激光生物学报2007，Vol.16Issue(2)：234-237,4.
高灵敏度电化学发光PCR方法检测基因点突变研究
Study on the Highly Sensitive ECL-PCR Method for Point Mutation Detection
摘要
Abstract
Recently, we have developed an electrochemiluminescence-polymerase chain reaction (ECL-PCR) method for point mutation detection. In this method, the target gene was amplified by a Ru( bpy)2+3 (TBR)-labeled forward and a biotinylated reverse primer, and then followed by digestion with a kind of restriction enzyme, which only cut the wildtype (or mutant) amplicon containing its recognition site. Reaction product was detected by electrochemiluminescence (ECL) assay after adsorption of the resulting DNA duplex to the solid phase. One strand of PCR products carries biotin to be bound on a streptavidin-coated microbead for sample selection. Another strand carries TBR to react with tripropylamine (TPA) and emit light for ECL detection. The method was applied to detect the point mutation in presenilin-1 and H-ras genes. The two genotypes were clearly discriminated. In summary, the ECL-PCR method can be used to detect a point mutation that creates or destroys a restriction site in any gene. It is useful in point mutation detection due to its sensitivity, rapidness, simplicity and safety.关键词
电化学发光PCR/点突变检测/Presenilin-1基因/H-ras癌基因Key words
electrochemiluminescence-polymerase chain reaction/point mutation detection/presenilin-1 gene/H-ras oncogene分类
生物科学引用本文复制引用
朱德斌,邢达..高灵敏度电化学发光PCR方法检测基因点突变研究[J].激光生物学报,2007,16(2):234-237,4.基金项目
This research is supported by the National Natural Science Foundation of China (30600128 （）
60378043 （）
30470494) （）
and The Natural Science Foundation of Guangdong Province (015012 （）
04010394 （）
2004B10401011 ). （）
