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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제30권 제5호
- 발행연도
- 2020.1
- 수록면
- 662 - 667 (6page)
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초록· 키워드
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Epidermal growth factor receptor (EGFR) mutations are not only genetic markers for diagnosis but also biomarkers of clinical-response against tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Among the EGFR mutations, the in-frame deletion mutation in EGFR exon 19 kinase domain (EGFR exon 19-del) is the most frequent mutation, accounting for about 45% of EGFR mutations in NSCLCs. Development of sensitive method for detecting the EGFR mutation is highly required to make a better screening for drug-response in the treatment of NSCLC patients. Here, we developed a fluorometric tandem gene amplification assay for sensitive detection of lowabundance EGFR exon 19-del mutant genomic DNA. The method consists of pre-amplification with PCR, thermal cycling of ligation by Taq ligase, and subsequent rolling circle amplification (RCA). PCRamplified DNA from genomic DNA samples was used as splint DNA to conjugate both ends of linear padlock DNA, generating circular padlock DNA template for RCA. Long stretches of ssDNA harboring multiple copies of G-quadruplex structure was generated in RCA and detected by thioflavin T (ThT) fluorescence, which is specifically intercalated into the G-quadruplex, emitting strong fluorescence. Sensitivity of tandem gene amplification assay for detection of the EGFR exon 19-del from gDNA was as low as 3.6 pg, and mutant gDNA present in the pooled normal plasma was readily detected as low as 1% fraction. Hence, fluorometric detection of low-abundance EGFR exon 19 deletion mutation using tandem gene amplification may be applicable to clinical diagnosis of NSCLC patients with appropriate TKI treatment.
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참고문헌 (18)
참고문헌 신청
Siegel RL / 2017 / Cancer Statistics, 2017 / CA Cancer J. Clin 67:7~30
Sharma SV / 2007 / Epidermal growth factor receptor mutations in lung cancer / Nat. Rev. Cancer 7:169~181
Herbst RS / 2018 / The biology and management of non-small cell lung cancer / Nature 553:446~454
Lynch TJ / 2004 / Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib / New Engl. J. Med 350:2129~2139
Tokudome N / 2020 / Differential significance of molecular subtypes which were classified into EGFR exon 19 deletion on the first line afatinib monotherapy / BMC Cancer 20:103
Sharma SV / 2007 / Epidermal growth factor receptor mutations in lung cancer / Nat. Rev. Cancer 7:169~181
Herbst RS / 2018 / The biology and management of non-small cell lung cancer / Nature 553:446~454
Lynch TJ / 2004 / Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib / New Engl. J. Med 350:2129~2139
Tokudome N / 2020 / Differential significance of molecular subtypes which were classified into EGFR exon 19 deletion on the first line afatinib monotherapy / BMC Cancer 20:103
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