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A test strip for the differential diagnosis of foot-and-mouth disease virus-infected animals and vaccine-immunized animals

A technology for immunizing animals with foot-and-mouth disease virus is applied in the field of differential diagnosis test strips for animals infected with foot-and-mouth disease virus and vaccine-immunized animals, and can solve the problems of slow release of gold-labeled protein, influence of bacterial protein, and low expression of expressed protein.

Active Publication Date: 2021-07-30
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to overcome the disadvantages of low protein expression in the prior art, difficulty in ensuring natural spatial structure, difficulty in renaturation, bacterial protein affecting the specificity of test results, and the risk of incomplete virus inactivation of inactivated viruses, and existing test strips The existing gold standard protein release is slow and incomplete, which affects the detection accuracy, sensitivity and shelf life of test paper products. The present invention combines the advantages and disadvantages of the existing ELISA method for the detection of foot-and-mouth disease and test strips, and uses artificial antigens prepared by polypeptide epitopes and Gold standard pad treatment method, invented a test strip for the differential diagnosis of foot-and-mouth disease virus-infected animals and vaccine-immunized animals

Method used

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  • A test strip for the differential diagnosis of foot-and-mouth disease virus-infected animals and vaccine-immunized animals
  • A test strip for the differential diagnosis of foot-and-mouth disease virus-infected animals and vaccine-immunized animals
  • A test strip for the differential diagnosis of foot-and-mouth disease virus-infected animals and vaccine-immunized animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Preparation of open-door vacanasovic infection animals and vaccine Immune animal identification diagnostic test strips: first based on foot-and-mouth disease non-structural protein B cell epitope, artificially synthetic foot-and-mouth disease virus non-structural protein 2b, 2c, 3b, 3 epitope polypeptides, and coupled The carrier protein, and the secondary anti-IgG of the antitopent species is prepared, and the detection line (T) and the control line (C) on the cellulose film pad are prepared; the blending pad treatment liquid and the gold bond pad are used. The spray gold label; final assembly strip.

[0033] 1. Synthesis of non-structural protein epitope polypeptides in foot-and-mouth diseased virus:

[0034] Synthetic foot-and-pendulum virus non-structural protein 2b No. 1096 to 1106 amino acids (amino acid sequences are: RTPEDleraerak), 2C, 1414 to 1425, amino acids (amino acid sequences as: HEKVSSSSHPIFKQ), 3B, 1602 to 1613 amino acid sequences (amino acid sequence) Fo...

Embodiment 2

[0056] Foot-and-mouth disease virus infection animals and vaccine Immune Animals Diagnosis Test Banks Preparation As Example 1, the detection line sprayed across the mouthpiece virus non-structural protein 2b No. 1096 to 1106 amino acids (amino acid sequence as: RTPEDleraek, SEQ ID NO.1) ) Polypeptide, in its -NH 4 The end is added to the artificial antigen prepared by cysteine, conjugated carrier protein BSA or OVA.

[0057] Test paper strip detection method:

[0058] (1) Preparation of the sample liquid: Silicide to obtain the blood of the animal, and separated from the serum, taking saline 1: 200 times dilution serum to be tested; the rapid detection of the present invention is inserted into the serum to be detected. The insertion depth does not exceed the marking line, and the test strip is taken after about 30s, and the horizontal is placed from about 1 to 5 minutes, while the results are observed.

[0059] (2) The result is determined: if only one (one) brown red control blo...

Embodiment 3

[0061] Foot-and-mouth disease viral infection animals and vaccine Immune Animals Diagnosis Trial Test Paper Preparation As Example 2, the detection line artificial antigenic polypeptide is a 1414th to 1425 amino acids (amino acid sequence as the non-structural protein 2c) of the foot-and-mouth disease non-structural polypeptide (amino acid sequence) : HEKVSSHPIFKQ, SEQ ID NO.2) Polypeptide.

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Abstract

The invention discloses a test paper strip for differential diagnosis of foot-and-mouth disease virus-infected animals and vaccine-immunized animals. Three epitope polypeptides on viral non-structural proteins 2B, 2C, and 3B. The artificial antigen prepared by the present invention has the advantages of easy acquisition, stable structure, purity up to 99%, low cost, rapid mass production, etc., which solves the problem of low expression of traditionally expressed proteins, difficulty in ensuring natural spatial structure, difficulty in renaturation, difficulty in The removal of bacterial proteins affects the specificity of the test results and other shortcomings, and also solves the risk of incomplete virus inactivation when using inactivated viruses. The invention also pre-treats the gold-labeled pad, which is more conducive to the gold-labeled pad’s water absorption and the release of gold-labeled protein, effectively solving the slow and incomplete release of gold-labeled protein in the existing test paper, which affects the detection accuracy, sensitivity and issues such as shelf life.

Description

Technical field [0001] The present invention relates to a foot-and-mouth disease virus infection animal and vaccine immune animal identification diagnosis test strips, belonging to the field of immunoassay. Background technique [0002] Head-and-mouth disease is a strong infectious diseases that causes hoofed animals caused by foot-and-mouth disease. The foot-and-mouth disease virus has multiple properties, variability, wide range of host, and extremely strong infectivity, so once the pathogenesis is popular, large outbreak. More serious is that foot-and-mouth disease is a strong infectious disease. It is necessary to strictly isolate, block, prohibit animal movement and livestock products, etc., for the pathogenesis, the pathogenesis of the pathogenesis. In order to stop the import and export trade in the onset area and even the onset of livestock products, resulting in huge economic losses. [0003] The main strategy for controlling foot and mouth disease in developing countrie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/09
Inventor 张改平孙亚宁杨苏珍杨继飞陈鑫鑫滕蔓赵东邓瑞广
Owner HENAN ACAD OF AGRI SCI
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