Enhanced Ratio of Signals Enables Digital Mutation Scanning for Rare Allele Detection

The use of droplet digital PCR (ddPCR) for low-level DNA mutation detection in cancer, prenatal diagnosis, and infectious diseases is growing rapidly. However, although ddPCR has been implemented successfully for detection of rare mutations at pre-determined positions, no ddPCR adaptation for mutation scanning exists. Yet, frequently, clinically relevant mutations reside on multiple sequence positions in tumor suppressor genes or complex hotspot mutations in oncogenes. Here, we describe a combination of coamplification at lower denaturation temperature PCR (COLD-PCR) with ddPCR that enables digital mutation scanning within approximately 50-bp sections of a target amplicon.

Source: Journal of Molecular Diagnostics - March 13, 2015 Category: Pathology Authors: Elena Castellanos-Rizaldos, Cloud Paweletz, Chen Song, Geoffrey R. Oxnard, Harvey Mamon, Pasi A. Jänne, G. Mike Makrigiorgos Tags: Regular Article Source Type: research