Development of a high resolution melting method for the detection of genetic variations in hypertrophic cardiomyopathy

详细信息    查看全文

• 作者：Gilles Millat ; Valé ; rie Chanavat ; Hervé ; Cré ; halet ; Robert Rousson
• 关键词：Mutations ; Hypertrophic cardiomyopathy ; High resolution melting ; DHPLC ; Polymorphisms ; Molecular diagnosis
• 刊名：Clinica Chimica Acta
• 出版年：2010
• 出版时间：14 December 2010
• 年：2010
• 卷：411
• 期：23-24
• 页码：1983-1991
• 全文大小：667 K

文摘

lass=""h4"">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.

lass=""h4"">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.

lass=""h4"">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.

lass=""h4"">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.