Objectives: SCN5A mutations lead to a wide spectrum of cardiovascular disorders. Due to large cohorts to investigate and the large gene size, mutational screening must be performed using an extremely sensitive and specific scanning method.
Design and methods: High Resolution Melting (HRM) analysis was developed for SCN5A mutation detection using control DNAs and DNAs carrying previously identified gene variants. A cohort of 40 patients was further screened. To evaluate HRM sensitivity, this cohort was also screened using an optimized DHPLC methodology.
Results: All gene variants detected by DHPLC were also readily identified as abnormal by HRM analysis. Mutations were identified for 5 patients. Complete molecular SCN5A investigation was completed two times faster and cheaper than using DHPLC strategy.
Conclusions: HRM analysis represents an inexpensive, highly sensitive and high-throughput method to allow identification of SCN5A gene variants. Identification of more SCN5A mutations could provide new insights into the pathophysiology of SCN5A-linked diseases syndromes.