Rapid detection of epidermal growth factor receptor mutations with multiplex PCR and primer extension in lung cancer

Journal of Biomedical Science

Research

Ching-Hsiung Lin¹  Nicholas C Hsu²  Ya-Sian Chang³  Jan-Gowth Chang⁴  Kun-Tu Yeh⁵ 

[1] Department of Chest Medicine, Changhua Christian Hospital, Changhua, Taiwan;Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan;Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan;Department of Veterinary Medicine, National Chung Hsiung University, Taichung, Taiwan;Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan;Institute of Clinical Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan;Center for Excellence in Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan;Department of Pathology, Changhua Christian Hospital, Changhua, Taiwan;

关键词: Epidermal Growth Factor Receptor;    Gefitinib;    Single Nucleotide Polymorphism;    Epidermal Growth Factor Receptor Mutation;    Epidermal Growth Factor Receptor Gene;   

DOI  :  10.1186/1423-0127-17-37

 received in 2010-03-18, accepted in 2010-05-12,  发布年份 2010

来源: Springer

【 摘 要 】

Epidermal growth factor receptor (EGFR) kinase domain mutations hyperactivate the kinase and confer kinase addiction of the non-small-cell lung cancer (NSCLC) tumor cells. Almost all of these mutations are located within exons 18-21. The -216 single nucleotide polymorphism in the promoter region is associated with increased EGFR production. We present a method for detecting these common mutations in 81 cases of NSCLC. The protocol is based on the multiplex amplification of promoter region and exons 18-21 of the EGFR genes in a single tube, followed by primer extension of the PCR products using various sizes of primers to detect base changes at -216 promoter region and codons 719, 746-750, 790, 858 of the EGFR gene. We compared the results with that from direct sequencing for detecting EGFR mutations in 81 cases of NSCLC. The two methods identified the same 26 mutations, but our method is superior to direct sequencing in terms of the amount of work and time required. We presented a simple and fast method to detect mutations of EGFR genes in NSCLC.

【 授权许可】

Unknown   
© Lin et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.