title: External quality assessment for molecular detection of Bordetella pertussis in European laboratories.
abstract: Although the PCR for the detection of Bordetella pertussis is routinely performed in diagnostic laboratories, no quality assessment program has so far been described. We report on the results obtained with two external quality assessment proficiency panels sent to European laboratories. The first proficiency panel contained a series of dilutions of three previously characterized B. pertussis clinical isolates and two negative controls. No false-positive results were reported by six laboratories providing seven data sets. The reported limits of detection of the three B. pertussis strains varied between 4 and 4,000, 9 and 9,000, and 3 and 30,000 CFU/ml, respectively. The second proficiency panel, composed of a series of dilutions of reference strains of B. pertussis, B. holmesii, B. hinzii, and B. bronchiseptica, as well as negative controls, was sent to nine laboratories. One laboratory reported a negative result for a sample and reported a B. parapertussis-positive sample to be positive for B. pertussis. By using the B. pertussis-specific target gene pertactin, one laboratory detected B. pertussis with 100% specificity. All other laboratories, which used IS481-based assays, reported positive results for the samples containing B. holmesii and B. bronchiseptica, species that have occasionally been recovered from human respiratory samples. These data show that the choice of the target gene is particularly critical for the species specificity of B. pertussis PCR assays.
authors.affiliation: Department of Microbiology, Academisch Ziekenhuis Vrije Universiteit Brussel, Laarbeeklaan 101, 1090 Brussels, Belgium. labomicro@az.vub.ac.be
authors.forename: G
authors.initials: G
authors.lastname: Muyldermans
authors.forename: O
authors.initials: O
authors.lastname: Soetens
authors.forename: M
authors.initials: M
authors.lastname: Antoine
authors.forename: S
authors.initials: S
authors.lastname: Bruisten
authors.forename: B
authors.initials: B
authors.lastname: Vincart
authors.forename: F
authors.initials: F
authors.lastname: Doucet-Populaire
authors.forename: N K
authors.initials: NK
authors.lastname: Fry
authors.forename: P
authors.initials: P
authors.lastname: Olcén
authors.forename: J M
authors.initials: JM
authors.lastname: Scheftel
authors.forename: J M
authors.initials: JM
authors.lastname: Senterre
authors.forename: A
authors.initials: A
authors.lastname: van der Zee
authors.forename: M
authors.initials: M
authors.lastname: Riffelmann
authors.forename: D
authors.initials: D
authors.lastname: Piérard
authors.forename: S
authors.initials: S
authors.lastname: Lauwers
mesh_terms: D001886:Bordetella pertussis / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis*; D005060:Europe; D005189:False Positive Reactions; D006801:Humans; D007753:Laboratories; D016133:Polymerase Chain Reaction; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014917:Whooping Cough / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial