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.
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.
Caro
Valérie
V
Institut Pasteur, Centre National de Référence (CNR) de la Coqueluche et autres Bordetelloses, Paris, France.
Guiso
Nicole
N
Alberti
Corinne
C
Liguori
Sandrine
S
Burucoa
Christophe
C
Couetdic
Gérard
G
Doucet-Populaire
Florence
F
Ferroni
Agnès
A
Papin-Gibaud
Sophie
S
Grattard
Florence
F
Réglier-Poupet
Hélène
H
Raymond
Josette
J
Soler
Catherine
C
Bouchet
Sylvie
S
Charreau
Sandrine
S
Couzon
Brigitte
B
Leymarie
Isabelle
I
Tavares
Nicole
N
Choux
Mathilde
M
Bingen
Edouard
E
Bonacorsi
Stéphane
S
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*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial
Arch Pediatr. 2001 Sep;8 Suppl 4:705s-711s
11582916
PLoS One. 2008 Jun 18;3(6):e2437
18560590
Eur J Clin Microbiol Infect Dis. 2008 Feb;27(2):145-8
17965893
J Clin Microbiol. 2000 Jan;38(1):467
10681208
J Clin Microbiol. 2005 Jan;43(1):30-5
15634947
J Med Microbiol. 2006 Aug;55(Pt 8):1043-1051
16849724
J Clin Microbiol. 2003 Sep;41(9):4121-6
12958235
J Clin Microbiol. 2006 Dec;44(12):4577-83
17065269
Infect Control Hosp Epidemiol. 2004 Nov;25(11):995-7
15566036
Pediatr Infect Dis J. 2005 May;24(5 Suppl):S93-7
15876935
J Med Microbiol. 2001 May;50(5):436-440
11339251
J Clin Microbiol. 2008 May;46(5):1672-7
18367565
Nat Genet. 2003 Sep;35(1):32-40
12910271
Infect Control Hosp Epidemiol. 2005 Mar;26(3):288-92
15796282
J Clin Microbiol. 2008 Nov;46(11):3636-8
18768655
Vaccine. 2009 Feb 5;27(6):803-14
19071179
J Clin Microbiol. 2008 Oct;46(10):3228-31
18753352
J Clin Microbiol. 2001 May;39(5):1963-6
11326023
J Clin Microbiol. 2005 Oct;43(10):4925-9
16207944
J Clin Microbiol. 2000 Jun;38(6):2330-3
10834997
Lancet Infect Dis. 2002 Dec;2(12):744-50
12467690