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Targeted sequencing with enrichment PCR: a novel diagnostic method for the detection of EGFR mutations.

Title
Targeted sequencing with enrichment PCR: a novel diagnostic method for the detection of EGFR mutations.
Authors
Suki Kang;Baek Gil Kim;Nam Hoon Cho;Hyo Sup Shim;Ji Eun Kim;Joo Hyun Lee;Hyun Ho Han
Issue Date
2015
Journal Title
Oncotarget
ISSN
1949-2553
Citation
Oncotarget, Vol.6(15) : 13742~13749, 2015
Abstract
Epidermal growth factor receptor (EGFR) is an important mediator of tumor cell survival and proliferation. The detection of EGFR mutations can predict prognoses and indicate when treatment with EGFR tyrosine kinase inhibitors should be used. As such, the development of highly sensitive methods for detecting EGFR mutations is important. Targeted next-generation sequencing is an effective method for diagnosing mutations. We compared the abilities of enrichment PCR followed by ultra-deep pyrosequencing (UDP), UDP alone, and PNA-mediated RT-PCR clamping to detect low-frequency EGFR mutations in tumor cell lines and tissue samples. Using enrichment PCR-UDP, we were able to detect the E19del and L858R mutations at minimum frequencies of 0.01% and 0.05%, respectively, in the PC-9 and H197 tumor cell lines. We also confirmed the sensitivity of detecting the E19del mutation by performing a titration analysis in FFPE tumor samples. The lowest mutation frequency detected was 0.0692% in tissue samples. EGFR mutations with frequencies as low as 0.01% were detected using enrichment PCR-UDP, suggesting that this method is a valuable tool for detecting rare mutations, especially in scarce tissue samples or those with small quantities of DNA.
URI

http://ir.ymlib.yonsei.ac.kr/handle/22282913/140557
DOI
10.18632/oncotarget.3807
Appears in Collections:
1. 연구논문 > 1. College of Medicine > Dept. of Life Science
1. 연구논문 > 1. College of Medicine > Dept. of Pathology
Yonsei Authors
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