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Time-resolved fluoroimmunoassay kit for detecting adprin as well as detecting method of time-resolved fluoroimmunoassay kit

A time-resolved fluorescence and adefuran technology, which can be used in measurement devices, analytical materials, instruments, etc., can solve the problem of no adefurine, etc., and achieve the effects of simple operation, wide application and strong specificity.

Inactive Publication Date: 2016-02-03
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant research reports on adeffurine in poultry and other animals. In order to complete the declaration of new veterinary drugs, establish the safety evaluation of adeffurine drugs, and ensure food safety, the detection and research of adeffurine is carried out
At present, there are no patents and literature reports on the time-based fluorescent immunoassay for the detection of adefoline

Method used

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  • Time-resolved fluoroimmunoassay kit for detecting adprin as well as detecting method of time-resolved fluoroimmunoassay kit

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Experimental program
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Embodiment

[0024] 1. Preparation of immunogen and coating agent

[0025] Synthesis of the immunogen (adefuroline-BSA) of the present invention: Accurately weigh 324mg of adefuroline and dissolve it in 2mL N,N-dimethylformamide, add γ-aminobutyric acid solution drop by drop under stirring, and stir to react After 3 hours, adjust the pH of the reaction solution to about 10. The precipitate was removed by centrifugation. Add the above reaction dropwise to BSA solution (320mgBSA dissolved in 5mL normal saline), then add 23mg of N-hydroxysuccinimide (NHS), 45.4mg of N,N-dicyclohexylcarbodiimide (DCC), 4 React overnight at ℃, centrifuge to remove the precipitate, take the supernatant and dialyze with phosphate buffered solution (PBS) for 3 days, change the dialysate every 6 hours, freeze-dry the obtained product, and store it at -20 ℃ for future use.

[0026] Synthesis of the original coating (adefuroline-OVA): In the above reaction, after replacing BSA with OVA, the reaction conjugate adefu...

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Abstract

The invention discloses a time-resolved fluoroimmunoassay kit for detecting adprin as well as a detecting method of the time-resolved fluoroimmunoassay kit. The time-resolved fluoroimmunoassay kit consists of a porous coating plate, a buffer solution, an adprin standard product, an adprin antibody lyophilized product, a europium-labeled goat-anti-mouse antibody, a washing solution and an enhancement solution. The detecting method of the time-resolved fluoroimmunoassay kit comprises the following steps: (1) immunogen preparation; (2) dox-ova preparation; (3) monoclonal antibody preparation; (4) sample pretreatment and detection. According to the invention, the detection time is short, the average recovery rate is high, the sample pretreatment is simple, spot operation detection is realized, the application is wide, the detection cost is low, and the advantages that the detection specificity is strong, the intra and inter difference is less, the sensitivity is high, the operation is simple and quick, and the kit is particularly suitable for detection of mass samples are achieved.

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 adfuroline and a detection method thereof. Background technique [0002] Adfurin is a nucleotide compound, which was reported by British scientists in the 1970s. It is a new type of high-efficiency anticoccidial drug being developed in my country. Adefurin has a good therapeutic effect on coccidia, and its anticoccidial index (ACI) is 183.2. Feeding a certain amount of adefurin to Roman male chicks, it can be clearly detected that adefurin exists in the form of the original drug in the chicken liver and kidney. Investigate the acute toxicity test of adefoline with Kunming kind of health, oral administration, after administration once or several times within 24 hours, the toxic reaction and death process produced within a certain period of time can be concluded that adefurin LD 50 3443.5mg?Kg -1 ?bw -1 Af...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/533
Inventor 洪霞薛永来杜霞
Owner JIANGSU WISE SCI & TECH DEV
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