Mycology, Amiens University Hospital, Amiens, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Damiani
authors.affiliation: Laboratory of Molecular Genetics and Histocompatibility, Brest University Hospital, Brest, France.|University of Brest, INSERM 1078, Molecular Genetics and Epidemiological Genetics, SFR 148, Brest, France.
authors.forename: Paul
authors.initials: P
authors.lastname: Guéguen
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Pitous
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France.
authors.forename: Michèle
authors.initials: M
authors.lastname: Virmaux
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France.
authors.forename: Eloise
authors.initials: E
authors.lastname: Lissillour
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Pougnet
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France.
authors.forename: Thibaud
authors.initials: T
authors.lastname: Guillaud-Saumur
authors.affiliation: Parasitology and Mycology laboratory, Reims University Hospital, Reims, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Toubas
authors.affiliation: Parasitology and Mycology laboratory, Dijon University Hospital, Dijon, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Valot
authors.affiliation: Parasitology and Mycology laboratory, Saint Antoine Hospital, Assistance Publique-Hôpitaux de Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Hennequin
authors.affiliation: Parasitology and Mycology laboratory, Nantes University Hospital, Nantes, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Morio
authors.affiliation: Parasitology and Mycology laboratory, Nice University Hospital, Nice, France.
authors.forename: Lilia
authors.initials: L
authors.lastname: Hasseine
authors.affiliation: University of Angers, GEIHP EA 3142, Angers, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Bouchara
authors.affiliation: University of Picardy-Jules Verne, EA 4285 UMR-I 01 INERIS, Amiens, France.|Department of Parasitology and Mycology, Amiens University Hospital, Amiens, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Totet
authors.affiliation: University of Brest, GEIHP EA 3142, Brest, France gilles.nevez@chu-brest.fr.|Laboratory of Parasitology and Mycology, Brest University Hospital, Brest, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Nevez
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005188:False Negative Reactions; D005260:Female; D005602:France; D005787:Gene Frequency; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D045363:Pneumocystis carinii / Q000235:genetics / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis* / Q000382:microbiology; D017354:Point Mutation; D012335:RNA, Ribosomal / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D055815:Young Adult
chemical_list: D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D004275:DNA, Ribosomal; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S
keywords: PCP; Pneumocystis jirovecii; Real time-PCR; TaqMan® MGB probe; mutation