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Colloidal gold quenching quantum dot immunochromatographic test strip for detecting zearalenone and preparation method of test strip

A technology of zearalenone and immunochromatographic test paper, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of complicated sample processing steps, unsuitable for popularization and use, and the need for derivation, etc., and achieves simple operation and detection Fast time, high specific effect

Inactive Publication Date: 2017-10-10
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of this method is that it requires expensive instruments and professional personnel to operate, the sample processing steps are cumbersome, derivation is required, and the recovery rate is low, so it is not suitable for widespread promotion and use.

Method used

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  • Colloidal gold quenching quantum dot immunochromatographic test strip for detecting zearalenone and preparation method of test strip
  • Colloidal gold quenching quantum dot immunochromatographic test strip for detecting zearalenone and preparation method of test strip
  • Colloidal gold quenching quantum dot immunochromatographic test strip for detecting zearalenone and preparation method of test strip

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Experimental program
Comparison scheme
Effect test

Embodiment l

[0051] Embodiment 1 (preparation embodiment)

[0052] (1) Purification of Zearalenone Ascites

[0053] The zearalenone ascites used in this experiment was self-made by the laboratory, and the antibody in the ascites was purified by the octanoic acid-ammonium sulfate method. The specific operation steps are as follows:

[0054] 1) Take 1 mL of ascitic fluid, add 2 mL of acetate buffer solution (0.06mol / L, pH 4.0), mix well and adjust the pH to 4.8 with 1 mol / L NaOH solution.

[0055] 2) Slowly add 33 μL of n-octanoic acid while stirring, and place it at 4° C. for 2 hours.

[0056] 3) Centrifuge at 15,000 rpm for 30 min at 4°C, discard the precipitate, and collect the supernatant.

[0057] 4) Add PBS solution (0.1mol / L PBS, pH 7.4) about 1 / 10 of the supernatant volume to the supernatant, and adjust the pH to 7.4 with 1mol / L NaOH solution.

[0058] 5) Slowly add ammonium sulfate to the solution in an ice bath state, adding 0.277g per milliliter of the solution, and stir for 30...

Embodiment 2

[0066] Embodiment 2 (preparation embodiment)

[0067] Assembly and preparation method of quantum dot fluorescence quenching immunochromatographic test strip

[0068] 1. Assembly of test strips:

[0069] The test strip of the present invention is composed as follows: sample pad 1, nitrocellulose membrane 2, water-absorbing pad 3 and PVC plate 6, on the PVC plate 6, sample pad 1, nitrocellulose membrane 2, water-absorbing pad are adhered in sequence 3. The nitrocellulose membrane 2 is coated with a detection line 4 composed of a zearalenone antigen and a quality control line 5 composed of a goat anti-rabbit secondary antibody, respectively.

[0070] 2. Preparation of quantum dot-chicken ovalbumin conjugate:

[0071] Quantum dots-chicken ovalbumin (QDs-OVA) conjugates were prepared by activated ester method.

[0072] (1) Coupling reaction. Take 50 μL quantum dots in a centrifuge tube, add 0.6 mg chicken ovalbumin and 23 μL EDC solution (10 mg / ml), mix well, and supplement the...

Embodiment 3

[0085] Embodiment 3 (application embodiment)

[0086] 1. How to use quantum dot-labeled immunochromatographic test strips

[0087] Pretreatment of Grain Samples

[0088] Put 5g of grain samples into a 50mL centrifuge tube after grinding, add 5mL of 70% methanol solution, mix at high speed for 5min, supplement the volume with phosphate buffer (pH 7.5) to 30mL, and centrifuge at 5000g for 5min at 4°C. Aspirate the supernatant and use it directly for test strip detection.

[0089] 2. Detection steps

[0090] Use a pipette gun to draw 200 μL of the sample extract to be tested into the safety tube, add 8 μL of AuNPs-ZEN-Ab, mix well, add dropwise to the sample hole of the test strip, and observe the result after 15 minutes.

[0091] 3. Result judgment

[0092] Such as figure 2 As shown, if the fluorescence of the detection line of the test strip of the sample to be tested disappears, it is judged as a negative sample, that is, it does not contain zearalenone; If the color of...

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Abstract

The invention provides a colloidal gold quenching quantum dot immunochromatography test strip for detecting zearalenone, comprising a sample pad 1, a nitrocellulose membrane 2, a water-absorbing pad 3 and a PVC backboard, characterized in that, A sample pad 1, a nitrocellulose membrane 2, and a water-absorbing pad 3 are adhered to the PVC backboard in sequence; the nitrocellulose membrane 3 is coated with zearalenone antigen and quantum dot-chicken ovalbumin respectively. The detection line 4 formed by the conjugate and the quality control line 5 formed by the quantum dot-chicken ovalbumin conjugate. The invention also discloses a preparation method of the test strip. The invention has the following prominent advantages: 1. High specificity and good sensitivity; 2. Low detection cost. 3. Easy to operate.

Description

technical field [0001] The invention relates to a method for rapidly detecting zearalenone in grain food, in particular to a colloidal gold quenching quantum dot immunochromatographic test strip for detecting zearalenone and a preparation method thereof. Background technique [0002] Zearalenone is a phenolic mycotoxin mainly produced by Fusarium moniliforme, Fusarium tritinarum, and Gibberella zearaenone. Zearalenone has the effect of estrogen, mainly acting on the reproductive system, affecting the estrogen level of poultry and livestock, causing diseases of the reproductive system of livestock, mainly manifested as genital swelling [6], hyperemia, stillbirth and abortion, mental depression, Loss of appetite, abdominal pain, diarrhea and other problems, but also have a certain toxic effect on the nervous system, heart, kidneys, liver and lungs. At present, there is no specific drug treatment for animal zearalenone poisoning, and the direct cause of animal zearalenone pois...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/558
CPCG01N33/54313G01N33/558
Inventor 生威张燕王俊平王硕刘冰李诗洁
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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