Peste des petitis ruminants virus N protein monoclonal antibody, test strip comprising same and preparation method thereof

A technology of Peste des petits ruminants and monoclonal antibodies, applied in the direction of antiviral immunoglobulin, microbe-based methods, biochemical equipment and methods, etc., can solve the problems of economic losses in the animal breeding industry in Tibet, and achieve easy storage and transportation, Detects effects quickly, with good specificity and sensitivity

Inactive Publication Date: 2015-11-04
杨俊兴
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  • Application Information

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Problems solved by technology

In 2007, the disease first broke out in the Ngari area of ​​Tibet, my country. In the following years, the disease still occurred in some areas of Tibet, causing huge econom

Method used

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  • Peste des petitis ruminants virus N protein monoclonal antibody, test strip comprising same and preparation method thereof
  • Peste des petitis ruminants virus N protein monoclonal antibody, test strip comprising same and preparation method thereof

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[0026] Example 1

[0027] A monoclonal antibody against the PPR virus N protein, said PPR virus N protein monoclonal antibody is produced by the PPR monoclonal antibody hybridoma cell strain 117-6H6, and the PPR monoclonal antibody is hybridized The tumor cell line 117-6H6 was deposited as CCTCC NO.C2014233. It was deposited in the China Center for Type Culture Collection on December 3, 2014.

[0028] A method for preparing a monoclonal antibody against PPR virus N protein, comprising the following steps: immunizing mice with purified PPR virus, taking spleen cells of the immunized mice and performing cell fusion with SP2 / 0 myeloma cells, and using purified Indirect ELISA was established to detect the cell culture supernatant by establishing an indirect ELISA to detect the PPR virus antigen, genetically engineered expression of the PPR virus N protein and BHK cells. The three ELISA methods were named: PPRV-ELISA, PPRV N-ELISA and BHK-ELISA, Screen the cells that are positive...

Example Embodiment

[0050] Example 2 Specificity, sensitivity and coincidence rate test of a kind of PPR virus fluorescence immunochromatographic detection test strip of the present invention

[0051] (1) The specificity of the test strip: use the test strip to detect the standard positive samples of Peste des petits ruminants virus (PPRV), other related viruses including bluetongue virus (BTV), foot-and-mouth disease virus type O (FMDV O), deer Epidemic hemorrhagic disease virus (EHDV), Akabane disease virus (AKV), vesicular stomatitis virus (VSV), bovine viral diarrhea virus (BVDV) positive samples and negative control samples. The results showed that the test strip was positive when detecting PPRV positive samples, and it was negative when compared with other virus positive and negative control samples. It shows that the specificity of the test strip is good.

[0052] (2) Sensitivity of test strips: use test strips to detect serial dilutions (10 1 ~10 7 ) PPRV positive samples, and RT-PCR m...

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Abstract

The invention belongs to the technical field of animal virus detection and particularly relates to a peste des petitis ruminants virus N protein monoclonal antibody, a test strip comprising the same and a preparation method thereof. The test strip comprises backing, a sample pad, a fluorescent microsphere-marked peste des petitis ruminants virus monoclonal antibody composite pad, a nitrocellulose membrane with spotted detection line and control line and a water absorption pad, wherein the detection line is coated with a goat anti-peste des petitis ruminants virus polyclonal antibody, the control line is coated with staphylococcus A protein, the peste des petitis ruminants virus monoclonal antibody is the peste des petitis ruminants virus N protein monoclonal antibody which is generated by a peste des petitis ruminants virus monoclonal antibody hybridoma cell strain 117-6H6, and the preservation number of the peste des petitis ruminants virus monoclonal antibody hybridoma cell strain 117-6H6 is CCTCC NO. C2014233. The established des petitis ruminants virus fluorescent immunochromatographic test strip detection test strip can be used for rapid detection and field detection of PPRV, so that the situation that PPRV rapid detection and field detection methods and detection tools are lacking at home is solved.

Description

technical field [0001] The invention belongs to the technical field of animal virus detection, and in particular relates to a monoclonal antibody of N protein of Peste des Petits Ruminants virus, a test strip containing the antibody and a preparation method thereof. Background technique [0002] Peste des petits ruminants (PPR), also known as pseudorinderpest in small ruminants, pneumonitis, stomatitis pneumonia complex, is an acute, severe infectious disease. The World Organization for Animal Health (OIE) lists PPR as an animal disease that must be reported, and the Ministry of Agriculture of my country lists PPR as a first-class animal disease. PPR has seriously affected the healthy development of the ruminant breeding industry and the international trade of related animals and animal products, causing huge economic losses to the affected areas. PPR has the characteristics of high morbidity and mortality. Susceptible animals are mainly goats and sheep, and wild small rum...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N5/20G01N33/577G01N33/569C12R1/91
Inventor 杨俊兴孙洁连慧香花群义吕建强刘建利张彩虹廖立珊曹琛福陈兵阮周曦唐金明
Owner 杨俊兴
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