title: Proficiency program for real-time PCR diagnosis of Bordetella pertussis infections in French hospital laboratories and at the French National Reference Center for Whooping Cough and other Bordetelloses.
abstract: With the support of a ministerial program for innovative and expensive technologies, dedicated to the economic evaluation of laboratory diagnosis of pertussis by real-time PCR, external quality assessment for real-time IS481 PCR was carried out. Coordinated by the National Centre of Reference of Pertussis and other Bordetelloses (NCR), this study aimed to harmonize and to assess the performances of eight participating microbiology hospital laboratories throughout the French territory. Between January 2006 and February 2007, 10 proficiency panels were sent by the NCR (ascending proficiency program), representing a total of 49 samples and including eight panels to analyze and evaluate the global sensitivity and specificity of real-time PCR, one to assess the limit of detection, and one to evaluate nucleic acid extraction methods. As part of the descending proficiency program, extracted DNA from clinical samples was sent by the eight participating laboratories in different panels and analyzed by the NCR. In the ascending proficiency analysis, the sensitivity and specificity of the real-time PCR methods were 92.2% and 94.3%, respectively. The limit of detection of the different methods ranged between 0.1 and 1 fg/microl (0.2 to 2 CFU/microl). The nucleic acid extraction methods showed similar performances. During the descending proficiency analysis, performed with 126 samples, the result of the NCR for 15 samples (11.9%) was discordant with the result obtained by the source laboratory. Despite several initial differences, harmonization was easy and performances were homogeneous. However, the risk of false-positive results remains quite high, and we strongly recommend establishment of uniform quality control procedures performed regularly.
authors.affiliation: Institut Pasteur, Centre National de Référence (CNR) de la Coqueluche et autres Bordetelloses, Paris, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Caro
authors.forename: Nicole
authors.initials: N
authors.lastname: Guiso
authors.forename: Corinne
authors.initials: C
authors.lastname: Alberti
authors.forename: Sandrine
authors.initials: S
authors.lastname: Liguori
authors.forename: Christophe
authors.initials: C
authors.lastname: Burucoa
authors.forename: Gérard
authors.initials: G
authors.lastname: Couetdic
authors.forename: Florence
authors.initials: F
authors.lastname: Doucet-Populaire
authors.forename: Agnès
authors.initials: A
authors.lastname: Ferroni
authors.forename: Sophie
authors.initials: S
authors.lastname: Papin-Gibaud
authors.forename: Florence
authors.initials: F
authors.lastname: Grattard
authors.forename: Hélène
authors.initials: H
authors.lastname: Réglier-Poupet
authors.forename: Josette
authors.initials: J
authors.lastname: Raymond
authors.forename: Catherine
authors.initials: C
authors.lastname: Soler
authors.forename: Sylvie
authors.initials: S
authors.lastname: Bouchet
authors.forename: Sandrine
authors.initials: S
authors.lastname: Charreau
authors.forename: Brigitte
authors.initials: B
authors.lastname: Couzon
authors.forename: Isabelle
authors.initials: I
authors.lastname: Leymarie
authors.forename: Nicole
authors.initials: N
authors.lastname: Tavares
authors.forename: Mathilde
authors.initials: M
authors.lastname: Choux
authors.forename: Edouard
authors.initials: E
authors.lastname: Bingen
authors.forename: Stéphane
authors.initials: S
authors.lastname: Bonacorsi
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D001886:Bordetella pertussis / Q000235:genetics / Q000302:isolation & purification*; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D005602:France; D006302:Health Services Research; D006801:Humans; D007223:Infant; D007755:Laboratories, Hospital; D016133:Polymerase Chain Reaction / Q000379:methods*; D011785:Quality Assurance, Health Care / Q000379:methods*; D012680:Sensitivity and Specificity; D014917:Whooping Cough / Q000175:diagnosis*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial
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