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

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 stu...

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Autores principales: Horiuchi, Sho, Saito, Yuichi, Matsui, Atsuka, Takahashi, Nobumasa, Ikeya, Tomohiko, Hoshi, Eishin, Shimizu, Yoshihiko, Yasuda, Masanori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7010225/
https://www.ncbi.nlm.nih.gov/pubmed/32124949
http://dx.doi.org/10.3892/ijo.2020.4961
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author Horiuchi, Sho
Saito, Yuichi
Matsui, Atsuka
Takahashi, Nobumasa
Ikeya, Tomohiko
Hoshi, Eishin
Shimizu, Yoshihiko
Yasuda, Masanori
author_facet Horiuchi, Sho
Saito, Yuichi
Matsui, Atsuka
Takahashi, Nobumasa
Ikeya, Tomohiko
Hoshi, Eishin
Shimizu, Yoshihiko
Yasuda, Masanori
author_sort Horiuchi, Sho
collection PubMed
description 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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spelling pubmed-70102252020-02-14 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 Int J Oncol Articles 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. D.A. Spandidos 2020-01-14 /pmc/articles/PMC7010225/ /pubmed/32124949 http://dx.doi.org/10.3892/ijo.2020.4961 Text en Copyright: © Horiuchi et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Horiuchi, Sho
Saito, Yuichi
Matsui, Atsuka
Takahashi, Nobumasa
Ikeya, Tomohiko
Hoshi, Eishin
Shimizu, Yoshihiko
Yasuda, Masanori
A novel loop-mediated isothermal amplification method for efficient and robust detection of EGFR mutations
title A novel loop-mediated isothermal amplification method for efficient and robust detection of EGFR mutations
title_full A novel loop-mediated isothermal amplification method for efficient and robust detection of EGFR mutations
title_fullStr A novel loop-mediated isothermal amplification method for efficient and robust detection of EGFR mutations
title_full_unstemmed A novel loop-mediated isothermal amplification method for efficient and robust detection of EGFR mutations
title_short A novel loop-mediated isothermal amplification method for efficient and robust detection of EGFR mutations
title_sort novel loop-mediated isothermal amplification method for efficient and robust detection of egfr mutations
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7010225/
https://www.ncbi.nlm.nih.gov/pubmed/32124949
http://dx.doi.org/10.3892/ijo.2020.4961
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