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제117회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Fluorometric detection of EGFR exon 19 deletion causing lung cancer with fluorescent DNA and graphene oxide

2016년 2월 25일 14시 16분 07초
BIO.P-311 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
4월 21일 (목요일) 11:00~12:30
저자 및
김동민, 김동은*
건국대학교 생명공학과, Korea
Mutations in epidermal growth factor receptor (EGFR) have been known as biomarker that causes non-small cell lung cancer. Especially, about 50 % of NSCLC patients possess deletion of several sequences in exon 19 of EGFR gene. As less invasive method for detection of EGFR exon 19 deletion mutation is required, we developed a simple PCR-based detection of EGFR exon 19 deletion by using quencher-free fluorescent probe DNA and graphene oxide (GO). In the presence of the exon 19 deletion mutation, the fluorophore-labeled DNA probe was designed to be fully complementary the mutant sequences. The fully annealed DNA probe was then degraded by the 5′ to 3′ exonuclease activity of Taq DNA polymerase during PCR, releasing the fluorophore from the probe DNA. In contrast, wild type of exon 19 gene would allow the probe DNA to be annealed partially to the template DNA due to absence of the deletion. The partially annealed probe DNA would be digested by Taq polymerase, releasing a fragmental probe DNA. When GO was added to each reaction solution, it produced different fluorescence signals; enhanced fluorescence was observed due to the released fluorophore from the probe DNA that was not adsorbed onto GO, whereas fluorescence was quenched with fragmented single stranded probe DNA that was easily adsorbed onto GO. We demonstrated that the fluorescence signal caused by the exon 19 deletion mutation gene was quantitatively correlated with amount of the mutant gene. Thus, we believe that the GO-based fluorometric assay can be applicable for diagnostic detection of the exon 19 deletion in EGFR gene for early detection of NSCLC.