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External quality assessment for molecular detection of Bordetella pertussis in European laboratories
G Muyldermans1, O Soetens, M Antoine
1Department of Microbiology, Academisch Ziekenhuis Vrije Universiteit Brussel, Laarbeeklaan 101, 1090 Brussels, Belgium. labomicro@az.vub.ac.be
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.
Insights
External quality assessment for Bordetella pertussis PCR revealed significant variability. Choosing the correct target gene, like pertactin, is crucial for accurate species detection and avoiding false positives in pertussis diagnostics.
Area of Science:
- Clinical microbiology
- Molecular diagnostics
- Infectious disease surveillance
Background:
- Polymerase chain reaction (PCR) is standard for Bordetella pertussis detection.
- No established external quality assessment (EQA) programs exist for pertussis PCR.
- Ensuring accuracy and specificity in pertussis diagnostics is critical.
Purpose of the Study:
- To evaluate the performance of European laboratories in Bordetella pertussis PCR detection.
- To assess the impact of different PCR target genes on species specificity.
- To identify areas for improvement in pertussis diagnostic assays.
Main Methods:
- Two external quality assessment proficiency panels were distributed to European laboratories.
- Panels included dilutions of Bordetella pertussis clinical and reference strains, other Bordetella species, and negative controls.
- Laboratory results were analyzed for accuracy, sensitivity, specificity, and limits of detection.
Main Results:
- No false positives were reported in the first panel, but limits of detection varied widely.
- The second panel revealed one laboratory error and cross-reactivity issues with IS481-based assays.
- Assays targeting the pertactin gene demonstrated 100% specificity for Bordetella pertussis.
Conclusions:
- The choice of PCR target gene significantly impacts the species specificity of Bordetella pertussis detection.
- IS481-based assays show poor specificity, cross-reacting with Bordetella holmesii and Bordetella bronchiseptica.
- Standardized EQA programs and careful target gene selection are essential for reliable pertussis PCR diagnostics.
