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Time-resolved fluorescence immunoassay kit for detecting cyproheptadine

A time-resolved fluorescence and cyproheptadine technology, applied in biological testing, fluorescence/phosphorescence, analytical materials, etc., can solve the problems of cumbersome operation steps, cumbersome operation, low false positive rate, etc. specific effect

Inactive Publication Date: 2020-07-03
ZHENJIANG YITE BIOTECH DEV CO LTD
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  • Application Information

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography (HPLC) has the characteristics of high detection accuracy and low false positive rate; the main determination of high-performance liquid chromatography (HPLC) is that the instrument is expensive, the operation is cumbersome, time-consuming, and the detection cost is high
Gas chromatography-mass spectrometry (GC-MS) method has high sensitivity and low false positive rate; however, the sample of this method needs to be derivatized, which will cause deviations in the experimental results
Liquid chromatography-mass spectrometry (LC-MS) can detect urine, blood, hair and other samples, but similar to the above method, the experimental operation steps are cumbersome and the detection cost is high
There are no patents and literature reports on the time-based fluorescent immunoassay for the detection of cyproheptadine

Method used

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Embodiment

[0023] 1. Preparation of immunogen and coating agent

[0024] Synthesis of the immunogen (cyproheptadine-BSA) of the present invention: Accurately weigh 324mg of cyproheptadine and dissolve it in 2mL N,N-dimethylformamide, add γ-aminobutyric acid solution dropwise under stirring, and stir for 3 Hours, adjust the pH of the reaction solution to about 10. The precipitate was removed by centrifugation. Add the above reaction dropwise to the BSA solution (320mg BSA dissolved in 5mL physiological saline), then add 23mg of N-hydroxysuccinimide (NHS), 45.4mg of N,N-dicyclohexylcarbodiimide (DCC), React overnight at 4°C, remove the precipitate by centrifugation, take the supernatant and dialyze it with phosphate buffered solution (PBS) for 3 days, change the dialysate every 6 hours, freeze-dry the obtained product, and store it at -20°C for later use;

[0025] Synthesis of the coating agent (cyproheptadine-OVA): In the above reaction, after replacing BSA with OVA, the reaction conjug...

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Abstract

The invention discloses a time-resolved fluorescence immunoassay kit for detecting cyproheptadine. The time-resolved fluoroimmunoassay detection kit for detecting cyproheptadine is composed of a porous coating plate, a buffer solution, a cyproheptadine standard substance, a cyproheptadine antibody freeze-dried product, a europium-labeled goat anti-mouse antibody, a washing solution and an enhancement solution. A detection method of the time fluorescence immunoassay kit for detecting cyproheptadine comprises the following steps: (1) preparing an immunogen; (2) preparing a coated antigen; (3) preparing a monoclonal antibody; and (4) pretreating and detecting a sample. The kit and the method are short in detection time, high in average recovery rate, simple in sample pretreatment, wide in application and low in detection cost, can be operated and detected on site, and have the advantages of high detection specificity, small intra-batch and inter-batch difference, high sensitivity, simplicity and quickness in operation, particular suitability for detection of large-batch samples and the like.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a time-resolved fluorescent immunoassay kit for detecting cyproheptadine. Background technique [0002] Cyproheptadine (CYP) is a human drug, which is clinically used to treat diseases such as dermatitis caused by allergic reactions. Because the drug can inhibit the satiety center of the hypothalamus and stimulate appetite, it can promote animal growth. . In 2010, the Shanghai Veterinary Drug Feed Testing Institute first discovered that CYP was illegally used by some illegal animal husbandry enterprises as a new type of breeding accelerator, and the residue of CYP in animal products is extremely harmful to the health of consumers. At the end of 2010, the Ministry of Agriculture announced No. 1519 "Substances Prohibited in Feed and Animal Drinking Water", which clearly prohibited the use of such drugs as production accelerators for livestock. CYP monitoring. [0...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/58G01N21/64
CPCG01N21/6408G01N33/533G01N33/582
Inventor 杜霞袁超丁炎
Owner ZHENJIANG YITE BIOTECH DEV CO LTD
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