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A-type foot-and-mouth disease competition ELISA antibody detection kit

A technology for antibody detection and foot-and-mouth disease, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to quantitatively evaluate antibody titers, cumbersome operation steps, and large human errors, so as to save raw materials, accurate and sensitive detection, and promote The effect of accuracy

Inactive Publication Date: 2015-07-22
LUOYANG LAIPSON INFORMATION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The liquid-phase blocking method can not only detect virus infection but also detect antibodies to evaluate the immune effect of vaccines. It has been widely promoted in the market, but it also has disadvantages such as time-consuming, cumbersome operation steps, and large human errors; although the indirect method is simple to operate , is faster, but cannot quantitatively evaluate the antibody titer in the sample

Method used

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  • A-type foot-and-mouth disease competition ELISA antibody detection kit
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preparation example Construction

[0032] The preparation method of the type A foot-and-mouth disease rabbit antiserum comprises the following steps:

[0033] 1) Mix and emulsify type A foot-and-mouth disease recombinant protein with complete Freund's adjuvant in a volume ratio of 1:1 to prepare an emulsion for later use;

[0034] 2) First immunize rabbits with a weight of 1.8-2.2kg, and boost the rabbits with emulsion every two weeks;

[0035] 3) On the 7th day after the last booster immunization, blood was collected from the ear vein to detect the serum antibody titer. When the OD450nm value reached 1.9-2.1, blood was collected from the heart, and the serum was purified to obtain the antiserum for type A foot-and-mouth disease rabbits.

[0036] The detection method of described type A foot-and-mouth disease competition ELISA antibody detection kit may further comprise the steps:

[0037] 1. Carry out sample layout on the 96-well reaction plate, which is divided into sample column, positive control column, ...

Embodiment 1

[0044] A type A foot-and-mouth disease competition ELISA antibody detection kit, said detection kit includes washing solution, sample diluent, chromogenic solution, stop solution, and also includes:

[0045] (1) Reaction plate coated with type A foot-and-mouth disease polyantibody: use type A foot-and-mouth disease rabbit antiserum as the coating antibody, dilute it with PB buffer to 1:4000, and add 100ul / well to the 96-well reaction plate, Coat at 4°C for 20 hours, wash with washing solution 2-3 times, add blocking solution 200ul / well, seal at 4°C for 16 hours, then shake off liquid, dry at 37°C for 3 hours, pack into bags and seal.

[0046] (2) Type A foot-and-mouth disease enzyme-labeled monoclonal antibody: Horseradish peroxidase-labeled type A foot-and-mouth disease monoclonal antibody.

[0047] (3) Type A foot-and-mouth disease virus antigen: used for quantitative detection of antibody in samples.

[0048] (4) Negative and positive control serum for type A foot-and-mout...

Embodiment 2

[0070] Preparation of a type A foot-and-mouth disease competition ELISA antibody detection kit:

[0071] 1. Mix and emulsify type A foot-and-mouth disease recombinant protein with Freund's complete adjuvant 1:1, and then select about 2 kg of rabbits for the first immunization; use type A foot-and-mouth disease recombinant protein and Freund's incomplete adjuvant to carry out 1:1 mixed emulsification Booster immunization every two weeks; 7 days after the last booster, blood was collected from the ear vein to detect the serum antibody titer. When the titer reached the requirement, blood was collected from the heart, and the serum was purified to prepare type A Foot-and-mouth disease rabbit antiserum.

[0072] 2. Preparation of the reaction plate coated with polyantibody against type A foot-and-mouth disease: use rabbit antiserum against type A foot-and-mouth disease as the coating antibody

[0073] Diluted with PB buffer to 1:4000, added 100ul / well to a 96-well reaction plate, ...

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Abstract

The invention belongs to the field of biological detection and particularly relates to an A-type foot-and-mouth disease competition ELISA antibody detection kit. By adopting the kit to improve the original in-vitro reaction process, the reaction time is greatly shortened from the original 4 h to 1.5 h, the original accessories of the in-vitro reaction are saved, and the kit is more rapid and economic. The kit adopts an enzyme-labeled antibody to carry out reaction, so that compared with the traditional enzyme-labeled secondary antibody process, the detection is more correct and sensitive. The reaction plate coated by using the method provided by the invention is strong in adsorbability and is capable of effectively promoting the correctness of the detection.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to a type A foot-and-mouth disease competition ELISA antibody detection kit. Background technique [0002] Foot-and-mouth disease is a highly contagious cloven-hoofed animal disease caused by foot-and-mouth disease virus. Due to its strong transmission and high incidence, it seriously affects the development of animal husbandry in various countries. At the same time, it is also a zoonotic disease, which is extremely harmful to society. Has been classified as a class of infectious diseases by the Ministry of Agriculture. According to the serotypes of foot-and-mouth disease, it is divided into seven types: A, O, C, AsiaⅠ, SATⅠ, SATⅡ, and SATⅢ. Each type has several subtypes, and the antigenicity between them is very different. Work brings many difficulties. It is necessary to develop a practical, sensitive and specific detection and diagnosis method. [0003] At present, the dia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 王善普刘玉宝杜小波张鹏翼智雪玲李权伟李晓杨少杰董琴
Owner LUOYANG LAIPSON INFORMATION TECH
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