Anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and application

A monoclonal antibody and respiratory syndrome technology, applied in antiviral immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems of high cost and long detection time

Active Publication Date: 2011-02-23
CHINA ANIMAL DISEASE CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current detection techniques for porcine reproductive and respiratory syndrome include: virus isolation, immunoperoxidase monolayer cell assay (IPMA), indirect immunofluorescence (IFA), RT-PCR and other methods, but the detection takes a long time and costs expensive

Method used

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  • Anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and application
  • Anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and application
  • Anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the preparation of monoclonal antibody and the mensuration of biological characteristic

[0022] 1. Preparation of monoclonal antibodies

[0023] 1. Virus species and cells

[0024] Porcine reproductive and respiratory syndrome super mutant strain (Nsp21594-1680 mutant strain) NVDC-JXA1CGMCCNo.1964, hereinafter referred to as PRRSV JXA1 strain.

[0025] PRRSV JXA1 strain, Marc-145 cells, and SP2 / 0 cells were preserved by the China Animal Disease Prevention and Control Center.

[0026] 2. Virus propagation and purification

[0027] The PRRSV JXA1 strain was inoculated with Marc-145 cells that had grown into a single layer, adsorbed at 37°C for 1 hour, added maintenance solution, and harvested the virus when more than 75% of the cells showed cytopathic changes (CPE), and the harvested virus liquid was repeatedly frozen and thawed three times to Centrifuge at 6000rpm for 30min, collect the supernatant, add PEG 6000 (final concentration: 8%) to the supernat...

Embodiment 2

[0081] Embodiment 2, the application of monoclonal antibody

[0082] 1. Preparation of colloidal gold rapid detection test strips

[0083] 1) Preparation of colloidal gold conjugate pads: prepare colloidal gold particles according to conventional methods, label the purified monoclonal antibody obtained in Example 1 on the surface of colloidal gold particles, dilute with colloidal gold diluent and spray on the cut glass fiber On membrane, freeze-dried.

[0084] 2) Spray detection membrane: On the same piece of nitrocellulose membrane, spray the purified monoclonal antibody (detection line) and goat anti-mouse IgG (control line) with Bio-Dot film spraying machine, and the distance between the control line and the detection line 3-7mm, put it in an incubator to dry, take it out and keep it sealed for later use.

[0085] 3) Assemble the test strip: the sample absorbent pad (glass fiber membrane), the conjugate pad, the nitrocellulose membrane sprayed with the detection line (T l...

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Abstract

The invention discloses an anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and application. The invention provides an anti-porcine reproductive and respiratory syndrome virus monoclonal antibody hybridoma cell strain 3C3 with collection number of CGMCC No. 4109. The invention also provides the anti-porcine reproductive and respiratory syndrome virus monoclonal antibody generated by the anti-porcine reproductive and respiratory syndrome virus monoclonal antibody hybridoma cell strain 3C3 with collection number of CGMCC No. 4109. An experiment of the invention proves that: a purified PRRSV JXA1 strain is used for immunizing BALB/c mice, one hybridoma cell strain which can stably secrete PRRSV monoclonal antibodies is screened by utilizing the hybridoma technology and the biological characteristic identification is performed on the hybridoma cell strain so as to lay the foundation for further establishing a specific, sensitive and rapid PRRSV detection method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and its application. Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS) is one of the important porcine infectious diseases at present. The disease is characterized by fever, anorexia, infertility, abortion, stillbirth, mummified fetus, weak piglets and piglets in sows. Characterized by respiratory symptoms and high mortality, it is commonly known as "PRRS". The disease first broke out in the United States in 1987, causing serious economic losses and arousing widespread international concern. In 1996, Guo Baoqing and others isolated the virus from diseased pigs in my country and confirmed it as porcine reproductive and respiratory syndrome (PRRSV). At present, the virus has caused endemic pig populations in many provinces and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12R1/91C07K16/10G01N33/577G01N33/569C12N5/20
Inventor 曹振张倩邓小雨遇秀玲王传彬曲萍翟新验田克恭
Owner CHINA ANIMAL DISEASE CONTROL CENT
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