Cell line resistant to A subgroup avian leukosis virus as well as construction method and application thereof

A technology of avian leukemia virus and construction method, which is applied in the field of cell line and construction of avian leukemia virus resistant to subgroup A, and can solve the problems of inability to distinguish endogenous and exogenous, rarity limiting efficacy and the like

Active Publication Date: 2015-04-29
SOUTH CHINA AGRI UNIV
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Problems solved by technology

[0003] At present, domestic and foreign detection and identification methods for the pathogen of avian leukemia mainly include cell culture, ELISA, indirect immunofluorescence test, PCR, etc. Among them, ELISA is the most widely used clinically, and its advantage is that the method is relatively simple to operate and commercialized. Detection kits are available, but the disadvantage is that endogenous and exogenous ALV cannot be distinguished; indirect immunofluore

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  • Cell line resistant to A subgroup avian leukosis virus as well as construction method and application thereof
  • Cell line resistant to A subgroup avian leukosis virus as well as construction method and application thereof
  • Cell line resistant to A subgroup avian leukosis virus as well as construction method and application thereof

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[0028] The present invention will be further described in detail below in conjunction with the examples and drawings, but the implementation of the present invention is not limited to this.

[0029] The present invention adopts the following technical solutions:

[0030] DF-1 cell line: from the Key Laboratory of Veterinary Vaccine Creation, Ministry of Agriculture.

[0031] Strains: RAV-1 and GD13 are ALV-A, CHN06 is ALV-J, provided by the Key Laboratory of Veterinary Vaccine Creation of the Ministry of Agriculture.

[0032] Enzymes: Restriction enzymes BamHI, NotI and T4 ligase were purchased from NEB Company; LA Taq was purchased from TaKaRa Company.

[0033] Plasmid: eukaryotic expression plasmid pcDNA3.1 / Zeo(+) was purchased from Invitrogen; RCAS(A) plasmid, gifted by ADOL laboratory in the United States.

[0034] Antibody: ALV-A specific monoclonal antibody, gift from ADOL laboratory in the United States. FITC-labeled goat anti-mouse IgG antibody and HRP-labeled goat anti-mouse Ig...

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Abstract

The invention discloses a cell line resistant to an A subgroup avian leukosis virus as well as a construction method and application thereof, and belongs to the technical field of agricultural biology. The cell line resistant to the A subgroup avian leukosis virus is named after an anti ALV-A chicken embryo fibroblast cell line DF-1/A, and is conserved in Wuhan University, China of China Center for Type Culture Collection (CCTCC) with the preservation number of CCTCC NO: C2014180 from October 15th, 2014; the cell line resistant to the A subgroup avian leukosis virus is applied to the diagnosis of the A subgroup avian leukosis virus. The cell line resistant to the A subgroup avian leukosis virus disclosed by the invention can achieve stable passage and be stored for a long term, is helpful for accurately identifying subgroup classification of ALV by using the characteristic of ALV superinfection, and is applied to the diagnosis of the A subgroup avian leukosis virus.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a cell line against subgroup A avian leukosis virus and its construction method and application. Background technique [0002] At present, avian leukemia is seriously harmful to my country's poultry industry, and various pathological phenotypes such as lymphoma, myeloid cell tumor type and hemangioma type have appeared. and other neoplastic diseases. The disease is mainly transmitted vertically. Effective measures to control the disease are to reduce the infection rate of breeder flocks and establish leukemia-free breeder flocks. Eliminate virus-positive chickens to purify and eliminate ALV in chicken flocks. Among the exogenous ALVs, the J subgroup, A subgroup and B subgroup avian leukosis virus have obvious pathogenicity to chickens, and are also very harmful to poultry production. [0003] At present, domestic and foreign methods for the detection and id...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12N15/85C12Q1/70C12Q1/68C12R1/93
Inventor 曹伟胜冯敏代曼曼廖明
Owner SOUTH CHINA AGRI UNIV
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