1. Monitoring of Response and Resistance in Plasma of EGFR-Mutant Lung Cancer Using Droplet Digital PCR
- Author
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Nora Feeney, Geoffrey R. Oxnard, Allison O'Connell, Yanan Kuang, Cloud P. Paweletz, Adrian G. Sacher, and Ryan S. Alden
- Subjects
0301 basic medicine ,Mutant ,Genetic analysis ,Molecular biology ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,chemistry ,Cell-free fetal DNA ,030220 oncology & carcinogenesis ,Genotype ,Digital polymerase chain reaction ,Liquid biopsy ,Genotyping ,DNA - Abstract
The identification of oncogenic driver mutations has led to the rapid rise of genotype-directed treatments. However, genetic analysis of tumors remains cumbersome and a morbid experience for patients. Noninvasive assessment of tumor genotype, so-called "liquid biopsy," such as plasma genotyping represents a potentially transformative tool. Here we describe a genotyping protocol of cell-free plasma DNA (cfDNA) using Droplet Digital™ PCR (ddPCR™). ddPCR emulsifies DNA into ~20,000 droplets in which PCR is performed to endpoint in each droplet for both mutant and wild-type DNA. Droplets are run through a modified flow cytometer where mutant and wild-type DNA emit different colored signals. The count of these signals upon Poisson distribution analysis allows sensitive quantification of allelic prevalence.
- Published
- 2018
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