Carbofuran assay determination kit and application thereof
A kit, the technology of Carboweil, is applied in the field of food safety detection immunoassay to achieve the effects of improved sensitivity, easy operation and production, and good reproducibility
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Embodiment 1
[0047] Preparation method of carbofuran analysis and determination kit
[0048] S11, Preparation of carbofuran enzyme-labeled colloidal gold composite probe
[0049] Gold Nanoparticle Synthesis Method
[0050] Add 98mL of deionized water to the siliconized beaker respectively, and then add 2mL of 50mM chloroauric acid stock solution respectively, so that the concentration of chloroauric acid is 1mM, stir on a magnetic stirrer at 1000rpm / min, heat at 250°C, continue to boil after the liquid boils Boil for 2 minutes; absorb 10mL of 38.8mM trisodium citrate, quickly add it to the beaker at one time, keep the stirring speed and heating temperature constant, and continue to boil for 6 minutes. It can be found that the color of the solution gradually changes from colorless to purple, and then into wine red. According to the different reducing doses of trisodium citrate added, it finally turns into different colors. After the liquid is cooled, deionized water is used to restore the ...
Embodiment 2
[0074] Kit use method of the present invention
[0075] The specific operation of the Carbofuran enzyme-labeled colloidal gold composite probe amplification immunoassay kit prepared in the above implementation case 1 is as follows:
[0076] S21. Take out the kit from the refrigerator at 4°C, and equilibrate at room temperature for 10 minutes;
[0077] S22. Carbofuran standard substances (10.0ng L -1 -40 μg L -1 ) and 20 μL of each sample obtained after pretreatment, set up a repetition, then add 20 μL of magnetic nanoprobes to each well, fully oscillate and mix with a micro shaker, and then place the temperature at 37 ° C and a relative humidity of 70% constant temperature Incubate for 1 h in a constant humidity incubator;
[0078] S23, washing with PBST solution three times, dehybridization to obtain a competition complex;
[0079] S24. Add 50 μL of the reaction substrate TMB to the competition reaction complex, react at 37°C for 15 minutes, then add 50 μL of stop solution ...
experiment example 1
[0082] The methodological examination result of kit of the present invention
[0083] The kit prepared in Example 1 was identified according to the common verification procedures in this field, and the results are shown in Table 1.
[0084] The methodological testing result of test kit of the present invention of table 1
[0085]
[0086] The above results show that the accuracy, specificity, precision, sensitivity and stability of the "Carbofuran Colloidal Gold Composite Probe Combined with HRP Immunoassay Assay Kit" are fully qualified.
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