High-Throughput Diagnostic Assay for a Highly Prevalent Cardiomyopathy-Associated MYBPC3 Variant

Area of Science:

• Genetics
• Cardiovascular Disease
• Molecular Diagnostics

Background:

• A 25-basepair deletion in the MYBPC3 gene is frequent in South Asians, increasing cardiomyopathy risk.
• This variant affects millions globally, necessitating efficient screening and risk stratification.
• Early detection is crucial for managing cardiomyopathy risk in high-prevalence populations.

Purpose of the Study:

• To develop and validate a rapid, cost-effective assay for detecting the MYBPC3 25-basepair deletion variant.
• To enable accurate genetic screening for cardiomyopathy risk in South Asian populations.
• To facilitate early identification of carriers for improved clinical management.

Main Methods:

• Development of an RNaseH quantitative PCR (qPCR) assay with specific nucleotide primers.
• Design of primers to differentiate between wild-type MYBPC3 alleles and the deletion variant.
• Validation of the assay using human genomic DNA from blood and saliva samples with blinded operators.

Main Results:

• The RNaseH qPCR assay accurately identified the MYBPC3 25-basepair deletion variant in DNA from blood and saliva.
• Three blinded operators achieved high accuracy in genotype determination using the developed assay.
• The assay is robust, verifiable, and automatable for large-scale screening.

Conclusions:

• A simple, validated RNaseH qPCR assay can rapidly detect the prevalent MYBPC3 25-basepair deletion.
• This assay facilitates prescreening of individuals at high risk for cardiomyopathy.
• The test supports clinical identification of MYBPC3 variant carriers in epidemiological studies and clinical settings.