Method for realizing visual nucleic acid testing by composite magnetic beads

Abstract

The invention discloses a method for realizing visual nucleic acid testing by composite magnetic beads and relates to the technical field of nucleic acid testing. The method comprises steps as follows: proteinase K is added to a to-be-tested sample, a lysate and extracting magnetic beads are added and mixed uniformly for a reaction, then the magnetic beads are adsorbed in a magnetic environment, acleaning solution is added, uniform mixing is carried out, the magnetic beads are re-suspended and adsorbed magnetically, liquid is removed, and after drying is performed, the magnetic beads are re-suspended by an eluent; an isothermal amplification reaction solution and a biotin modified primer are added, isothermal amplification is carried out, and an amplification product is obtained; amplicon-targetedly-enriching magnetic beads modified with avidin and FAM-DNA-NH2-fluorescence probe magnetic beads are added, amplicon magnetic beads are formed after incubation, the magnetic beads are magnetically adsorbed, a supernatant is removed, and mixed magnetic beads are obtained by drying; and a Cas12a+crRNA premixed solution is added, re-suspending is carried out, and after a reaction, the magnetic beads are magnetically adsorbed, and observation is performed under blue light. According to the method, nucleic acid extraction, amplification and detection are completed in a tube, reaction solutions of different conditions are replaced in the processes, free switching of different reaction systems can be realized by extraction samples, primers and probes combined on the magnetic beads, andtherefore, the accurate qualitative detection is completed.

Description

technical field [0001] The invention relates to the technical field of high-strength fiber materials, in particular to a nucleic acid visualization inspection method using composite magnetic beads. Background technique [0002] Nucleic acid detection of biological samples is widely used, including disease diagnosis, scientific research, pathogen diagnosis, species identification, etc. At present, mature technologies mainly rely on nucleic acid extraction and PCR amplification of gene-targeted nucleic acid acquisition, sequencing, electrophoresis, and fluorescence quantitative analysis of nucleic acid It is not difficult for professional and technical personnel to operate and implement such methods in the laboratory. However, for non-professionals or institutions or individuals lacking relevant professional equipment, the qualitative detection of nucleic acids is quite difficult, and the previous technology is difficult to achieve. [0003] In addition to nucleic acid detecti...

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Problems solved by technology

[0002] Nucleic acid detection of biological samples is widely used, including disease diagnosis, scientific research, pathogen diagnosis, species identification, etc. At present, mature technologies mainly rely on nucleic acid extraction and PCR amplification of gene-targeted nucleic acid acquisition, sequencing, electrophoresis, and fluorescence quantitative analysis of nucleic acid This method is not difficult to operate and implement by professional technicians in the laboratory, but for non-professionals or institutions or individuals lacking relevant professional equipment, the qualitative detection of nucleic acid is quite difficult, and the previous technology is difficult to achieve

[0003] In addition to nucleic acid detection, antibody detection technology is also a major means of pathogen detection. At present, there are colloidal gold reagents that can complete antibody detection. The method is simple and does not require equipment. However, this method has obvious disadvantages compared with nucleic acid detection, such as pathogen infection. In the early stage, the characterization of proteins or antibodies is carried out. Due to the low abundance, false negatives are prone to occur, and there are interferences from other antibodies or pathogens. This type of detection is also prone to false negatives.

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