Reagent for quantitative detection of Beta-receptor stimulant through Europium chelate latex time-resolved immunochromatographic assay
A time-resolved fluorescence and receptor agonist technology, applied in the field of β-receptor agonists and veterinary drug residues, can solve the problems of high sensitivity, expensive, complicated process, etc., to overcome low sensitivity, easy storage and transmission, quantitative detection The effect of the method
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Embodiment 1
[0037] Example 1 Preparation of chromatography detection reagent strip / card (eg clenbuterol hydrochloride)
[0038] 1) Packed with Eu 3+ Chelate fluorescent latex-labeled clenbuterol hydrochloride antibody: take 100ul of 10% 200nm europium latex microspheres with carboxyl groups on the surface (Thermo Fisher Company), add 900ul of 0.1M MES pH6.1 for washing, centrifuge at 13000rpm for 30 minutes, discard clear. Add 1ml 0.1MMES pH6.1 for ultrasonic resuspension, add NHS (N-hydroxysuccinimide, Thermo Fisher) 1.2mg, EDC.HCl (1-ethyl-(3-dimethylaminopropyl) carbonyl Diimine hydrochloride, Thermo Fisher) 2 mg, activated reaction at room temperature for 1 hour, centrifuged at 13000 rpm for 30 minutes, and discarded the supernatant. Ultrasonic resuspend with 1ml 0.1M MES buffer pH6.1, add 0.2mg clenbuterol hydrochloride antibody and mix well, react at room temperature (25-30°C) for 2 hours. The labeled latex was centrifuged at 13,000 rpm for 30 minutes, and the supernatant was dis...
Embodiment 2
[0050] Example 2 sample detection
[0051] A. Standard curve establishment
[0052] Prepare 1.0ml of 0.01M pH7.2 PBS solution of clenbuterol hydrochloride with a concentration of 6.4ng / ml, take 500ul and carry out doubling dilution with PBS to obtain different concentrations of clenbuterol hydrochloride solutions (3.20, 1.60, 0.80, 0.40, 0.20, 0.10, 0.05ng / ml), and PBS was used as blank control. Take 100ul standard product directly into the labeled microporous container, dissolve the labeled fluorescent latex particles attached to the wall of the container, mix well, add dropwise on the sample pad of the test strip or test card, react at room temperature for 5 minutes, and then put the test strip Insert a fluorescence reader to detect the fluorescence values in the T and C areas, and the results are shown in the table below.
[0053]
[0054] B. Data processing
[0055] After obtaining the fluorescence values of the T-line and C-line, calculate the T / C ratio, take th...
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