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Indirect ELISA kit for detecting African swine fever virus

An African swine fever virus and kit technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as loss of activated apoptotic protease

Inactive Publication Date: 2011-11-09
陈文刚
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mutant of P54 lacks 13aa and loses the function of activating apoptosis protease (Alejo A, GAndrés, ML Salas. African Swine Fever Virus Proteinase Is Essential for Core Maturation and Infectivity Journal of Virology, 2003, 77: 5571~557)

Method used

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Embodiment Construction

[0024] Below in conjunction with specific embodiment the present invention is described in further detail:

[0025] Technical scheme of the present invention is as follows:

[0026] 1. Antigen preparation

[0027] 1. Expression of African swine fever P54 protein

[0028] According to the African swine fever (ASFV) P54 gene sequence in GenBank, a pair of primers were designed and synthesized, and the P54 gene fragment was amplified from the ASFV DNA by PCR method, which was cloned into the expression vector pET28b to construct the recombinant plasmid pET-P54 After sequencing verification, it was transformed into the expression host strain BL21(DE3), and its expression was induced by IPTG.

[0029] The designed primers are:

[0030] P54-1-5'-GC GGATCC CATTATTTATCATCGTT-3'

[0031] P54-2-5'-AG CTCGAG CAAGGAGTTTTTCTA-3'

[0032] The underlined part is the restriction site introduced, P54-1 is the upstream amplification primer of ASFV P54 gene, the restriction site introduc...

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Abstract

The invention discloses an indirect ELISA kit for detecting African swine fever virus (ASFV) antibody, and a use thereof, and belongs to the technical field of biology. The kit is characterized by: adopting prokaryotic expressed recombined P54 protein as coating antigen; detecting antibody against to the ASFV in swine serum according to an indirect ELISA principle. The coating antigen in 96-well plates of the kit is the prokaryotic expressed recombined P54 protein which has good antigenicity. The enzyme-linked immunospecific assay kit provided by the present invention comprises the 96-well plates coated by the P54 protein, positive control, negative control, horseradish peroxidase-labeled rabbit anti-porcine IgG polyclonal antibody, a concentrated cleaning solution, a serum dilution, a TMB substrate indicator and a stop buffer. The kit provided by the present invention can be provided for screening mass samples. In addition, the main reagents in the kit are provided in a working fluid manner such that the reagents are used conveniently.

Description

technical field [0001] The invention is a detection kit for detecting African swine fever virus (ASFV) antibody, and belongs to the field of biotechnology. Specifically, the present invention relates to an ELISA kit for detecting antibodies in animal quarantine and its application. Background technique [0002] African swine fever (ASF) is an acute, febrile, highly contagious infectious disease of pigs caused by African swine fever virus (ASFV). It is characterized by a short course of disease and a high fatality rate, which can be as high as 100%. The clinical symptoms and pathological changes are similar to acute swine fever, and it is easy to be misdiagnosed during diagnosis. (William, Hess Adv.African swine fever: a reassessment[J].Vet Sci Comp Med, 1981, 25:39~69) The World Organization for Animals (OIE) listed it as a class A disease, and our country stipulates that it is a class I animal disease, which is subject to All countries in the world attach great importance...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/535
Inventor 陈文刚董志珍王涛
Owner 陈文刚
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