Pitfalls in genetic testing: the story of missed SCN1A mutations

Background Sanger sequencing, still the standard technique for genetic testing in most diagnostic laboratories and until recently widely used in research, is gradually being complemented by next‐generation sequencing (NGS). No single mutation detection technique is however perfect in identifying all mutations. Therefore, we wondered to what extent inconsistencies between Sanger sequencing and NGS affect the molecular diagnosis of patients. Since mutations in SCN1A, the major gene implicated in epilepsy, are found in the majority of Dravet syndrome (DS) patients, we focused on missed SCN1A mutations. Methods We sent out a survey to 16 genetic centers performing SCN1A testing. Results We collected data on 28 mutations initially missed using Sanger sequencing. All patients were falsely reported as SCN1A mutation‐negative, both due to technical limitations and human errors. Conclusion We illustrate the pitfalls of Sanger sequencing and most importantly provide evidence that SCN1A mutations are an even more frequent cause of DS than already anticipated. We explored to what extent inconsistencies between Sanger sequencing and next‐generation sequencing affect the molecular diagnosis of patients. Hereto we focused on the analysis of mutations in SCN1A, the major gene implicated in Dravet syndrome and epilepsy. We illustrate the pitfalls of genetic screening technologies and most importantly provide evidence that SCN1A mutations are an even more frequent cause of Dravet syndrome than already anticipated.