Foot-and-mouth disease virus non-structural protein 3B epitope peptide and application thereof

Abstract

The invention discloses a foot-and-mouth disease virus non-structural protein 3B epitope peptide and an application thereof. A hybridoma cell line 2H1 is obtained through screening using a cell fusiontechnology, and a monoclonal antibody secreted by the hybridoma cell line 2H1 has a broad spectrum and a high affinity, and is highly competitive with serum. The antigen epitope recognized by the monoclonal antibody 2H1 is further identified accurately, and the amino acid sequence of the monoclonal antibody 2H1 is represented by SEQ ID No.2; and the antigen epitope peptide is mutated to obtain aseries of mutants. The antigen epitope peptide and the mutants thereof have good reaction ability with FMDV antibody positive serums of foot-and-mouth disease infected pigs, cattle and sheep, and an antibody ELISA detection kit established by using the antigen epitope peptide as envelope antigens can specifically distinguish FMDV infected serum and antigen immunized serum, can be adopted to detectthe serum of any animal species, spans the inter-species barrier, and has the advantages of high reliability, wide application range, good broad spectrum, strong competitiveness, and quickness and convenience in detection.

Description

technical field [0001] The present invention relates to antigenic epitopes of foot-and-mouth disease virus, especially the non-structural protein 3B antigenic epitopes of foot-and-mouth disease virus and their applications. Protein antigenic epitopes and their fields of application. Background technique [0002] Foot-and-mouth disease (FMD) is caused by foot-and-mouth disease virus (FMDV). It is a highly contagious infectious disease that mainly affects livestock such as cattle, pigs, and sheep, as well as a variety of wild cloven-hoofed animals. Humans can also be infected. The disease causes the death of young animals, decreased milk production, decreased meat consumption, decreased meat production, and decreased animal production performance. It is highly contagious and can form a large-scale epidemic, causing huge economic losses. Foot-and-mouth disease virus is a member of the foot-and-mouth disease virus genus of the Picornaviridae family. It is a single-stranded posi...

AI Technical Summary

Problems solved by technology

[0005] (1) The epitope targeted by the monoclonal antibody has not been identified, and the extent of the broad spectrum of the monoclonal antibody cannot be determined: the epitope sequence and its key amino acids or important amino acids targeted by the existing monoclonal antibody are not clear, if If the key amino acids and important amino acids vary between strains, the degree of broad-spectrum will change, which cannot ensure the broad-spectrum of foot-and-mouth disease strains, and correspondingly lacks real application value;

[0006] (2) Poor broad-spectrum of monoclonal antibodies: The antigenic epitopes targeted by monoclonal antibodies are poorly conserved, resulting in no reaction with some strains of foot-and-mouth disease;

[0007] (3) Poor competitiveness of monoclonal antibodies: Some monoclonal antibodies have poor competitive ability with FMD infection serum, which makes it impossible to establish an effective blocking ELISA detection kit

Monoclonal antibodies with no competition or poor competition are only suitable for coating ELISA plates to capture FMDV 3ABC antigen, and then only one of the pig, cow or sheep serum to be tested can be added, and finally HRP is used to label anti-pig, For bovine or sheep IgG antibodies, the differential diagnosis method for foot-and-mouth disease established with this non-competitive or poorly competitive monoclonal antibody is not only cumbersome for warlocks, but more seriously, it cannot cross the barrier between animal species and cannot be used in a reaction system. Serum from any animal species detected in

[0008] Because the existing monoclonal antibodies used to detect foot-and-mouth disease virus have the above-mentioned defects to varying degrees, these monoclonal antibodies cannot establish effective blocking ELISA procedures.

The ELISA detection kits established with these existing monoclonal antibodies, in addition to having low detection sensitivity and poor broad-spectrum (that is, conservation of epitopes), what is more serious is that each animal needs to be detected. Establishing a detection kit lacks practical application value

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