Mass spectrometry based trinucleotide repeat sequence detection using target fragment assay

Abstract

Trinucleotide repeat detection is important for understanding the molecular mechanisms of repeat size mutation and clinical diagnosis. In this study, a novel target fragment assay (TFA) for trinucleotide repeat length detection was developed using magnetic capture and acidic degradation of target polymerase chain reaction amplicons followed by mass spectrometry detection. The developed TFA enables accurate, rapid, and sensitive detection of the trinucleotide repeat lengths and provides a new paradigm for identifying nucleic acid sequence variations. Detection of the CAG repeat length associated with Huntington's disease was successfully demonstrated utilizing the developed TFA.