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A multi-antigen ELISA kit for detecting antibodies against African swine fever virus

An African swine fever virus and kit technology, which is applied in the fields of biotechnology and animal infectious disease diagnosis and research, can solve the problems of high price, difficult preparation of recombinant antigens, and limited supply of kits.

Inactive Publication Date: 2015-09-02
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, ELISA is an international trade inspection method stipulated by OIE, but the detection antigen needs to be prepared from virus-infected cells, virus culture has the risk of spreading the virus, and the diagnostic reliability needs to be improved. The results need to be verified by methods such as Western-blotting ;In order to solve these problems, recombinant antigen ELISA attempts are being carried out at home and abroad. Commercial kits have been supplied in France and Spain. The test results of strains need to be further verified

Method used

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  • A multi-antigen ELISA kit for detecting antibodies against African swine fever virus
  • A multi-antigen ELISA kit for detecting antibodies against African swine fever virus
  • A multi-antigen ELISA kit for detecting antibodies against African swine fever virus

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Embodiment Construction

[0037] Source of biological material:

[0038] pGEX-4T-1-B602L: containing the E70 ASFV B602L gene, donated by Dr. RM Parkhouse of the Gulbenkian de Cie^ncia Institute in Portugal (Reis, A.L., R.M.Parkhouse, A.R.Penedos, C.Martins, and A.Leitao.2007. Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus. J.Gen.Virol.88:2426-2434.);

[0039] pGEX-4T-1-E183L: Contains the E183L (p54) gene of E70 strain ASFV, donated by Dr. RM Parkhouse of the Gulbenkian de Cie^ncia Institute in Portugal (Reis, A.L., R.M.Parkhouse, A.R.Penedos, C.Martins, and A.Leitao .2007.Systematic analysis of longitudinal serological responses of pigs infected experimentally with African swine fever virus.J.Gen.Virol.88:2426-2434.);

[0040] pGEX-4T-1-K205R: containing the E70 strain ASFV K205R gene, donated by Dr. RM Parkhouse of the Gulbenkian de Cie^ncia Institute in Portugal (Reis, A.L., R.M.Parkhouse, A.R.Penedos, C.Martins, and A.Leitao...

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Abstract

The invention provides a multi-antigen enzyme linked immunosorbent assay (ELISA) kit for detecting an African swine fever virus (ASFV) antibody, belonging to the field of a biotechnology and diagnosis and research of animal-borne diseases. The multi-antigen enzyme linked immunosorbent assay kit comprises expression and purification of three types of ASFV recombined antigens, preparation of positive and negative control blood serum of an ASFV antibody, optimal envelope antigen combination and concentration determination, optimization of multi-antigen ELISA (MA-ELISA (Microalbumin-Enzyme Linked Immunosorbent Assay)) reaction parameters, determination of an ASFV antibody negative blood serum critical value, MA-ELISA detection artificial infection and determination of sensitivity, specificity and repeatability of field blood serum samples. By detecting and testifying a large quantity of known blood serum samples, the sensitivity, the specificity and the repeatability of detecting the ASFV antibody by the MA-ELISA are obviously higher than those of an ELISA method recommended by World Organization for Animal Health and oversea similar kits; and the multi-antigen enzyme linked immunosorbent assay kit can be used for ASFV serological diagnosis, epidemiological investigation and live pig import and export quarantine inspection.

Description

technical field [0001] The invention relates to the fields of biotechnology and diagnosis and research of animal infectious diseases. Background technique [0002] African swine fever (ASF) is a highly fatal infectious disease of pigs. The disease has been prevalent in many African countries and has now spread to neighboring countries such as Georgia, Armenia, Azerbaijan and Russia. The threat is growing. At present, there is no vaccine for ASF, so rapid and accurate diagnosis is very important to prevent the introduction of the disease and its control. Diagnostic techniques for ASF recently recommended by the World Organization for Animal Health (OIE) include pathogen identification and serological tests. [0003] In pathogen identification technology, hemadsorption test is one of the classic ASF identification methods with high sensitivity, but it needs temporary preparation and culture of porcine bone marrow cells, which is not only time-consuming and cumbersome to oper...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569
Inventor 张鑫宇孙怀昌夏晓莉
Owner YANGZHOU UNIV
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