Activating mutations within the tyrosine kinase domain of epidermal growth factor receptor (EGFR) are frequently found in non-small-cell lung cancer (NSCLC) patients and are associated with the patients’ response to EGFR inhibitors. EGFR exon 19 deletion is one of the most common NSCLC-associated EGFR mutations. Taking advantages of a novel property of lambda exonuclease observed in our lab, we have developed a fluorophore-modified DNA probe to form different terminal structures with mutant-type and wild-type strands. Lambda exonuclease can sensitively distinguish between the two structures, thus hydrolyzing the probe at different rates. An integral procedure is established to detect EGFR exon 19 deletion in circulating cell-free DNA. This new method can detect mutant DNA at an abundance as low as 0.01% within 4 hours without the requirement of expensive equipments, which is superior over current clinical methods such as next generation sequencing and digital PCR. The method has great potential to be put into clinical practice.

DNA terminal structure-mediated enzymatic reaction for ultra-sensitive detection of EGFR exon 19 deletion

2019 Fall National Meeting

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