Alternaria toxin Alternaria monomethyl ether colloidal gold immunochromatographic test strip
A technology of immunochromatographic test paper and sporol monomethyl ether, which is applied in the fields of food safety, immunocolloidal gold test strips and mycotoxin residue detection, and can solve the problems of time-consuming and laborious, long detection time, expensive equipment and the like.
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Embodiment 1A
[0052] The preparation of embodiment 1AME monoclonal antibody
[0053] 1. AME hapten synthesis
[0054] Dissolve 10mg of AME, 14.5mg of methyl bromobutyrate and 30.2mg of potassium carbonate in 1mL of formamide (DMF) solution, and react with magnetic stirring at 50°C for 2h; Extraction was carried out to obtain a precipitated intermediate product (15 mg); the precipitate was dissolved in a mixture of 0.5 mL methanol and 0.5 mL tetrahydrofuran (THF), then added to an aqueous solution (0.1 mL) containing 1.8 mg lithium hydroxide, and reacted for 5 h at room temperature, The carboxylated AME hapten (40 mg) was obtained. The carboxylation modification process of AME and the NMR and LC-MS characterization spectra are shown in figure 1 , 2 , 3.
[0055] 2. Synthesis of immunogen
[0056] The immunogen of the present invention is obtained by coupling AME hapten with BSA. Specific preparation method: Dissolve 15.8mg of AME hapten in 1.5mL of N,N-dimethylformamide (DMF); add 13.7...
Embodiment 2A
[0071] The preparation of embodiment 2AME colloidal gold immunochromatography test strip
[0072] 2.1 Preparation of colloidal gold
[0073] Take 99mL of ultrapure water and 1mL of 1% trichloroauric acid aqueous solution, mix well, heat to boil with magnetic stirring, add 1.8mL of 1% trisodium citrate solution, and continue to heat to boil. When the color of the solution changes from light yellow to blue-black, and finally to wine red, continue heating and boiling for 10 minutes. When the temperature of the prepared colloidal gold solution drops to room temperature, the prepared colloidal gold solution is constant to 100mL. Finally, the prepared colloidal gold solution was placed in a cleaned glass bottle and stored at 4°C for future use.
[0074] 2.2 Preparation of colloidal gold-AME antibody complex
[0075] Get the colloidal gold solution 2mL that prepared particle size is 18nm, with 0.1M K 2 CO 3 Adjust its pH to 8.0; then add 200 μL of AME monoclonal antibody (obtain...
Embodiment 3A
[0086] Embodiment 3AME colloidal gold immunochromatography test strip detection principle
[0087] 1. Detection method: Take the sample to be tested with a disposable straw, and drop 3-4 drops (about 70 μL) on the sample pad.
[0088] 2. Result judgment:
[0089] Colloidal gold test strip test results judgment see Figure 5 .
[0090] Positive: The T line does not show color, but the C line shows color, indicating that the concentration of AME in the sample is higher than the detection limit, and the test result is positive;
[0091] Negative: Both the T line and the C line are colored, indicating that the concentration of AME in the sample is lower than the detection limit, and the test result is negative;
[0092] Invalid: The C line does not develop color, indicating that the test strip has deteriorated and failed, and the test result is invalid.
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Abstract
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