Fast Assays to Detect Interruptions in CTG.CAG Repeat Expansions

Different interrupted repeat expansions have been found in several trinucleotide repeat (TNR) diseases such as fragile X syndrome (FXS), spinocerebellar ataxias (SCAs), and myotonic dystrophies (DMs). Their origins and roles remain poorly understood, especially in myotonic dystrophy type 1 (DM1). We present here the triplet repeat primed polymerase chain reaction (TP-PCR) and restriction enzyme-digested PCR to detect and identify interrupted triplet repeat alleles in DM1. TP-PCR consists of a PCR amplification using a fluoresceinated (FAM) primer flanking the repeat region and a primer pair in CTG.CAG repeats. A detailed analysis of interrupted triplet repeat tracts is essential to fully understand the role of interruptions in the pathogenesis and molecular mechanisms observed in TNR diseases.