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

Lin CH, Yeh KT, Chang YS, Hsu NC, Chang JG - J. Biomed. Sci. (2010)

Bottom Line: The -216 single nucleotide polymorphism in the promoter region is associated with increased EGFR production.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.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.

ABSTRACT
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.

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Related in: MedlinePlus

Detection of wild-type and mutant EGFR by primer extension analysis. NSCLC DNA samples of wild-type EGFR and ones containing the following mutations: -216 G/T, 2235-2249 del, 2236-2250 del, 2240-2257 del, and 2573 T>G.
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Figure 1: Detection of wild-type and mutant EGFR by primer extension analysis. NSCLC DNA samples of wild-type EGFR and ones containing the following mutations: -216 G/T, 2235-2249 del, 2236-2250 del, 2240-2257 del, and 2573 T>G.

Mentions: We used multiplex PCR plus primer extension method to detect EGFR -216 promoter region and exons 18-21 mutations in 81 cases of NSCLC (Figure 1). Histologically, there were 26 adenocarcinomas, 6 bronchioloaveolar carcinomas, 33 squamous cell carcinomas, 5 adenosquamous carcinomas, and 11 other types of NSCLCs.


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

Lin CH, Yeh KT, Chang YS, Hsu NC, Chang JG - J. Biomed. Sci. (2010)

Detection of wild-type and mutant EGFR by primer extension analysis. NSCLC DNA samples of wild-type EGFR and ones containing the following mutations: -216 G/T, 2235-2249 del, 2236-2250 del, 2240-2257 del, and 2573 T>G.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2875208&req=5

Figure 1: Detection of wild-type and mutant EGFR by primer extension analysis. NSCLC DNA samples of wild-type EGFR and ones containing the following mutations: -216 G/T, 2235-2249 del, 2236-2250 del, 2240-2257 del, and 2573 T>G.
Mentions: We used multiplex PCR plus primer extension method to detect EGFR -216 promoter region and exons 18-21 mutations in 81 cases of NSCLC (Figure 1). Histologically, there were 26 adenocarcinomas, 6 bronchioloaveolar carcinomas, 33 squamous cell carcinomas, 5 adenosquamous carcinomas, and 11 other types of NSCLCs.

Bottom Line: The -216 single nucleotide polymorphism in the promoter region is associated with increased EGFR production.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.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus