Abstract

A highly sensitive electrochemiluminescence-polymerase chain reaction (ECL-PCR) method for K- ras point mutation detection is developed. Briefly, K- ras oncogene was amplified by a Ru(bpy) 3 2+ (TBR)-labeled forward and a biotin-labeled reverse primer, and followed by digestion with MvaI restriction enzyme, which only cut the wild-type amplicon containing its cutting site. The digested product was then adsorbed to the streptavidin-coated microbead through the biotin label and detected by ECL assay. The experiment results showed that the different genotypes can be clearly discriminated by ECL-PCR method. It is useful in point mutation detection, due to its sensitivity, safety, and simplicity.

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