Time resolution fluorescence immunoassay kit for detecting aflatoxin M1

A time-resolved fluorescence and aflatoxin technology, applied in fluorescence/phosphorescence, measurement devices, analysis materials, etc., to achieve simple pre-treatment, wide application, and strong specificity

Inactive Publication Date: 2018-06-29
JIANGSU WISE SCI & TECH DEV
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  • Abstract
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  • Application Information

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Problems solved by technology

There are no patents and literature reports on the chronofluorescence immunoassay for the detection of aflatoxin M1

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  • Time resolution fluorescence immunoassay kit for detecting aflatoxin M1

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Embodiment

[0024] 1. Preparation of immunogen and coating agent

[0025] Synthesis of the immunogen (aflatoxin M1-BSA) of the present invention: Accurately weigh 324 mg of aflatoxin M1 and dissolve in 2mL N,N-dimethylformamide, add γ-aminobutyric acid solution drop by drop 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;

[0026] Synthesis of the original coating (aflatoxin M1-OVA): In the above reaction, after replacing BSA with OVA, the reaction conjuga...

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Abstract

The invention discloses a time resolution fluorescence immunoassay kit for detecting aflatoxin M1. The time resolution fluorescence immunoassay kit for detecting aflatoxin M1 consists of a porous coating plate, a buffer solution, an aflatoxin M1 standard substance, an aflatoxin M1 antibody freeze-drying product, a europium-labeled goat anti-rat antibody, a scrubbing solution and an enhancing solution. A detection method of the time resolution fluorescence immunoassay kit for detecting aflatoxin M1 comprises the following steps: (1) preparation of an immunogen; (2) preparation of a coating source; (3) preparation of a monoclonal antibody; and (4) pretreatment and detection of a sample. The time resolution fluorescence immunoassay kit is short in detection time, high in average recovery rate, simple in sample pretreatment, wide in application, low in detection cost, high in detection specificity, small in intra-batch and batch-to-batch difference, high in sensitivity and simple and quickin operation, is capable of field operation detection, and is particularly applicable for the detection of large-scale samples.

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 aflatoxin M1. Background technique [0002] Aflatoxin M1 (Aflatoxin M1, abbreviated as AFM1) belongs to a class of compounds with similar structures to aflatoxins. This toxoid is a metabolite produced by common Aspergillus flavus and Aspergillus parasiticus. Feeds have the highest incidence of aflatoxins. [0003] AFM 1 It is a metabolite of aflatoxin M1, and its basic structure is a difuran ring oxinone. Soluble in many organic solvents, such as chloroform, acetonitrile, methanol and water, but insoluble in non-polar solvents such as n-hexane, petroleum ether, ether, etc. The physical and chemical properties are quite stable and will not be destroyed by pasteurization. [0004] Aflatoxin is a strong carcinogen and a highly toxic agent of fungi such as Aspergillus. Most government agencies have strict re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/577
CPCG01N21/6408G01N21/6486G01N33/577G01N2333/38
Inventor 杜道林薛永来洪霞
Owner JIANGSU WISE SCI & TECH DEV
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