Rolling circle amplification-based colorimetric assay method for target nucleic acids or proteins

Abstract

The invention relates to a rolling circle amplification-based colorimetric assay method for target nucleic acids or proteins. The method mainly utilizes the hybridization between nucleic acids and antigen-antibody binding to indirectly fix the target nucleic acids or proteins to be assayed onto magnetic beads, rolling circles are added, a rolling circle primer on hybrid is utilized to carry out constant-temperature rolling circle amplification, and finally, the chromogenic reaction of nanogold is utilized to assay constant-temperature amplified macromolecular product in order to achieve the purpose of relatively assaying the target nucleic acids and proteins. By integrating rolling circle amplification and the chromogenic reaction of nanogold, the method not only amplifies a large quantity of micro-sample signals, but also realizes the rapid, visual assay of results, avoids the usage of a PCR (Polymerase Chain Reaction) instrument and other expensive instruments, and decreases the requirement on the experimental hardware condition. The method is expected to be applied in bedside rapid diagnosis and the field rapid assay of microorganisms in food safety and the environment.

Description

technical field [0001] The invention relates to a colorimetric detection method for target nucleic acid or protein based on rolling circle amplification, which belongs to the detection field of biological macromolecules. Background technique [0002] Rapid, specific, and high-throughput biomolecular detection methods are of great significance to the diagnosis of pathogenic microorganisms and genetic diseases in the fields of medical care and biosafety. The highly sensitive detection of low-concentration biomolecules directly from samples has broad and potential applications in a large number of fields such as laboratory medicine, pathology, epidemiology, environmental and food testing. At present, the most widely used amplification method is mainly polymerase chain reaction (PCR). PCR technology can realize the amplification of 10 DNA molecules, and has high sensitivity and specificity. However, its main disadvantage is that it needs to use complex The equipment and the rea...

AI Technical Summary

Problems solved by technology

At present, some detections based on rolling circle amplification are relatively complicated and require expensive instruments, such as chip detection (whole genome rolling circle Amplified oligonucleotides, patent number: 200480027696.1) and Raman spectroscopy detection (Biomolecule analysis by rolling circle amplification and SEBS detection, patent number: 200480001922.9), obviously these methods are not suitable for rapid detection on the spot

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