Abstract

Objective: Hypertrophic cardiomyopathy (HCM) is one of the most common inherited cardiovascular diseases. To date, at least 1,800 mutations in more than 50 different genes have been reported to be associated with this condition, among which mutations in the TPM1 gene have been identified as causing hypertrophic cardiomyopathy. Previously, we identified a variant in the TPM1 gene (c.576G>C) in a family with individuals affected by hypertrophic cardiomyopathy. Therefore, this study initially aimed to develop and optimize a Tetra-primer ARMS PCR protocol to detect the c.576G>C variant in the TPM1 gene associated with hypertrophic cardiomyopathy.

Methods: Specific primer pairs were designed to detect the G>C mutation site in the TPM1 gene by optimizing the annealing temperature, primer concentration, and the number of PCR cycles in the Tetra-primer ARMS PCR. The protocol was validated using DNA samples from 9 volunteers in the same family, including 3 patients with hypertrophic cardiomyopathy and 6 unaffected members who had been previously analyzed by Sanger sequencing.

Results: The Tetra-primer ARMS PCR protocol showed that 3 patients in the family carried the variant in the TPM1 gene, consistent with the results obtained from previous Sanger sequencing.

Conclusion: This study developed and initially optimized a Tetra-primer ARMS PCR protocol for detecting the c.576G>C variant in the TPM1 gene. The results were consistent with the Sanger sequencing method; however, further studies with larger sample sizes are needed to fully evaluate and confirm the clinical applicability of this protocol.