Development of a high resolution melting method for the detection of genetic variations in hypertrophic cardiomyopathy.
BACKGROUND
Hypertrophic cardiomyopathy (HCM) is the most common genetic cardiac disease affecting 1 in 500 people. Due to large cohorts to investigate, the number of disease-causing genes, the size of the 2 prevalent mutated genes, and the presence of a large spectrum of private mutations, mutational screening must be performed using an extremely sensitive and specific scanning method.


METHODS
High Resolution Melting (HRM) analysis was developed for prevalent HCM-causing genes (MYBPC3, MYH7, TNNT2, and TNNI3) using control DNAs and DNAs carrying previously identified gene variants. A cohort of 34 HCM patients was further blindly 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. Mutational screening of a cohort of 34 HCM cases led to identification of 19 mutated alleles. Complete molecular 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 mutations in the coding sequences of prevalent HCM genes. Identification of more HCM mutations will provide new insights into genotype/phenotype relationships and will allow a better knowledge of the HCM physiopathology.
Millat
Gilles
G
Laboratoire de Cardiogénétique Moléculaire, Centre de Biologie et Pathologie Est, Hospices Civils de Lyon, Lyon, France. gilles.millat@chu-lyon.fr
Chanavat
Valérie
V
Créhalet
Hervé
H
Rousson
Robert
R
D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D005615:Freezing; D014644:Genetic Variation; D006801:Humans; D009691:Nucleic Acid Denaturation; D017422:Sequence Analysis, DNA