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Virus-like particles based type-A foot and mouth disease virus antibody detection kit

A technology of foot-and-mouth disease virus and detection kit, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor immunogenicity of a single protein or polypeptide, affect the effect of use, and many experimental steps, so as to achieve good immunogenicity, Good safety, high safety effect

Inactive Publication Date: 2019-05-24
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

[0003] Most of the ELISA methods currently used to detect FMDV antibodies use inactivated viruses, recombinant viruses, single proteins or polypeptides as antigens. Although using intact virus particles as antigens has many advantages, it has safety risks. Immunogenicity is relatively poor, affecting the effect of its use
[0004] At present, most competitive ELISA kits mark HRP on the anti-competing antibody antibody, which leads to many experimental steps and takes a long time, mostly more than 1 h. content

Method used

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Examples

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Embodiment Construction

[0017] The present invention is illustrated below in conjunction with embodiment.

[0018] 1. Preparation of Type A Foot-and-Mouth Disease Virus VLPs

[0019] (1) Co-transform the positive recombinant plasmids pSMK-VPOVP3 and pSMK-VP1 preserved in our laboratory into competent cells BL21(DE3)-RIL, and inoculate the expressing bacteria with 10 μg / mL kanamycin at a ratio of 1:100. , 50 μg / mL ampicillin and 25 μg / mL chloramphenicol in 100 mL of fresh LB liquid medium sterilized by autoclaving at 37 ° C and 220 r / min on a shaker overnight.

[0020] (2) Inoculate the overnight cultured expression bacteria into 1000mL fresh LB liquid medium that was sterilized by autoclaving containing 1 μg / mL kanamycin, 50 μg / mL ampicillin and 25 μg / mL chloramphenicol. Shake on a shaker at 220r / min until the OD600 value of the bacterial solution is 0.6~0.8.

[0021] (3) Add IPTG with a final concentration of 0.5mmol / L, and induce at 16°C for 16h.

[0022] (4) Collect bacteria

[0023] (5) Sonic...

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PUM

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Abstract

The invention discloses a virus-like particles based type-A foot and mouth disease virus antibody detection kit and a preparation method thereof. The virus-like particles based type-A foot and mouth disease virus antibody detection kit comprises an elisa plate coating the virus-like particles, HRP labeled rabbit antibody, a sample diluent containing phosphate buffer, a cleaning solution containingTween, a TMB substrate, positive control serum, negative control serum, and stop buffer mixed of concentrated sulfuric acid and water. Compared with the prior art, the kit has better safety, good immunogenicity and higher flexibility and specificity.

Description

technical field [0001] The invention relates to an antibody detection kit and a preparation method thereof. Specifically, the invention relates to a detection kit prepared by using type A foot-and-mouth disease virus-like particles (VLPs) and a preparation method thereof. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infectious disease caused by foot-and-mouth disease virus (FMDV), which mainly infects cloven-hoofed animals such as pigs, cattle, and sheep. FMDV has 7 serotypes: O, A, C, SAT1, SAT2, SAT3, and Asia I, and there is almost no cross-immune protective reaction among these 7 serotypes. Due to the extremely fast spread of FMD, the constant changes in serotypes and genotypes, and the lack of cross-protection between serotypes, it has brought great challenges to the prevention and control of FMD. In recent years, types A and O have appeared alternately in some areas of China, and the epidemics are complex, involvin...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543G01N33/535
Inventor 郭慧琛孙世琪白满元张韵张禹茹嘉喜杨志元王蕊何继军罗建勋
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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