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

Authors
 Suki Kang  ;  Baek Gil Kim  ;  Hyun Ho Han  ;  Joo Hyun Lee  ;  Ji Eun Kim  ;  Hyo Sup Shim  ;  Nam Hoon Cho 
Citation
 Oncotarget, Vol.6(15) : 13742-13749, 2015 
Journal Title
 Oncotarget 
ISSN
 1949-2553 
Issue Date
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.
Files in This Item:
T201502234.pdf Download
DOI
10.18632/oncotarget.3807
Appears in Collections:
1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Pathology (병리학교실) > 1. Journal Papers
Yonsei Authors
Kang, Suki(강숙희) ORCID logo https://orcid.org/0000-0002-9957-3479
Kim, Baek Gil(김백길) ORCID logo https://orcid.org/0000-0001-6270-1433
Kim, Ji Eun(김지은)
Lee, Joo Hyun(이주현)
Cho, Nam Hoon(조남훈) ORCID logo https://orcid.org/0000-0002-0045-6441
Han, Hyun Ho(한현호) ORCID logo https://orcid.org/0000-0002-6268-0860
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/140557
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