Turn-on test strip for detecting T-2 toxin and preparation method thereof

A technology of T-2 and test strips, applied in the direction of biological testing, measuring devices, material inspection products, etc., to achieve the effects of large Stokes shift, improved sensitivity, and high photobleaching resistance

Pending Publication Date: 2021-11-02
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, using colloidal carbon as a fluorescence quencher and time-resolved fluorescent microspheres as a fluorescence donor material, a test strip with high sensitivity, low cost and fast detection speed is prepared, which has potential application prospects. to report

Method used

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  • Turn-on test strip for detecting T-2 toxin and preparation method thereof
  • Turn-on test strip for detecting T-2 toxin and preparation method thereof
  • Turn-on test strip for detecting T-2 toxin and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0029] The preparation of test strip described in embodiment 1:

[0030] (1) Wash time-resolved fluorescent microspheres: take 0.1mL time-resolved fluorescent microspheres (1%), dilute with 0.9mL 50mmoL / L pH 6.0 MES buffer, mix well, centrifuge at 8500 rpm for 15min, discard the supernatant, and press Wash again with the same steps. Finally, it was resuspended in 1.0 mL of 50 mmoL / L pH 6.0 MES buffer.

[0031] (2) Activation of time-resolved fluorescent microspheres: 5.0 μL of 20 mg / mL EDC and 20.0 μL of 20 mg / mL NHS were sequentially added to the solution obtained in (1), and incubated at room temperature for 30 min. Centrifuge the activated time-resolved fluorescent microspheres at 8500 rpm for 15 min, discard the supernatant, and resuspend the pellet in 1.0 mL of 50 mmoL / L pH 6.0 MES buffer.

[0032] (3) Preparation of time-resolved fluorescent microsphere-labeled BSA bovine serum albumin complex: add 20 mg / mL 200 μL of BSA bovine serum albumin to 0.1 mL of activated 10 m...

Embodiment 2

[0036] The establishment of embodiment 2 detection method

[0037] 1. The detection method and principle of the turn-on test strip used to detect T-2 toxin

[0038] The present invention provides turn-on type test strips to quantitatively detect the content of T-2 toxin in the buffer by using the competition method:

[0039] Add 5 μL of the quencher colloidal carbon-labeled antibody complex to the microwell plate, and then add 200 μL of the buffer to be tested. The T-2 toxin in the buffer is first combined with the T-2 toxin in the quencher colloidal carbon-labeled antibody complex The monoclonal antibody binding of the test strip is placed in the microwell plate, and the sample pad is continuously chromatographed. The complex moves slowly on the NC membrane by means of chromatography, and does not combine with the fluorescent donor in the detection line area. , the fluorescence in the detection line area is not quenched, the colloidal carbon-labeled antibody-coupled T-2 toxi...

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Abstract

The invention discloses a turn-on type test strip for detecting T-2 toxin and a preparation method of the turn-on type test strip, and belongs to the technical field of rapid food detection. The test strip consists of a bottom plate, a sample pad, an absorption pad and a nitrocellulose membrane (NC membrane). The method comprises the following steps: time-resolved fluorescent microspheres is selected as a fluorescent material, the time-resolved fluorescent microspheres is coupled with BSA (Bovine Serum Albumin), and a prepared compound is fixed on a test strip to serve as a fluorescent donor; a colloidal carbon labeled antibody compound is used as a quencher, a mixture of a time-resolved fluorescent microsphere labeled BSA bovine serum albumin compound and a T-2 antigen is sprayed on an NC membrane to serve as a detection line, a goat anti-mouse IgG antibody is sprayed to serve as a quality control line, and the turn-on type lateral flow chromatography detection test strip is assembled. The test strip can be used for on-site detection of T-2 toxin, increases the detection sensitivity and the detection speed, is suitable for large-scale application and batch production, and has good application and development prospects.

Description

technical field [0001] The invention relates to a turn-on test strip, in particular to a turn-on test strip for detecting T-2 toxin and a preparation method thereof, belonging to the technical field of rapid food detection. Background technique [0002] T-2 toxin is a sesquiterpene compound, which is widely distributed in nature and is mainly produced by Fusarium trieinetum and Fusarium poae. According to statistics from the Food and Agriculture Organization of the United Nations, it pollutes about 25% of agricultural products in the world every year. One of the mycotoxins. T-2 toxin is one of trichothecene toxins and the most toxic toxin of trichothecenes. It can not only cause acute poisoning such as nausea, dizziness, vomiting, abdominal distention and pain, chills and diarrhea, but also inhibit The synthesis of DNA, RNA and protein has strong genotoxicity, immunotoxicity and blood toxicity. The nutritional and economic direct losses of agricultural products caused by t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/543G01N33/533G01N33/53
CPCG01N33/577G01N33/558G01N33/54313G01N33/533G01N33/5308
Inventor 张西亚丁明月党梦毛烨炫黄现青谢新华宋莲军王田林张彤彤李子哲王尤一
Owner HENAN AGRICULTURAL UNIVERSITY
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