Multicenter NGS Validation of CE-IVD CFTR Gene Scanning Kit Achieves Over 99% Accuracy
Three European cystic fibrosis reference laboratories validated the Multiplicom CFTR MASTR Dx kit on the Illumina MiSeq platform using 164 previously characterised samples. Analytical sensitivity, specificity, and accuracy all exceeded 99% across two independent bioinformatics pipelines. The single-workflow approach to detecting multiple CFTR mutation types simplifies routine diagnostics and supports the CE-IVD certification pathway now required under European regulations.
The original study
Multicenter validation study for the certification of a CFTR gene scanning method using next generation sequencing technology.
- Authors
- Bergougnoux A, D'Argenio V, Sollfrank S, Verneau F, Telese A, Postiglione I, et al.
- Journal
- Clinical chemistry and laboratory medicine
- Type
- Journal Article, Multicenter Study, Validation Study
- PMID
- 29427548
Original abstract
BACKGROUND: Many European laboratories offer molecular genetic analysis of the CFTR gene using a wide range of methods to identify mutations causative of cystic fibrosis (CF) and CFTR-related disorders (CFTR-RDs). Next-generation sequencing (NGS) strategies are widely used in diagnostic practice, and CE marking is now required for most in vitro diagnostic (IVD) tests in Europe. The aim of this multicenter study, which involved three European laboratories specialized in CF molecular analysis, was to evaluate the performance of Multiplicom's CFTR MASTR Dx kit to obtain CE-IVD certification. METHODS: A total of 164 samples, previously analyzed with well-established "reference" methods for the molecular diagnosis of the CFTR gene, were selected and re-sequenced using the Illumina MiSeq benchtop NGS platform. Sequencing data were analyzed using two different bioinformatic pipelines. Annotated variants were then compared to the previously obtained reference data. RESULTS AND CONCLUSIONS: The analytical sensitivity, specificity and accuracy rates of the Multiplicom CFTR MASTR assay exceeded 99%. Because different types of CFTR mutations can be detected in a single workflow, the CFTR MASTR assay simplifies the overall process and is consequently well suited for routine diagnostics.