Method for detecting target numerator based on nucleic acid aptamer and PCR amplification

Abstract

The invention discloses a PCP (aptamer based polymerase chain reaction, A-PCR) based on an aptamer, which relates to a high-sensitivity technology for detecting a micro-target matter. In the technology, the high specificity shown in the integration of a nucleic acid aptamer and a ligand and the high sensibility of the aptamer reacted with a polymerase chain are organically combined; the character of the technology is a method which augments a section of DNA reporter molecule through PCP so as to amplify the combination rate of the aptamer and the ligand, thereby detecting a target ligand. The ligand PCR is one of the most sensitive detecting modes at the moment and can detect a single target molecule in theory. Compared with immuno PCR, the technology has the characteristics of simple operation and process, less affection factor, more reliable result, and the like; the sensibility is improved by 10<2> to 10<8> times than the existing ELISA method; the technology can be broadly applied to the micro detection of protein molecules and small molecules.

Description

technical field [0001] The invention relates to a method for identifying and combining target molecules in a sample with nucleic acid molecules, and using PCR method to amplify to quantify and detect target molecules. Background technique [0002] Nucleic acid aptamer (aptamer) refers to the oligonucleotide fragments that can specifically bind proteins or other small molecular substances screened by in vitro screening technology (Systematic evolution of ligand by exponential enrichment, SELEX), and have a certain effect on the binding ligands. Strict recognition ability and high affinity. Since 1990, Tuerk et al. successfully screened phage T for the first time using SELEX technology. 4 Since the RNA-type aptamer of DNA polymerase, as of June 8, 2004, according to the report of the Aptamer Database website, various laboratories have successfully screened out 2,882 nucleic acid aptamers through this technology, and many aptamer sequences for patent protection Not counted ye...

AI Technical Summary

Problems solved by technology

However, there are many factors affecting immuno-PCR, such as linker molecules, selection of display system, concentration of DNA reporter molecules, number of PCR cycles, etc., which all affect its sensitivity and specificity. At present, the technology is not yet fully mature, which limits its further development. application

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