Validation of high-resolution DNA melting analysis for mutation scanning of the LMNA gene

Objectives: LMNA mutations lead to a wide spectrum of disorders now called laminopathies. Due to large cohorts to investigate, mutational screening must be performed using an extremely sensitive and specific scanning method. Design and methods: High Resolution Melting (HRM) analysis was developed for LMNA mutation detection. A cohort of 64 patients with dilated cardiomyopathy was prospectively screened using both HRM and DHPLC methodologies. Results: All gene variants detected by DHPLC or by direct sequencing were also readily identified as abnormal by HRM analysis. Mutations were identified in 7 patients (similar to 11%). Complete molecular LMNA 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 identify LMNA genetic variants. The discovery of novel LMNA mutations will provide new insights into the pathophysiology of dilated cardiomyopathy and in all other laminopathies. (C) 2009 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.