Kit for detecting South Africa II-type foot-and-mouth disease virus antibody and preparation method

The technology of a foot-and-mouth disease virus and detection kit is applied in the field of South African type 2 foot-and-mouth disease virus antibody competition ELISA detection kit, which can solve the problems of time-consuming and many experimental steps, and achieve the effects of strong immunogenicity, simple experiment and cost reduction

Inactive Publication Date: 2019-05-24
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, most competitive ELISA kits mark HRP on the anti-competing antibody antibod

Method used

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Examples

Experimental program
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Example Embodiment

[0016] The present invention is explained below with examples.

[0017] 1. Preparation of South African Foot-and-Mouth Disease Virus Type 2 VLPs

[0018] (1) The South Africa type 2 foot-and-mouth disease virus (refer to the AJ251473 strain in GenBank) positive recombinant plasmids pSMK-VP0VP3 and pSMK-VP1 stored in our laboratory were co-transformed into competent cells BL21(DE3)-RIL, and the monoclonal colonies were picked Inoculate at a ratio of 1:100 into a freshly autoclaved LB liquid medium containing 10μg / mL kanamycin, 50μg / mL ampicillin and 25μg / mL chloramphenicol, shake at 37℃, 220r / min Incubate overnight in bed. VP0, VP1, and VP3 and the gene sequence are SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3, respectively.

[0019] (2) Inoculate the expression bacteria cultured overnight into the LB medium containing the above 3 kinds of antibiotics, and cultivate them at 37°C at 220 r / min until the OD600 value is about 0.6-0.7. Reserve 1 ml of the sample as pre-induction Contras...

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Abstract

The invention discloses a detection kit prepared by adopting South Africa II-type foot-and-mouth disease virus-like particles (VLPs) and a preparation method. The preparation method includes: directlymarking HRP on a competitive antibody acting at the same time with a detected antibody, wherein South Africa II-type foot-and-mouth disease virus-like particles coating an endoenzyme target are formed by assembling structural protein VP0, VP1 and VP3 of South Africa II-type foot-and-mouth disease. The VLPs can be assembled efficiently without having impact on their diagnosis efficiency and are conducive to lowering cost of a diagnosis preparation. Compared with the prior art, the method has the advantages that detection steps are reduced, experiment operation is simple, workload is low, consumed time is short, and specificity and sensitivity of a diagnosis reagent can be improved.

Description

technical field [0001] The invention relates to a South African type 2 foot-and-mouth disease virus antibody competition ELISA detection kit. Background technique [0002] Foot-and-mouth disease occurs in Asia, Africa, the Middle East, and South America, causing huge economic losses. The World Health Organization lists foot-and-mouth disease as a legally notifiable infectious disease. South African Foot-and-Mouth Disease Virus Type 2 (SAT2) is a member of the genus Foot-and-Mouth Disease Virus and Picornaviridae. In recent decades, reports of South African type 2 foot-and-mouth disease virus have appeared in the surrounding areas outside the African continent, which shows that the virus has appeared in the danger of cross-border transmission. In 2000, there were SAT2 foot-and-mouth disease epidemics in Kuwait and Saudi Arabia. In 2012, Pakistan There have also been cases of SAT2 foot-and-mouth disease in India and Bahrain. It can be seen that SAT2-FMDV is threatening all c...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543G01N33/535
Inventor 郭慧琛张韵孙世琪董虎白满元侯凤萍茹嘉喜
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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