K-ras point mutation detection in lung cancer: comparison of two approaches to somatic mutation detection using ARMS allele-specific amplification.

摘要: Background: The use of sensitive molecular techniques to detect rare cells in a population is increasing interest the pathologist, but detection limits often are poorly defined any given assay. We combined approaches real-time quantitative PCR with ARMSTM allele-specific amplification novel assay for detecting mutant K- ras sequences clinical samples. Methods: ARMS reactions were used seven commonly occurring mutations oncogene. These produce amino acid changes codon 12 (Gly Ala, Arg, Asp, Cys, Ser, or Val) and 13 Asp). A control reaction was measure total amount amplifiable sequence sample so that ratio wild-type could be measured. Quantitative data confirmed selection samples by an independent cloning sequencing method. analyze 82 lung tumor DNA samples. Results: detected 44% adenocarcinomas, which equivalent frequencies reported literature using ultrasensitive techniques. Forty-six percent squamous carcinomas also positive. 0.04–100%. Conclusions: homogeneous, addition being only step before results generated. nature can potentially define analytical sensitivity necessary specified diagnostic application (or other) point mutation detection.