Research Article
Paul Griffith, David Sun, S
Abstract
Development of novel monoclonal antibodies, vaccines and oncolytic virus therapies have relied on analysis of biomarkers as potential predictors of success. One well studied biomarker is the CD16/ FcγRIIIa receptor residue 158 F/V. Identifying variants through genotyping of the FcγRIIIa locus is widely practiced and highly varied with commonly used methods including: Sanger sequencing, flow-cytometry, PCR/RFLP, Goldengate (replaced by Infinium) and TaqMAN analysis. While each of these methods have considerable backing in publications related to CD16 FcγRIIIa 158 F/V, the majority present significant short comings in identifying both homozygotes (wild-type and mutant) and heterozygotes in a time and cost-efficient manner. Utilization of droplet-digital PCR with FcγRIIIa-F158V specific probes results in the accurate genotyping using direct recognition of sequence in genomic samples at a lower average cost and faster turnaround. Here we demonstrate the use of ddPCR to accurately identify FcγRIIIa-F158V genotypes with confirmation by Illumina sequencing in 128 patient samples.