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Open AccessCommunication

A Novel Technique to Detect EGFR Mutations in Lung Cancer

by Yuanbin Liu 1,2,†, Ting Lei 3,†, Zhiyu Liu 4,†, Yanbin Kuang 1, Jianxin Lyu 2,* and Qi Wang 1,*
1
Department of Respiratory Medicine, the Second Hospital, Dalian Medical University, Dalian 116023, China
2
Key Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou Medical University, Wenzhou 325035, China
3
Department of Thoracic Surgery, the Second Hospital of Dalian Medical University, Dalian 116023, China
4
Department of Urinary Surgery, the Second Hospital, Dalian Medical University, Dalian 116023, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: William Chi-shing Cho
Int. J. Mol. Sci. 2016, 17(5), 792; https://doi.org/10.3390/ijms17050792
Received: 30 March 2016 / Revised: 17 May 2016 / Accepted: 18 May 2016 / Published: 23 May 2016
Epidermal growth factor receptor (EGFR) gene mutations occur in multiple human cancers; therefore, the detection of EGFR mutations could lead to early cancer diagnosis. This study describes a novel EGFR mutation detection technique. Compared to direct DNA sequencing detection methods, this method is based on allele-specific amplification (ASA), recombinase polymerase amplification (RPA), peptide nucleic acid (PNA), and SYBR Green I (SYBR), referred to as the AS-RPA-PNA-SYBR (ARPS) system. The principle of this technique is based on three continuous steps: ASA or ASA combined with PNA to prevent non-target sequence amplification (even single nucleotide polymorphisms, SNPs), the rapid amplification advantage of RPA, and appropriate SYBR Green I detection (the samples harboring EGFR mutations show a green signal). Using this method, the EGFR 19Del(2) mutation was detected in 5 min, while the EGFR L858R mutation was detected in 10 min. In this study, the detection of EGFR mutations in clinical samples using the ARPS system was compatible with that determined by polymerase chain reaction (PCR) and DNA sequencing methods. Thus, this newly developed methodology that uses the ARPS system with appropriate primer sets is a rapid, reliable, and practical way to assess EGFR mutations in clinical samples. View Full-Text
Keywords: RPA; ASA; SYBR; EGFR mutation; novel methodology; point-of-care test (POCT) RPA; ASA; SYBR; EGFR mutation; novel methodology; point-of-care test (POCT)
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MDPI and ACS Style

Liu, Y.; Lei, T.; Liu, Z.; Kuang, Y.; Lyu, J.; Wang, Q. A Novel Technique to Detect EGFR Mutations in Lung Cancer. Int. J. Mol. Sci. 2016, 17, 792.

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