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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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 economic losses to the animal breeding industry in Tibet.
These methods are not suitable for on-site testing, rapid testing and clinical testing in grassroots veterinary departments

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

Examples

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Effect test

Embodiment 1

[0027] A monoclonal antibody to the N protein of Peste des petits ruminants virus, said monoclonal antibody to the N protein of Peste des petits ruminants virus is produced by the monoclonal antibody hybridoma cell line 117-6H6 of Peste des pets ruminants virus, and the monoclonal antibody of Peste des pets ruminants virus is hybridized The preservation number of tumor cell line 117-6H6 is 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 the N protein of Peste des petits ruminants virus, comprising the following steps: immunizing mice with purified Peste des petits ruminants virus, taking splenocytes from the immunized mice and SP2 / 0 myeloma cells for cell fusion, and using purified Peste des petits ruminants virus antigen, genetically engineered Peste des pets ruminants virus N protein, and BHK cells to establish indirect ELISA to detect cell culture supernata...

Embodiment 2

[0050] Example 2 Specificity, sensitivity and coincidence rate test of a fluorescent immunochromatographic detection test strip for Peste des petits ruminants virus of the present invention

[0051] (1) Specificity of test strips: Use test strips to detect standard positive samples of Peste des petits ruminant virus (PPRV), other related viruses including bluetongue virus (BTV), type O foot-and-mouth disease virus (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 result shows that the test strip is positive when detecting the PPRV positive sample, and is negative when other virus positive and negative control samples are tested. It shows that the specificity of the test strip is good.

[0052] (2) Sensitivity of test strips: detect serial dilutions (10 1 ~10 7 ) of the PPRV positive samples, while using the RT-PCR metho...

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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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