A novel loop‑mediated isothermal amplification method for efficient and robust detection of EGFR mutations

Horiuchi,Sho; Saito,Yuichi; Matsui,Atsuka; Takahashi,Nobumasa; Ikeya,Tomohiko; Hoshi,Eishin; Shimizu,Yoshihiko; Yasuda,Masanori

doi:10.3892/ijo.2020.4961

A novel loop‑mediated isothermal amplification method for efficient and robust detection of EGFR mutations

Authors:
- Sho Horiuchi
- Yuichi Saito
- Atsuka Matsui
- Nobumasa Takahashi
- Tomohiko Ikeya
- Eishin Hoshi
- Yoshihiko Shimizu
- Masanori Yasuda
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Affiliations: Department of Pathology, Saitama Medical University International Medical Center, Hidaka 350‑1298, Japan, Department of Thoracic Surgery, Saitama Cardiovascular and Respiratory Center, Kumagaya, Saitama 360‑0197, Japan, Fundamental Research Laboratory, Fundamental Technology Research Department, Eiken Chemical Co., Ltd., Otawara, Tochigi 324‑0036, Japan, Department of Surgery, Teikyo University School of Medicine, Tokyo 173‑8605, Japan, Department of Pathology, Saitama Cardiovascular and Respiratory Center, Kumagaya, Saitama 360‑0197, Japan
Published online on: January 14, 2020 https://doi.org/10.3892/ijo.2020.4961
Pages: 743-749
Copyright: © Horiuchi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The activation of somatic mutations conferring sensitivity to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors has been widely used in the development of advanced or metastatic primary lung cancer therapy. Therefore, identification of EGFR mutations is essential. In the present study, a loop‑mediated isothermal amplification (LAMP) method was used to identify EGFR mutations, and its efficiency was compared with the Therascreen quantitative PCR assay. Using LAMP and Therascreen to analyze surgically resected tissue samples from patients with pulmonary adenocarcinoma, EGFR mutations were observed in 32/59 tumor samples (LAMP) and 33/59 tumor samples (Therascreen). Notably, the LAMP assay identified one tumor as wild‑type, which had previously been identified as a deletion mutation in exon 19 via the Therascreen assay (Case X). However, the direct sequencing to confirm the EGFR status of the Case X adhered to the results of the LAMP assay. Further experiments using Case X DNA identified this exon 19 deletion mutation using both methods. In addition, a novel deletion mutation in exon 19 of the EGFR was identified. Overall, the present study shows that the LAMP method may serve as a valuable alternative for the identification oncogene mutations.

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