Anne Bergougnoux1,2, Valeria D'Argenio3,4, Stefanie Sollfrank5, Fanny Verneau1, Antonella Telese3, Irene Postiglione3, Karl J Lackner5, Mireille Claustres2, Giuseppe Castaldo3,4, Heidi Rossman6, Francesco Salvatore7,8, Caroline Raynal9,10. 1. Laboratory of Molecular Genetics, Montpellier University Hospital, Montpellier Cedex 5, France. 2. Rare Genetic Diseases Laboratory, Institut Universitaire de Recherche Clinique, University of Montpellier, Montpellier Cedex 5, France. 3. CEINGE-Biotecnologie Avanzate, Naples, Italy. 4. Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, Naples, Italy. 5. Department of Clinical Chemistry and Laboratory Medicine, Johannes Gutenberg University, Mainz, Germany. 6. Department of Clinical Chemistry and Laboratory Medicine, Johannes Gutenberg University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany, Phone: +496131177297. 7. CEINGE-Biotecnologie Avanzate, via Gaetano Salvatore 486, 80145 Naples, Italy. 8. Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, via S. Pansini 5, 80131 Naples, Italy, Phone: +390817463648. 9. Laboratory of Molecular Genetics, Montpellier University Hospital, 34093 Montpellier Cedex 5, France. 10. Rare Genetic Diseases Laboratory, Institut Universitaire de Recherche Clinique, EA 7402, University of Montpellier, 34000 Montpellier Cedex 5, France, Phone: +33(0)411759879.
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.
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.
Authors: Manuela Sterrantino; Andrea Fuso; Silvia Pierandrei; Sabina Maria Bruno; Giancarlo Testino; Giuseppe Cimino; Antonio Angeloni; Marco Lucarelli Journal: Diagnostics (Basel) Date: 2021-01-25