Method for rapidly and sensitively detecting MicroRNA based on water-soluble fluorescent polymer

Abstract

The invention discloses a method for rapidly and sensitively detecting MicroRNA based on a water-soluble fluorescent polymer. The method comprises the steps that probe SH-DNA is combined with gold magnetic particles to obtain mercapto-labeled gold magnetic particles, and acryloyl fluorescein, probe Acrydite-DNA and acrylamide are copolymerized to obtain the water-soluble fluorescent polymer; MicroRNA is hybridized with DNA and SH-DNA of the water-soluble fluorescent polymer at the same time; the fluorescence intensity of supernatant is measured after magnetic separation, de-hybridization and magnetic separation again, wherein the higher the fluorescence intensity is, the greater the concentration of MicroRNA is. The method adopts the water-soluble fluorescent polymer to realize the multi-stage amplification of detection signals, and the surfaces of the gold magnetic particles are modified with SH-DNA sequences, thus the separation of hybrid products is realized and the detection operation is simplified. The method does not need to amplify the target microRNA, and is low in cost, low in professional requirement, simple to operate, fast in analysis speed, high in selectivity and low in detection limit.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for rapidly and sensitively detecting MicroRNA based on a water-soluble fluorescent polymer. Background technique [0002] MicroRNA (MicroRNA) is a non-coding small molecule RNA with a base number of 19-23, which widely exists in animals, plants and viruses. MicroRNA is closely related to various physiological processes of organisms, including growth and development, immune Defense, hematopoietic processes, organ formation, cell proliferation and apoptosis, fat metabolism, tumor development, etc. Understanding the regulation of microRNAs and the relationship between microRNA expression and human disease has aroused great interest among researchers. Calin et al. first discovered that the expression of MicroRNA is closely related to the occurrence of human tumors, and has found that the expression of MicroRNA changes in many types of tumors, and some MicroRNA can ...

AI Technical Summary

Problems solved by technology

Among them, the two technologies of RT-PCR and LCR are variable temperature reactions, which need to rely on a sophisticated thermal cycler. If the temperature control is not appropriate during the heating and cooling process, the amplification will fail.

Moreover, these techniques all require the participation of enzymes, that is, an amplification technique requires an enzyme or even a combination of multiple enzymes to catalyze, otherwise the amplification reaction cannot occur, and these amplification techniques also require complex primer and probe sequences. Sophisticated design and high professional requirements for experimental operations

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