Multicenter validation study for the certification of a CFTR gene scanning method using next generation sequencing technology
-
Anne Bergougnoux
,
Francesco Salvatore
and
Caroline Raynal
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.
Acknowledgments
The authors thank Jean Ann Gilder (Scientific Communication srl., Naples, Italy) for writing assistance, and Vittorio Lucignano (CEINGE–Biotecnologie Avanzate), Jessica Varilh and Magali Taulan-Cadars (Laboratoire de Génétique Moléculaire, EA7402, Montpellier) for technical assistance. This study would not have been possible without the expertise of the diagnostics teams of the three laboratories over many years (Dr. Marie des Georges, Jean-Pierre Altieri and Dr. Paola Nardiello).
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: This study was supported by Multiplicom (Niel, Belgium). The three participating centers have signed a partnership agreement with the company until the end of the CE-IVD validation study.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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Supplemental Material:
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2017-0553).
©2018 Walter de Gruyter GmbH, Berlin/Boston
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