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

作者：Zhu, De Bin; Xing, Da^*; Tang, Ya Bing

来源：Chinese Chemical Letters, 2007, 18(7): 869-871.

DOI：10.1016/j.cclet.2007.05.002

• allele-specific amplification
• electrochemiluminescence
• single nucleotide polymorphism
• K-ras oncogene

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)(3)(2+) (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.

• 出版日期2007-7
• 单位华南师范大学