Porcine Foot-and-Mouth Disease Virus Nonstructural Protein 3abc Antibody ELISA Kit

A porcine foot-and-mouth disease virus and non-structural protein technology, applied in the direction of virus/bacteriophage, virus, virus peptide, etc., can solve the problems that it cannot be used as a detection antibody, cannot be used as a detection index, and the repeatability of antibodies is not good, so as to achieve good market prospects and antigenicity Less dosage, high sensitivity and specificity

Active Publication Date: 2018-09-04
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus-infected animals can produce anti-2B antibodies, but this antibody has poor reproducibility in ELISA reactions and cannot be used as a detection antibody
2C antibody fades faster than 3ABC, and 2C antibody can be detected in immunized cattle, so it cannot be used as a detection indicator
Many sources believe that 3ABC is more reliable as a sign of infection

Method used

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  • Porcine Foot-and-Mouth Disease Virus Nonstructural Protein 3abc Antibody ELISA Kit
  • Porcine Foot-and-Mouth Disease Virus Nonstructural Protein 3abc Antibody ELISA Kit
  • Porcine Foot-and-Mouth Disease Virus Nonstructural Protein 3abc Antibody ELISA Kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, porcine foot-and-mouth disease virus nonstructural protein 3ABC antibody ELISA kit coating antigen preparation

[0047] The present invention adopts the bioinformatics method to accurately analyze the epitope of the nonstructural protein, and screens out the peptide segment suitable for ELISA detection from the main epitope on the 3ABC protein, that is, the nonstructural protein 3ABC epitope polypeptide of foot-and-mouth disease virus , and its sequence is shown in sequence 1 in the sequence listing. The polypeptide is used as the coating source of the kit of the present invention to prepare the porcine foot-and-mouth disease virus non-structural protein 3ABC antibody ELISA kit.

[0048] The coated antigen of the present invention can be prepared using an Applied Biosystem automatic polypeptide synthesizer (model 433A). Using the Merrifield solid-phase synthesis method, Fmoc (9-fluorenylmethyloxycarbonyl, 9-fluorenylmethyloxycarbonyl) modified amino acid ...

Embodiment 2

[0079] Embodiment 2, composition of porcine foot-and-mouth disease virus nonstructural protein 3ABC antibody ELISA kit

[0080] Porcine foot-and-mouth disease virus non-structural protein 3ABC antibody ELISA kit includes:

[0081] (1) 96-well detachable polystyrene enzyme-linked reaction plate coated with foot-and-mouth disease virus antigen; 2×96 wells.

[0082] (2) Horseradish peroxidase-labeled rabbit anti-pig IgG polyclonal antibody (purchased from sigma, catalog number A5670), 2 bottles (12 ml each).

[0083] (3) Positive control serum: take the coated antigen artificially synthesized in Example 1 as the immunogen, and immunize the hyperimmune serum prepared from 25-35 day-old specific pathogen-free (SPF) pigs, add 1000U / ml streptavidin and 1000U / ml penicillin, sterilized by 0.2μm filter membrane, used as the positive control serum of the kit. 1 tube (1ml).

[0084] (4) Negative control serum: use normal pig serum without FMD antibody, add 1000U / ml streptomycin and 100...

Embodiment 3

[0091] Embodiment 3, the detection method of porcine foot-and-mouth disease virus nonstructural protein 3ABC antibody ELISA kit

[0092] 1. Equilibration: Take the kit out of the refrigerated environment, put it at room temperature for 30 minutes before use, and mix the liquid reagents before use.

[0093] 2. Dosing: Dilute the concentrated washing liquid 20 times with distilled water or deionized water for later use.

[0094] 3. Setting: leave 1 blank well, 3 negative control wells, and 2 positive control wells, and the rest are sample wells to be tested.

[0095] 4. Pre-dilution of the sample to be tested: add 100 μl of sample diluent to each well of the dilution plate, and add 5 μl of the sample to be tested respectively.

[0096] 5. Adding samples: do not add samples to blank wells; add 100 μl negative control to each of the 3 negative control wells, add 100 μl positive control to each of the 2 positive control wells; add 100 μl diluted samples to be tested in the remaini...

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Abstract

The invention discloses a pig foot-and-mouth disease virus non-structural protein 3ABC antibody enzyme-linked immunosorbent assay kit. The kit comprises an enzyme-linked reaction plate and an enzyme labeling antiantibody, wherein the enzyme-linked reaction plate is coated with foot-and-mouth disease virus non-structural protein 3ABC epitope polypeptide; the foot-and-mouth disease virus non-structural protein 3ABC epitope polypeptide is polypeptide presented by a sequence 1 in a sequence table. According to the kit, the reaction plate coated with non-structural protein 3ABC antigen peptide is chemically synthesized, the consumption of the antigen is low, the sensitivity and the specificity are high, and the foot-and-mouth disease virus infection can be effectively detected. The kit is good in specificity, sensitive and efficient and has good market prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, the invention relates to an ELISA kit for nonstructural protein antibody of foot-and-mouth disease virus, in particular to an ELISA kit for nonstructural protein 3ABC antibody of porcine foot-and-mouth disease virus. Background technique [0002] Foot-and-mouth disease (FMD) is a severe infectious disease caused by foot-and-mouth disease virus (FMDV) infection in cloven-hoofed animals. The fatality rate of foot and mouth disease is very low, but the infection rate is high. The main symptoms of the disease are: blisters in the oral mucosa, tongue, lips, nose mirror, hoof forks, nipples and other parts, ulceration to form rotten spots, sick animals lying on the ground with hoof pain, and severe hoof shell edge cracking or shelling. The symptoms of different animals are slightly different. Pregnant cows may abort, which leads to reduced fertility; pigs have broken hooves as the main ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/09G01N33/68G01N33/569
CPCC07K14/005C12N2770/32122G01N33/56983G01N33/6803G01N2333/09
Inventor 齐鹏董春娜王楠张蕾肖进赵洪涛张君张艳宾张欣
Owner CHINA ANIMAL HUSBANDRY IND
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