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Specific antigen polypeptide for reticuloendotheliosis virus of poultry and application

A poultry reticuloendothelial and specific technology, applied in the direction of viral peptides, viruses, specific peptides, etc., can solve the problems of large differences between antigen batches, differences, and difficulty in prevention and control, and achieve convenient operation, high antigen purity, preparation and sample Quick and easy results of detection

Inactive Publication Date: 2019-04-16
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Avian endothelial histoproliferation virus (REV) is a type C retrovirus of poultry in the Retroviridae family, which can cause acute and chronic tumor formation, immunosuppression, dwarf syndrome, etc., and bring huge economic losses to poultry production
Due to the frequent mixed infection of the disease, the prevention and control of the disease becomes very difficult
Currently, the methods used to detect REV antibodies include IFA, ELISA, etc. However, there are often differences in the results of different kits and different methods, mainly due to the differences in the antigens used.
The commercialized ELISA kits commonly used in the past used the purified virus to be properly processed for antigen coating. Some scholars also used fusion expression proteins as the coating antigen. However, these methods often caused large batch-to-batch differences or non-specific issues

Method used

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  • Specific antigen polypeptide for reticuloendotheliosis virus of poultry and application
  • Specific antigen polypeptide for reticuloendotheliosis virus of poultry and application
  • Specific antigen polypeptide for reticuloendotheliosis virus of poultry and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Use biological software to analyze the amino acid sequence of REV gp90, select the amino acid sequence No.5 with better antigenicity, and determine several possible antigenic fragments from the hydrophilicity, antigenic index, surface site index, and gp90 conservation , and the amino acid sequences are respectively No.1, No.2, No.3, No.4, and No.5. Polypeptide antigen synthesis was carried out according to conventional methods, and the enzyme plate was coated with polypeptide antigen, and REV positive serum and SPF chicken serum at different infection stages were detected, and the results were shown in Table 1 (values ​​are OD values):

[0024] No.1: QKTKLGSYEDGPNKL

[0025] No. 2: GGGPTDMIREESVRERLEEII

[0026] No. 3: LPRPRGVDLDPQTSDILEA

[0027] No.4: SGEEPIPLPSIEYTAGRHK

[0028] No.5: GVDLDPQTSDILEATHQVPSVQYHPLALPRP

[0029] Table 1 Reaction of No.1-5 polypeptides with REV positive serum and negative serum

[0030]

[0031] Clearly, there are significant diff...

Embodiment 2

[0033] Composition of the kit

[0034] The composition of the kit in this example is as follows:

[0035] [1] A microtiter plate coated with polypeptide antigen;

[0036] [2] Enzyme-labeled antibody: horseradish peroxidase-labeled rabbit anti-chicken IgG;

[0037] [3] Substrate solution: commercial TMB substrate solution;

[0038] [4] Stop solution 1% SDS: take 100.0mL of distilled water and 1.0g of SDS powder and mix them;

[0039] [5] Washing solution: add 0.5mL Tween-20 to 1000mL10mmo1 / L PBS with pH7.4;

[0040] [6] Negative control and positive control.

Embodiment 3

[0042] Polypeptide antigen No.5 was coated with or without antigen coupling, and the serum of REV at different infection stages was tested. The test results are shown in Table 2 (values ​​are OD values):

[0043] Table 2 The detection results of polypeptide antigen No.5 coupled or uncoupled antigen coating

[0044]

[0045]

[0046] It can be seen from the above test results that although there are some differences between the naked peptide antigen and the conjugated polypeptide antigen in the detection, the difference is not significant. Therefore, both conjugated and unconjugated polypeptide antigens have good detection results.

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Abstract

The invention relates to a specific antigen polypeptide for a reticuloendotheliosis virus of poultry and application of the polypeptide. The amino acid sequence of the specific antigen polypeptide isSEQ ID No.5; the specific antigen polypeptide is coded by envelope protein gp90 gene of the virus, and has strong reactogenicity, the epitope of the antigen has good antibody reactivity, and can be used for specifically detecting the antibody of the REV, and the specific antigen polypeptide has the application value of clinical detection. The antibody for specifically resisting the REV can be prepared by utilizing a polypeptide antigen immune animal.

Description

technical field [0001] The invention relates to an avian reticuloendothelial disease virus-specific antigen polypeptide and its application. Background technique [0002] Avian endothelial disease virus (REV) is a type C retrovirus of poultry in the Retroviridae family, which can cause acute and chronic tumor formation, immunosuppression, dwarf syndrome, etc., and bring huge economic losses to poultry production. Because the disease often occurs mixed infection, the prevention and control of the disease becomes very difficult. Currently, the methods used to detect REV antibodies include IFA, ELISA, etc. However, the results of different kits and different methods often show differences, which is mainly due to the differences in the antigens used. The commercialized ELISA kits commonly used in the past used the purified virus to be properly processed for antigen coating. Some scholars also used fusion expression proteins as the coating antigen. However, these methods often c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/15C07K16/10G01N33/68G01N33/569
CPCC07K14/005C07K16/1036C12N2740/13022G01N33/56983G01N33/6854G01N2333/15G01N2469/10G01N2469/20
Inventor 秦爱建王娅娟杨叶钱琨邵红霞叶建强
Owner YANGZHOU UNIV
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