Sensitive detection of T790M mutations in lung cancer biopsies using a PCR-based lateral flow assay

T790 M point mutations in EGFR exon 20 are regarded as the most common cause of resistance to epidermal growth factor receptor tyrosine kinase inhibitor treatment. In this study, a PCR-based lateral flow assay (PCR-LFA) was developed to detect T790 M mutations in human genomic DNA. Detection sensitivity was determined using DNA at different mutant to wild-type ratios. The limit of detection of mutant alleles was 15 copies per reaction. The sensitivity of detection of these mutations in 40 formalin-fixed paraffin-embedded tissue biopsies from non-small cell lung cancer patients was analyzed using PCR-LFA and amplification refractory mutation system (ARMS) PCR. Our assay provided the same information as ARMS PCR for 95% (38/40) of the samples. T790 M mutations were detected in 15 (37.5%) and 13 samples using our assay and ARMS PCR, respectively. Droplet digital PCR confirmed the presence of T790 M mutations in the two discordant samples. These results indicate that PCR-LFA is more sensitive than ARMS PCR for clinical screening of these mutations.