Swine foot-and-mouth disease virus type A antibody enzyme-linked immunosorbent assay kit

A technology of porcine foot-and-mouth disease virus and foot-and-mouth disease virus, which is applied in the direction of virus/bacteriophage, virus, virus peptide, etc., can solve the problems of the difficulty of diagnosis and prevention of foot-and-mouth disease and the inability to strictly distinguish subtypes, and achieve good market prospects, less antigen consumption, The effect of high sensitivity

Active Publication Date: 2020-01-10
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Subtypes cannot be strictly distinguished due to constant antigenic drift
There is almost no cross-immune reaction between serotypes, and there are certain changes in antigenicity between different strains within the same type, which brings great difficulties to the diagnosis and prevention of foot-and-mouth disease

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1, porcine foot-and-mouth disease virus type A antibody ELISA kit coating antigen preparation

[0049] The present invention analyzes the sequence of the foot-and-mouth disease type A strain popular in recent years, uses bioinformatics methods to accurately analyze the antigenic epitope, and screens out the peptide segment suitable for ELISA detection from the main antigenic epitope, that is, foot-and-mouth disease The virus type A antigen epitope polypeptide has a sequence as shown in sequence 1 and sequence 2 in the sequence listing. The polypeptide composition is used as the coating source of the kit of the present invention to prepare a porcine foot-and-mouth disease virus type A antibody ELISA kit.

[0050] 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...

Embodiment 2

[0080] Embodiment 2, composition of porcine foot-and-mouth disease virus antibody ELISA kit

[0081] 1. Evaluation of peptide epitopes

[0082]In order to improve the positive detection rate of the detection method, the screened antigen epitope polypeptide sequence 1 (Seq1), sequence 2 (Seq2) or the mixed peptide (MIX) of the two peptides were combined as the coating antigen (1.0 μg / ml / peptide), according to the conventional indirect ELISA method, 27 porcine foot-and-mouth disease A virus-infected sera were detected. The results showed that the positive detection rates of Sequence 1, Sequence 2 and mixed peptides to 35 infected sera were 81.5%, 92.3% and 100% respectively. The results showed that the mixed peptides could detect all infected sera with a significant positive detection rate. The positive detection rate was higher than that of a single peptide, so the mixed peptide mixed with 2 peptides was used as the coating antigen of the porcine foot-and-mouth disease virus ...

Embodiment 3

[0096] Embodiment 3, the detection method of porcine foot-and-mouth disease virus antibody ELISA kit

[0097] 1.1 Sample preparation Take animal whole blood. After the blood is coagulated, centrifuge at 3500r / min for 10 minutes to collect the supernatant. Blood can also be collected. After coagulation, the serum will naturally precipitate. The serum should be clear and free of hemolysis.

[0098] 1.2 Washing solution preparation Before use, return the 20-fold concentrated washing solution to room temperature. If there is precipitation, dissolve it in a 37°C water bath, and then dilute it 20 times with deionized water for later use.

[0099] 1.3 Sample dilution Dilute the serum to be tested in the serum dilution plate at a ratio of 1:20. The negative and positive control serum has been diluted and can be used directly.

[0100] 1.4 Operation steps

[0101] 1.4.1 Adding samples Take the antigen-coated plate, add the diluted serum to be tested, negative control and positive cont...

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Abstract

The present invention discloses a swine foot-and-mouth disease virus type A antibody enzyme-linked immunosorbent assay kit. The swine foot-and-mouth disease virus type A antibody enzyme-linked immunosorbent assay kit comprises a foot-and-mouth disease virus type A antigen epitope polypeptide coated enzyme-linked reaction plate and an enzyme-labeled anti-antibody; and the foot-and-mouth disease virus type A antigen epitope polypeptide is a polypeptide shown in a sequence 1 or a polypeptide shown in a sequence 2 in a sequence listing. The kit uses the chemically synthesized antigen-peptide-coated reaction plate, is low in antigen consumption and high in sensitivity and specificity, and can efficiently detect presence of foot-and-mouth disease virus infection. The kit has good specificity, issensitive and high in efficiency, and has a good market prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically, the invention relates to an enzyme-linked immunosorbent detection kit for foot-and-mouth disease virus type A antibody, in particular to a pig foot-and-mouth disease virus type A antibody enzyme-linked immunosorbent detection kit. 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 h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09G01N33/569G01N33/543
CPCC07K14/005C12N2770/32122G01N33/54306G01N33/56983G01N2333/09
Inventor 吴冬荀肖进蔡薇郭丽清张蕾
Owner CHINA ANIMAL HUSBANDRY IND
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