Rapid and Simple Detection of Hot Spot Point Mutations of Epidermal Growth Factor Receptor, BRAF, and NRAS in Cancers Using the Loop-Hybrid Mobility Shift Assay

摘要: A simple and rapid method to detect the epidermal growth factor receptor hot spot mutation L858R in lung adenocarcinoma was developed based on principles similar universal heteroduplex generator technology. single-stranded oligonucleotide with an internal deletion used generate heteroduplexes (loop-hybrids) bearing a loop complementary strand derived from polymerase chain reaction product of normal or mutant allele. By placing adjacent mutational site, difference electrophoretic mobility between loop-hybrids mutated DNA distinguishable native polyacrylamide gel. The also modified in-frame mutations adenocarcinomas. In addition, adapted B-type Raf kinase (BRAF) at V600 Ras-oncogene (NRAS) Q61, commonly found thyroid carcinomas. Our detection system, designated loop-hybrid shift assay sensitive enough comprising 7.5% total DNA. As straightforward technique, may be useful for molecular diagnosis certain types clinical cancers. Other applications are discussed.