Time resolution fluorescence immunoassay kit for detecting aflatoxins M1

A time-resolved fluorescence and aflatoxin technology, applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of long analysis time, high test cost, cumbersome pretreatment steps, etc., and achieve simple pretreatment, wide application and easy operation easy effect

Inactive Publication Date: 2019-05-14
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The chemical detection method has the disadvantages of high test cost, long analysis time, large sample volume (usually >500 mL) and cumbersome pretreatment steps, etc., and cannot systematically, comprehensively and quickly analyze the content of aflatoxin in a wide range
There are no patents and literature reports on the chronofluorescence immunoassay for the detection of aflatoxin M1

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0022] 1. Preparation of immunogen and coating agent

[0023] 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;

[0024] Synthesis of the original coating (aflatoxin M1-OVA): In the above reaction, after replacing BSA with OVA, the reaction conjuga...

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PUM

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Abstract

The invention discloses a time and fluorescence resolution fluorescence immunoassay kit for detecting aflatoxins M1. The time resolution fluorescence immunoassay kit for detecting the aflatoxins M1 iscomposed of a porous coating plate, buffer liquid, an aflatoxins M1 standard substance, an aflatoxins M1 antibody freeze-drying substance, a europium-labeled goat anti-mouse antibody, a cleaning solution and an enhancement solution. A detecting method of the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1 comprises the following steps that (1) preparation of immunogen; (2) preparation of coating antigen; (3) preparation of monoclonal antibody; and (4) pretreatment and detection of a sample. According to the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1, the detecting time is short, the average recovery rate is high, the pretreatment of the sample is simple, site operation detection can be achieved, the application is wide, and the detection cost is low; and the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1 synchronously has the advantages that the detecting specificity is high, the intra-batch and inter-batch differences are small, the sensitivity is high and the operation is simple and rapid, the time and fluorescence resolution fluorescence immunoassay kit for detecting the aflatoxins M1 is particularly suitable for detection of large scale of 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 aflatoxin M1. Background technique [0002] Aflatoxins (AFT) are secondary metabolites of fungi, mainly produced by some strains of Aspergillus flavus, A. parasiticus and A. nonius; The toxin-producing ability of Aspergillus parasiticus varies greatly from strain to strain; all strains of Aspergillus parasiticus can produce aflatoxin. Aflatoxin is a highly toxic hepatotoxin that can cause acute or chronic damage to the liver. It and hepatitis B virus are considered to be the two main causes of liver cancer; A variety of other tissues and organs can also cause serious damage. What is more serious is that aflatoxin has been proven to have the "three causes" of carcinogenicity, teratogenicity, and cell mutation. Aflatoxins are widely found in grain agricultural products such as peanuts, corn, wheat, and rice,...

Claims

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Application Information

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