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Sensitivity detection method for fumonisin B1

A technology for sensitive detection of fumonisins, which can be used in measuring devices, biological tests, and material inspection products. Enzyme-catalyzed steps, wide range of affinity changes, and simple operation

Active Publication Date: 2017-05-10
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical defect of prior art, provides a kind of anti-fumonisin B 1 Sensitive detection method to address fumonisin B in the prior art 1 Technical issues with low sensitivity of the detection method
[0006] Another technical problem to be solved by the present invention is that for fumonisin B in the prior art 1 The direct competition ELISA method has low detection sensitivity due to the poor luminescent performance of the chromogenic substrate
[0007] Another technical problem to be solved by the present invention is that in the prior art, for fumonisin B 1 The direct competition ELISA method has low detection sensitivity due to the high affinity between the competing antigen and the antibody

Method used

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  • Sensitivity detection method for fumonisin B1
  • Sensitivity detection method for fumonisin B1
  • Sensitivity detection method for fumonisin B1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1. Carboxyl-modified quantum dot fluorescent microspheres

[0057] 10mg of quantum dots modified with oleic acid groups (the excitation wavelength of the quantum dots is 450nm and the emission wavelength is 580nm) was dissolved in 0.5mL of chloroform, and then 15mg of methyl methacrylate and 10mg of 1-maleic anhydride were added After half an hour, the mixture was re-dissolved in 2.5mL of sodium sulfonate aqueous solution with a final concentration of about 3.3mg / mL. The mixture was mixed under ultrasonic conditions, and after mixing, chloroform was removed by rotary evaporation. The water-soluble carboxyl-modified quantum dot fluorescent microspheres are obtained after polar organic solvents, and the quantum dot fluorescent microspheres are separated by centrifugation, and the separated quantum dot fluorescent microspheres are washed three times with ultrapure water. The washed quantum dot fluorescent microspheres were redissolved in ultrapure water and stored at 4°C. ...

Embodiment 2

[0088] fumonisin B 1 A sensitive detection method comprising the following steps:

[0089] 1) Coating antibody on the microtiter plate;

[0090] 2) Prepare fluorescent microspheres labeled with carboxyl-modified quantum dots;

[0091] 3) Combine the product obtained in step 2) with fumonisin B 1 Coupling, that is, to obtain the competing antigen;

[0092] 4) Add the solution to be tested and the competing antigen to the antibody-coated microtiter plate in step 1), react with antigen-antibody binding, and then detect the fluorescence intensity of the microtiter plate.

[0093] On the basis of the above technical solutions, the following conditions are met:

[0094]Step 2) The fluorescent microspheres labeled with carboxyl-modified quantum dots were prepared by the following method: using chloroform as a solvent, preparing oleic acid group-modified quantum dots at a concentration of 15 mg / mL, PMMA at a concentration of 25 mg / mL, The mixed solution with PMAO concentration of...

Embodiment 3

[0103] fumonisin B 1 A sensitive detection method comprising the following steps:

[0104] 1) Coating antibody on the microtiter plate;

[0105] 2) Prepare fluorescent microspheres labeled with carboxyl-modified quantum dots;

[0106] 3) Combine the product obtained in step 2) with fumonisin B 1 Coupling, that is, to obtain the competing antigen;

[0107] 4) Add the solution to be tested and the competing antigen to the antibody-coated microtiter plate in step 1), react with antigen-antibody binding, and then detect the fluorescence intensity of the microtiter plate.

[0108] On the basis of the above technical solutions, the following conditions are met:

[0109] Step 2) The fluorescent microspheres labeled with carboxy-modified quantum dots were prepared by the following method: using chloroform as a solvent, preparing oleic acid group-modified quantum dots at a concentration of 25 mg / mL, PMMA at a concentration of 35 mg / mL, The mixed solution with PMAO concentration of...

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Abstract

The invention provides a sensitivity detection method for fumonisin B1. The method includes: utilizing fluorescent microspheres embedded with quantum dots to replace conventional enzyme carriers to be coupled with fumonisin B1; using a coupling product as a competition antigen to execute direct competition ELISA (enzyme-linked immune sorbent assay). In a technical path, proper quantum dots are selected according to luminescent characteristics, a quantum dot embedding method based on fluorescent microsphere technology is further designed, and on this basis, quantum dot fluorescent microspheres are subjected to BSA embedding and then coupled with fumonisin B1 to obtain the competition antigen with better performance. In the technical scheme, a lot of quantum dots are embedded in the fluorescent microspheres through a high polymer carrier, so that the fluorescent microspheres have higher illumination intensity, and detection sensitivity can be improved effectively; the fluorescent microspheres have large grain size, so that over-high affinity between the competition antigen and an embedding antibody can be lowered to a certain degree, and detection sensitivity is improved.

Description

technical field [0001] The present invention relates to the technical field of antigen detection, and further relates to the antigen detection technology based on fluorescence immunology analysis, in particular to a method against fumonisin B 1 sensitive detection method. Background technique [0002] Fumonisins are a group of mycotoxins mainly produced by Fusarium moniliforme under certain temperature and humidity conditions. An aliphatic chain composed of two carbon atoms and a hydrophilic side chain connected by two ester bonds. Fumonisins exist in the form of FA 1 、FA 2 , FB 1 , FB 2 , FB 3 , FB 4 、FC 1 、FC 2 、FC 3 、FC 4 , FP 1 Fumonisin B 1 (FB 1 ) are the most common and most toxic. Fumonisins can cause leukomalacia in horses, pulmonary edema and pleural effusion in pigs, atherosclerotic changes in primates, and liver cancer in rats. In humans, fumonisins mainly appear in some areas with high corn consumption. Epidemiological studies in some areas have ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/543G01N33/58
CPCG01N33/533G01N33/54313G01N33/588G01N2333/37
Inventor 熊勇华周耀峰黄小林李响敏江湖赖卫华
Owner NANCHANG UNIV
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