Development and Validation of a Liquid Biopsy Assay for Detection of C797S Mutation in cell-free DNA of NSCLC Patients

Picture of a lab equipment.

Students

2021
College

Faculty

Research Assistant Professor of Medicine

Project Summary

My research focuses on developing and validating a highly sensitive clinical assay for the C797S mutation, a mutation associated with drug-resistance in non-small cell lung cancer. Although there is a C797S assay for research use commercially available, it had not been tested on clinical samples; moreover, special pre-assay amplification (pre-amp) procedures needed to make the assay robust for clinical use had not been devised. Thus, in my project, I will design and test pre-amp procedures for the C797S research assay commercially available through BIO-RAD, and validate the sensitivity and specificity of the assay in the clinical setting. Although the project is not yet finished, I have already made progress in optimizing PCR design to prepare the pre-amp procedure of C797 amplicon for multiplexing with other mutation amplicons, so it can be incorporated into a more comprehensive assay that detects multiple mutations.

As a pre-med student with an interest in oncology, this project offered me a unique opportunity to tackle a real-world medical challenge from a translational perspective, as I converted basic science knowledge in assay design to developing and validating a clinically applicable mutation detection assay for NSCLC. Specifically, this opportunity allowed me to apply my skills in cfDNA extraction, pre-amplification, and ddPCR quantification, techniques that I mastered during my PURM research last year to solving a new problem. Through exercising these skills, I gained a deeper understanding in experiment design and in new assay development. As a prospective biology major, I also practiced using my theoretical knowledge in molecular genetics and the scientific method to propose meaningful hypothesis, generate robust data, and carry out critical analysis, which in turn deepened my understanding of the science academically. Since PCR is one of the most commonly used technique in biological research, the new skills that I acquired during the course of this project could be also easily transferred to other projects and benefit my future research career.