Nanofluidic allele-specific digital PCR method for quantifying IDH1 and IDH2 mutation burden in acute myeloid leukemia.
Wiseman, Daniel H Somervaille, Tim C P
Wiseman, Daniel H
Somervaille, Tim C P
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Abstract
Precise quantitation of allelic burden for a pathogenic mutation has diverse clinical and research applications but can be difficult to achieve with conventional qPCR-based techniques, especially at lower mutant allele frequencies. Digital PCR overcomes many of the limitations of qPCR and can be highly quantitative even for single-nucleotide variants, with distinct advantages over next-generation sequencing approaches. Here we describe a method combining the principles of TaqMan(®)-chemistry SNP genotyping with microfluidic digital PCR to generate a highly sensitive, quantitative allele-specific digital PCR assay for the six most common IDH1 and IDH2 mutations encountered in myeloid malignancy. The concept and approach could easily be applied to other suitable SNVs.
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Date
2017
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Article
Citation
Nanofluidic allele-specific digital PCR method for quantifying IDH1 and IDH2 mutation burden in acute myeloid leukemia. 2017, 1633:235-255 Methods Mol. Biol.