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Journal of Clinical Microbiology, October 2009, p. 3197-3203, Vol. 47, No. 10
0095-1137/09/$08.00+0 doi:10.1128/JCM.00817-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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
,
Valérie Caro,1,
Nicole Guiso,1*
Corinne Alberti,2
Sandrine Liguori,3
Christophe Burucoa,4
Gérard Couetdic,5
Florence Doucet-Populaire,6,
Agnès Ferroni,7
Sophie Papin-Gibaud,8
Florence Grattard,9
Hélène Réglier-Poupet,10
Josette Raymond,10
Catherine Soler,9
Sylvie Bouchet,8
Sandrine Charreau,4
Brigitte Couzon,6
Isabelle Leymarie,7
Nicole Tavares,10
Mathilde Choux,5
Edouard Bingen,3 and
Stéphane Bonacorsi3
Institut Pasteur, Centre National de Référence (CNR) de la Coqueluche et autres Bordetelloses, Paris, France,1 Unité d'Epidémiologie Clinique, Hôpital Robert-Debré, APHP, Paris, France,2 Services de Microbiologie, Hôpital Robert-Debré, APHP, Paris, France,3 CHU La Milétrie, Poitiers, France,4 CHU Jean Minjoz, Besançon, France,5 C.H. de Versailles, Le Chesnay, France,6 Hôpital Necker-enfants malades, APHP, Paris, France,7 CHU Nantes, Nantes, France,8 Hôpital Nord, Saint Etienne, France,9 Hôpital Cochin Saint Vincent de Paul, APHP, Paris, France,10
Received 23 April 2009/ Returned for modification 8 June 2009/ Accepted 10 August 2009
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/µl (0.2 to 2 CFU/µl). 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.
* Corresponding author. Mailing address: Institut Pasteur, Unite PTMMH, URA-CNRS 3012, 25 rue du Dr Roux, 75015 Paris, France. Phone: 33 1 45 68 83 34. Fax: 33 1 40 61 35 33. E-mail: nicole.guiso{at}pasteur.fr
Published ahead of print on 19 August 2009.
Supplemental material for this article may be found at http://jcm.asm.org/.
Present address: Institut Pasteur, Genotyping of Pathogens and Public Health, Paris, France.
Present address: Hôpital Antoine Béclère, APHP, Clamart, France.
Journal of Clinical Microbiology, October 2009, p. 3197-3203, Vol. 47, No. 10
0095-1137/09/$08.00+0 doi:10.1128/JCM.00817-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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