High-flux ultra-sensitive double-label time-resolved immunochromatographic test strip and application thereof
A time-resolved fluorescence, test strip technology, applied in analytical materials, biological material analysis, measurement devices, etc., can solve the problem that the sensitivity is difficult to meet people's requirements, and achieve the effects of rapid detection, high sensitivity and good stability
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Embodiment 1
[0075] Example 1: Preparation of aflatoxin B1 and zearalenone double-labeled time-resolved immunochromatographic test strips
[0076] 1. Preparation of time-resolved fluorescent microsphere-conjugated antibody probes
[0077] 1. Time-resolved fluorescent microspheres conjugated to aflatoxin B1 monoclonal antibody
[0078] (1) Add 5 μL of time-resolved fluorescent microsphere solution to 500 μL of 0.2 mol / L borate borax buffer (pH 8.0), vortex and mix well, add 1 μg EDC and 1 μg NHS respectively, and activate on a shaker in the dark 15min.
[0079] (2) After completing step (1), add 8 μg of aflatoxin B1 monoclonal antibody (purchased from Beijing Weideweikang Biotechnology Co., Ltd., catalog number AFB-12A8), and react at room temperature for 2 hours or overnight at 4°C.
[0080] (3) After completing step (2), add 50 μL of 10% BSA solution containing 1% Tween 20, and block at room temperature on a shaker for 2 hours to fully block redundant activation sites on the surface of ...
Embodiment 2
[0100] Example 2: Qualitative and quantitative detection of aflatoxin B1 and zearalenone
[0101] 1. Qualitative detection method of aflatoxin B1 and zearalenone (line elimination method)
[0102] 1. Take 1 g of sample and add 4 mL of 70% (volume fraction) methanol solution. After shaking for 5 minutes, centrifuge at 8000 rpm for 10 minutes. Collect the supernatant and dilute it 10 times with sample diluent to obtain the processed sample. Take 120 μL of the treated sample, add 1 μL of the time-resolved fluorescent microsphere-coupled aflatoxin B1 monoclonal antibody solution prepared in Example 1, and 1 μL of the time-resolved fluorescent microsphere-coupled zearalen prepared in Example 1 The ketone monoclonal antibody solution and 1 μL of the time-resolved fluorescent microsphere-coupled chicken IgY antibody solution prepared in Example 1 were mixed evenly to obtain a mixed solution.
[0103] 2. After completing step 1, incubate the mixed solution for 5 minutes, then add the...
Embodiment 3
[0116] Example 3: Application of double-labeled time-resolved immunochromatographic method
[0117] 1. Preparation of samples to be tested
[0118] Coix seed, Chinese yam, and Campanulaceae that were identified by high-performance liquid chromatography-secondary mass spectrometry (UPLC-MS / MS) as samples that do not contain aflatoxin B1 and zearalenone were processed as follows: the samples were Grind to obtain a ground sample; take 1 g of the ground sample and add 4 mL of 70% (volume fraction) methanol solution, shake for 5 minutes, then centrifuge at 8000 rpm for 10 minutes, collect the supernatant and dilute it 10 times with the sample diluent to obtain the processed sample .
[0119] Add different concentrations of aflatoxin B1 and zearalenone to the treated samples to obtain test sample solutions containing different concentrations of aflatoxin B1 and zearalenone. Taking coix seed as an example, when the addition amount of AFB1 in the sample solution to be tested is 1 μg...
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