Abstract
An electrochemiluminescence-polymerase chain reaction (ECL-PCR) method for rapid detection of K-ras point mutation was developed. Briefly, the target gene was amplified by a Ru(bpy) 3 2+-labeled forward primer and a biotin-labeled reverse primer, and then followed by digestion with a restriction enzyme, MvaI, which only cut the wild-type amplicon containing its cutting site. Digested product was detected by the ECL assay after adsorption of the resulting DNA duplex to the solid phase. Thirteen pancreatic cancer samples were analyzed by the ECL-PCR assay. The positive rate of K-ras point mutation was 92.3%. The ECL-PCR method is useful in point mutation detection owing to its sensitivity, rapidness, simplicity, and safety. It can be used to detect a point mutation that creates or destroys a restriction site in any gene.
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