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Competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) kit for detecting antibody of African swine fever virus and application thereof

An African swine fever virus and kit technology, which is applied to an ELISA kit for detecting antibodies in animal quarantine. For the detection of African swine fever virus antibody detection kits, it can solve problems such as loss of activated caspase and achieve safety. Good, reliable effect, stable production effect

Inactive Publication Date: 2010-09-15
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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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 VirusProteinase Is Essential for Core Maturation and Infectivity Journal of Virology, 2003, 77: 5571~557)

Method used

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  • Competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) kit for detecting antibody of African swine fever virus and application thereof
  • Competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) kit for detecting antibody of African swine fever virus and application thereof
  • Competitive ELISA (Enzyme-Linked Immuno Sorbent Assay) kit for detecting antibody of African swine fever virus and application thereof

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Embodiment Construction

[0024] The hybridoma cell line of the monoclonal antibody against African swine fever virus is deposited in CGMCC NO.3750, General Microbiology Center of China Microorganism Collection Management Committee, preservation date: 2010-04-22, classification name: hybridoma cell line.

[0025] Below in conjunction with specific embodiment, the present invention is described in further detail:

[0026] The technical scheme of the present invention is as follows:

[0027] 1. Establishment of cell lines

[0028] 1. Antigen Preparation

[0029] a) Expression of African swine fever P54 protein

[0030] According to the African swine fever (ASFV) P54 gene sequence in GenBank, a pair of primers was designed and synthesized. The P54 gene fragment was amplified from ASFV DNA by PCR method, and cloned into the expression vector pET28b to construct the recombinant plasmid pET-P54 , and transformed into the expression host strain BL21(DE3) after sequencing verification, and the expression wa...

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Abstract

The invention discloses a competitive ELISA (Enzyme-Linked Immuno Sorbent Assay)kit for detecting an antibody of an African swine fever virus and application thereof, belonging to the technical field of organisms. The kit is used for detecting an antibody of the African swine fever virus in pig serum by adopting prokaryotically expressed recombinant P54 protein as an envelope antigen according toa competitive ELISA principle. The envelope antigen in a 96 pore plate in the kit is prokaryotically expressed recombinant P54 protein and has favorable antigenicity. The enzyme-linked immuno kit provided by the invention comprises the P54 protein enveloped 96 pore plate, positive control, negative control, a horseradish peroxidase marked monoclonal antibody, a concentrated cleaning solution, serum diluent, a TMB substrate and a stopping solution. The kit can be used for screening samples in bulk, main reagents in the kit are provided in a working solution way, and the use is convenient.

Description

technical field [0001] The invention is a detection kit for detecting African swine fever virus (ASFV) antibody, which belongs to the field of biotechnology. Specifically, the present invention relates to an ELISA kit for detecting antibodies in animal quarantine and use thereof. Background technique [0002] African swine fever (ASF) is an acute, febrile and 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 mortality rate, which can be as high as 100%. The clinical symptoms and pathological changes are similar to acute swine fever. It is easy to be misdiagnosed at the time of diagnosis, and manifests as high fever, skin congestion and cyanosis, miscarriage, edema and organ hemorrhage. (William, Hess Adv. African swine fever: a reassessment [J]. Vet Sci Comp Med, 1981, 25: 39-69) The World Organization for Animals (OIE) lists it as a Class A disease, and my country defines it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
Inventor 董志珍王涛肖妍赵祥平侯艳梅张瑞
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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