Chapter title |
Nanofluidic Allele-Specific Digital PCR Method for Quantifying IDH1 and IDH2 Mutation Burden in Acute Myeloid Leukemia
|
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Chapter number | 15 |
Book title |
Acute Myeloid Leukemia
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7142-8_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7140-4, 978-1-4939-7142-8
|
Authors |
Wiseman, Daniel H., Somervaille, Tim C. P., Daniel H. Wiseman, Tim C. P. Somervaille |
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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