title: High-Throughput Diagnostic Assay for a Highly Prevalent Cardiomyopathy-AssociatedMYBPC3Variant.
abstract: A 25-basepair deletion variant ofMYBPC3occurs at high frequency in individuals of South Asian descent and is estimated to affect 55 million people worldwide, carrying an increased likelihood of cardiomyopathy. Since this variant is prevalent and severe in this subpopulation, quick and affordable screening to provide risk-assessment to guide treatment for these patients is critical. An RNaseH qPCR assay was developed to quickly and specifically diagnose the presence of the 25-basepair deletion variant inMYBPC3. RNAseH-blocked nucleotide primers were designed to identify the presence or absence of the wild typeMYBPC3allele or the genomic sequence containing the 25-basepair deletion. Using this assay, three blinded operators were able to accurately determine the genotype from human genomic DNA samples from blood and saliva using a qPCR thermocycler. Furthermore, positive variant subjects were examined by both electrocardiography and echocardiography for the presence of cardiomyopathy. A simple, robust assay was established, verified and validated that can be automated to detect the presence of the highly prevalent 25-basepair deletionMYBPC3variant using both blood and saliva samples. The assay will provide quick and accurate prescreening of individuals at high risk for cardiomyopathies and allow for better clinical identification of 25-basepair deletionMYBPC3carriers in large cohort epidemiological studies.
authors.affiliation: Department of Cell and Molecular Physiology, Loyola University, Chicago, USA; Center for Genetic Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
authors.forename: David Y
authors.initials: DY
authors.lastname: Barefield
authors.affiliation: Department of Cell and Molecular Physiology, Loyola University, Chicago, USA.
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Lynch
authors.affiliation: Department of Cell and Molecular Physiology, Loyola University, Chicago, USA.
authors.forename: Aravindakshan
authors.initials: A
authors.lastname: Jagadeesan
authors.affiliation: Department of Cardiology and Echocardiography and Cardiographics, Loyola University, Chicago, USA.
authors.forename: Thriveni
authors.initials: T
authors.lastname: Sanagala
authors.affiliation: Department of Cell and Molecular Physiology, Loyola University, Chicago, USA.
authors.forename: Sakthivel
authors.initials: S
authors.lastname: Sadayappan
keywords: DNA diagnostic test; Genotype-phenotype; Hypertrophic cardiomyopathy; RNaseH qPCR; South Asian population