Multi-target simultaneous detection method based on latex microsphere ultraviolet absorption spectrum peak area integration

A technology of latex microspheres and area integration, which is applied in the field of biochemical analysis, can solve problems such as failure to meet amplification requirements, inactivation of biorecognition molecules, and weakening signal gaps, etc., to avoid baseline background interference, high accuracy, and small range Effect

Pending Publication Date: 2022-04-22
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a major problem in introducing click reaction as an effective signal amplification system is the appropriate dosage. Too little dosage will not meet the amplification requirements, and too much dosage will lead to the inactivation of biorecognition molecules, which will easily weaken the signal gap and reduce sensitivity.

Method used

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  • Multi-target simultaneous detection method based on latex microsphere ultraviolet absorption spectrum peak area integration
  • Multi-target simultaneous detection method based on latex microsphere ultraviolet absorption spectrum peak area integration
  • Multi-target simultaneous detection method based on latex microsphere ultraviolet absorption spectrum peak area integration

Examples

Experimental program
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Effect test

Embodiment 1

[0083] Multiple Quantitative Detection of Example 1 Deoxynivalenol (DON), Aflatoxin (AFB) and Ochratoxin (OTA)

[0084] Simultaneous detection of vomitoxin, aflatoxin and ochratoxin by competitive immunoassay, the schematic diagram is as follows Figure 9 As shown, magnetic particles with a particle size of 1000nm were used as magnetic separation carriers, and antibodies corresponding to vomitoxin, aflatoxin, and ochratoxin were coupled to the surface; 99nm, 500nm, and 1050nm PS microspheres were used as signal probes, respectively. The complete antigen coupled with three targets, when the target exists, the complete antigen will compete with the target for the antibody on the surface of the carrier to form nano-magnetic particles-antibody-target, nano-magnetic particles-antibody-complete antigen-PS microparticles Sphere complexes and unreacted signaling probe solutions. Therefore, the content of the target directly affects the number of signal probe-carrier complexes formed....

Embodiment 2

[0096] Example 2 Multivariate signal amplification method for deoxynivalenol (DON), aflatoxin (AFB) and ochratoxin (OTA)

[0097] Simultaneously improve the sensitivity of analyzing vomitoxin, aflatoxin and ochratoxin, the schematic diagram is as follows Figure 11 As shown, magnetic particles with a particle size of 1000nm are used as magnetic separation carriers, and antibodies corresponding to vomitoxin, aflatoxin, and ochratoxin are coupled on the surface; ;Using 99nm, 500nm, 1050nm PS microspheres as signal probes, respectively coupled with click reagent II corresponding to click reagent I, when the target exists, the complete antigen will compete with the target for the antibody on the surface of the carrier to form nano Magnetic particle-antibody-target object, magnetic nanoparticle-antibody-complete antigen-click reagent I-click reagent II-PS microsphere complex and signal probe solution not involved in the reaction. Therefore, the content of the target directly affec...

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Abstract

The invention discloses a latex microsphere ultraviolet absorption spectrum peak area integral-based multi-target simultaneous detection method, which is characterized in that nano magnetic particles are used as magnetic separation carriers, and biological recognition molecules A corresponding to different targets are coupled on the surfaces of the nano magnetic particles to serve as immune magnetic particles; latex microspheres are used as signal probes, and corresponding biological recognition molecules B are coupled to the surfaces of the latex microspheres; the immune magnetic particles, the signal probe and a to-be-detected target object are subjected to a sufficient immune reaction, and a mixed solution is obtained; carrying out magnetic separation, carrying out full-wavelength scanning on the signal probe solution which does not participate in the reaction, and selecting a corresponding wavelength range to carry out peak area integration. According to the method, the sensitivity and the accuracy are remarkably improved, and accurate analysis and quantification of each signal probe can be realized only by carrying out peak area integration on the ultraviolet absorption spectrum in a specific range. For substances with higher analysis sensitivity requirements, various click reactions are adopted to further amplify optical signals, and the dosage of a click reagent is optimized, so that the purpose of simultaneously amplifying various target object signals is achieved.

Description

technical field [0001] The invention belongs to the field of biochemical analysis, in particular to a multi-target simultaneous detection method based on latex microsphere ultraviolet absorption spectrum peak area integration. Background technique [0002] Optical biosensing technology is a microanalysis technology based on the generation of various optical signals for detection purposes. It has been widely used because of its simple operation, high sensitivity and low cost. At present, the widely used biosensing methods mainly include enzyme-linked immunosorbent assay, immunoturbidimetric assay, optical biosensing method based on nano-gold and magnetic relaxation sensing method based on nano-magnetic particles. [0003] ELISA colorimetric method has high specificity and is the most widely used, but it takes a long time to detect and has low sensitivity; compared with ELISA, immunoturbidimetric method is simple to operate and low in cost, but consumes a lot of antibodies and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/33
CPCG01N33/54313G01N33/54386G01N21/33
Inventor 陈翊平周翠云潘世兴鲁鹏王治潘
Owner HUAZHONG AGRI UNIV
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