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Foot and mouth disease virus liquid phase blocking BSA-ELISA antibody detection kit and application

A foot-and-mouth disease virus and antibody detection technology, applied in the biological field, can solve the problems of operation fatigue, cumbersome operation, error-prone, etc., and achieve the effects of improving efficiency, improving economic benefits, and improving stability

Pending Publication Date: 2021-07-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The BSA-ELISA antibody detection kit shortens the operation of the liquid-phase blocking ELISA method and easily completes it within half a working day, solving the problems of cumbersome, long-time-consuming, fatigue-inducing, unstable, and error-prone operations

Method used

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  • Foot and mouth disease virus liquid phase blocking BSA-ELISA antibody detection kit and application
  • Foot and mouth disease virus liquid phase blocking BSA-ELISA antibody detection kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 foot-and-mouth disease virus type O liquid phase blocking BSA-ELISA kit development and application

[0032] 1 Materials and methods

[0033] 1.1 Preparation of foot-and-mouth disease virus antigen and antiserum

[0034] 1.1.1 Concentration and freeze-drying of the FMDV antigen required by the kit: add an appropriate volume of inactivated foot-and-mouth disease virus vaccine (O / Mya98 strain) to 8% PEG6000, 4% NaCl, stir at 4°C for 4 hours, and then stand at 4-8°C Overnight, centrifuge the 500ml centrifuge tube at 6000rmp for 60 minutes, discard the supernatant, continue to add uncentrifuged antigen for centrifugation, repeat the above operation when the amount of antigen is large, and finally suspend the antigen precipitate in the centrifuge tube with pH 7.6 0.01mol PBS, and dilute to the original antigen 1 / 50 of the volume, add an equal volume of cold trichlorethylene to emulsify and centrifuge to degrease, absorb the antigen layer liquid, and use the liq...

Embodiment 2

[0110] Embodiment 2 Foot-and-mouth disease virus type A liquid phase blocking BSA-ELISA kit development and application

[0111] 1 Materials and methods

[0112] 1.1 Preparation of foot-and-mouth disease virus antigen and antiserum

[0113] 1.1.1 Concentration and freeze-drying of the FMDV antigen required by the kit: add an appropriate volume of inactivated foot-and-mouth disease virus vaccine (AF72 strain) to 8% PEG6000, 4% NaCl, stir at 4°C for 4 hours, and then stand at 4-8°C overnight. Centrifuge the 500ml centrifuge tube at 6000rmp for 60 minutes, discard the supernatant, continue to add uncentrifuged antigen for centrifugation, repeat the above operation when the amount of antigen is large, and finally suspend the antigen precipitate in the centrifuge tube with 0.01mol PBS of pH 7.6, and dilute to the volume of the original antigen. 1 / 50, add an equal volume of cold trichlorethylene to emulsify and centrifuge to degrease, absorb the antigen layer liquid, use the liquid...

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Abstract

The invention relates to a foot-and-mouth disease virus liquid-phase blocking BSA-ELISA antibody detection kit and application, and belongs to the technical field of biologication. The foot-and-mouth disease virus liquid-phase blocking BSA-ELISA antibody detection kit is characterized in that a foot-and-mouth disease virus antigen is treated through a concentration and freeze-drying process; a novel enzyme-linked immunosorbent assay method is established by combining guinea pig anti-IgG-biotin and HRP-SA, and meanwhile, the operation mode is improved. The problems that a foot-and-mouth disease virus liquid phase blocking ELISA antibody detection kit is tedious in operation, multiple in steps, long in time consumption, poor in stability and prone to making mistakes, a pig serum sample is prone to inversion, and the production batch is small are solved. The kit is convenient to operate, time is shortened, batch production is achieved, the sensitivity and specificity of a classic method are kept, the stability of transportation and storage is improved, the validity period of the kit is prolonged, and technical support is provided for the method used for foot-and-mouth disease immune antibody monitoring, vaccine immune effect evaluation, epidemiological investigation, immune guidance and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a liquid-phase blocking BSA-ELISA antibody detection kit for foot-and-mouth disease virus and its application. Background technique [0002] Foot and Mouth Disease (FMD) is an acute, febrile, highly contagious infectious disease that is susceptible to cloven-hoofed animals such as pigs, cattle, and sheep caused by Foot and Mouth Disease Virus (FMDV). Most of the clinical features are vesicular eruptions on the oral mucosa, hooves and breast skin. The disease spreads through many routes and is fast. Every outbreak will cause huge economic losses to the local animal husbandry industry. The World Organization for Animal Health (OIE) lists it as an infectious disease that must be declared, and my country also lists it as the first class of animal infectious diseases. Foot-and-mouth disease virus mainly has seven serotypes in the world: O, A, Asia 1, C and South Africa 1, 2, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/58G01N33/543
CPCG01N33/56983G01N33/581G01N33/54306G01N2333/09G01N2469/20
Inventor 周广青马军武邵军军常惠芸龚真莉祁淑芸马米灵李苗苗何继军郭建宏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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