Citation: De Bin Zhu, Da Xing, Ya Bing Tang. Allele-specific amplification and electrochemiluminescence method for single nucleotide polymorphism analysis[J]. Chinese Chemical Letters, ;2007, 18(7): 869-871. doi: 10.1016/j.cclet.2007.05.002 shu

Allele-specific amplification and electrochemiluminescence method for single nucleotide polymorphism analysis

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, South China Normal University, Guangzhou 510635, China

Received Date: 27 December 2006

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 30600128 30670507 30470494) and the Natural Science Foundation of Guangdong Province (No. 015012).

A new approach combined the specificity of allele-specific amplification (ASA) with the sensitivity of electrochemiluminescence (ECL) assay for single nucleotide polymorphism (SNP) analysis was proposed. Briefly, target gene was amplified by a biotin-labeled allele-specific forward primer and a Ru(bpy)₃²⁺ (TBR)-labeled universal reverse primer. Then, the amplicon was captured onto streptavidin-coated paramagnetic beads through biotin label, and detected by measuring the ECL signal of TBR label. Different genotypes were distinguished according to the ECL values of the amplicons by different genotypic primers. K-ras oncogene was used as a target to validate the feasibility of the method. The experiment results show that the different genotypes can be clearly distinguished by ASA-ECL assay. The method is useful in SNP analysis due to its sensitivity, safety, and simplicity.
- Allele-specific amplification,
- Electrochemiluminescence,
- Single nucleotide polymorphism,
- K-ras oncogene
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