High throughput screening for expanded CTG repeats in myotonic dystrophy type 1 using melt curve analysis.

ABSTRACT:

Background

Myotonic dystrophy type 1 (DM1) is caused by CTG repeat expansions in the DMPK gene and is the most common form of muscular dystrophy. Patients can have long delays from onset to diagnosis, since clinical signs and symptoms are often nonspecific and overlapping with other disorders. Clinical genetic testing by Southern blot or triplet-primed PCR (TP-PCR) is technically challenging and cost prohibitive for population surveys.

Methods

Here, we present a high throughput, low-cost screening tool for CTG repeat expansions using TP-PCR followed by high resolution melt curve analysis with saturating concentrations of SYBR GreenER dye.

Results

We determined that multimodal melt profiles from the TP-PCR assay are a proxy for amplicon length stoichiometry. In a scree

SUBMITTER: Butterfield RJ 

PROVIDER: S-EPMC8123750 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature