Rapid detection of K-, N-, H-RAS, and BRAF hotspot mutations in thyroid cancer using the multiplex primer extension
详细信息 查看全文
- 作者：Ya-Sian Chang ; I-Ling Lin ; Kun-Tu Yeh ; Jan-Gowth Chang
- 关键词：RAS genes ; BRAF gene ; Thyroid cancer ; Multiplex PCR and primer extension
- 刊名：Clinical Biochemistry
- 出版年：2013
- 出版时间：October, 2013
- 年：2013
- 卷：46
- 期：15
- 页码：1572-1577
- 全文大小：502 K
文摘
Objectives
The objective of this study is to develop a multiplex PCR and primer extension to detect K-, N-, H-RAS, and BRAF mutations.Design and methods
DNA samples were isolated from 76 thyroid cancer patients. Multiplex amplification of exons 2 and 3 of three RAS genes and exon 15 of the BRAF gene using three pairs of primers was performed in a single tube. The products were split into three tubes. First, we used nine different-sized N-RAS and BRAF primers to detect base changes in N-RAS and BRAF. The other two tubes used seven separate different-sized K-RAS and H-RAS primers to detect base changes.
Results
We compared these results with direct sequencing. The two methods generated identical results, but our method was superior to direct sequencing in terms of the amount of work and time involved.
Conclusions
We present a rapid method to detect mutations of K-, N-, H-RAS, and BRAF in human cancers.