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Influenza A virus ELISA nucleoprotein capture antigen diagnose reagent kit and special monoclonal antibody therefor

A monoclonal antibody, influenza virus technology, applied in anti-viral immunoglobulin, biological testing, biochemical equipment and methods, etc., can solve the problems of high cost, time-consuming virus isolation method, unsuitable for large-scale sample detection, etc. To achieve the effect of high potency and specificity, high application value and high sensitivity

Inactive Publication Date: 2009-01-21
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus isolation and RT-PCR are currently the two most commonly used methods, but virus isolation takes a long time and is not easy to use as a rapid diagnosis; RT-PCR, although it takes a short time, is expensive and not easy to use for large-scale investigations
Hemagglutination and hemagglutination inhibition tests need to be based on virus isolation, and it is difficult to implement and promote at the grassroots level, so it is not suitable for large-scale sample testing

Method used

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  • Influenza A virus ELISA nucleoprotein capture antigen diagnose reagent kit and special monoclonal antibody therefor
  • Influenza A virus ELISA nucleoprotein capture antigen diagnose reagent kit and special monoclonal antibody therefor
  • Influenza A virus ELISA nucleoprotein capture antigen diagnose reagent kit and special monoclonal antibody therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Embodiment 1, to the monoclonal antibody secreted by type A influenza virus NP protein hybridoma cell line C4CGMCC No.2575

[0012] 1) Preparation of anti-type A bird influenza virus nucleoprotein monoclonal antibody

[0013] a. Animal immunity

[0014] The H9N2 subtype avian influenza virus inactivated with formaldehyde was injected into Balb / c mice to produce polyclonal antibody serum.

[0015] b. Cell fusion and cloning

[0016] After the test result of the mouse serum was high, the splenocytes were taken and fused with SP2 / 0 myeloma cells at a ratio of 7:1, the cell supernatant was measured by indirect competitive ELISA, and the positive wells were screened. The positive wells were cloned by the limiting dilution method until a hybridoma cell line capable of stably secreting a monoclonal antibody against the nucleoprotein of avian influenza virus was obtained, and the hybridoma cell line of the monoclonal antibody against the nucleoprotein of the avian influenza v...

Embodiment 2

[0021] Embodiment 2, Type A influenza virus nucleoprotein capture ELISA antigen diagnostic kit

[0022] 1. Composition of Influenza A Virus Nucleoprotein Capture ELISA Antigen Diagnostic Kit

[0023] Type A influenza virus nucleoprotein capture ELISA antigen diagnostic kit consists of: 1) a solid carrier coated with anti-type A avian influenza virus nucleoprotein monoclonal antibody; 2) horseradish peroxidase-labeled anti-avian influenza virus nucleus Protein monoclonal antibody; 3) Enzyme substrate storage solution; 4) Negative control and positive control; 5) Diluent of enzyme conjugate and specimen; 6) Washing solution; 7) Enzyme reaction termination solution.

[0024] 1) Preparation of a solid carrier coated with anti-type A bird flu virus nucleoprotein monoclonal antibody

[0025] Dilute the monoclonal antibody secreted by the type A influenza virus NP protein hybridoma cell line C4CGMCC No.2575 with coating buffer to a protein content of 1.5 μg / mL, and bind to a high-ef...

Embodiment 3

[0062] Example 3, using the type A influenza virus nucleoprotein capture ELISA antigen diagnostic kit of embodiment 2 to detect type A influenza virus nucleoprotein antigen

[0063] 1) Determination of specificity

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Abstract

The invention discloses an A-type flu virus nuclein capture ELISA antigen diagnostic reagent kit and a special monoclonal antibody. The special A-type bird flu virus nuclein monoclonal antibody is secreted by an A-type flu virus NP protein hybridoma strain C4 with a collection number of CGMCC No.2575. The reagent kit comprises a solid vector which envelops the monoclonal antibody and the monoclonal antibody which is labeled by enzyme. The special A-type bird flu virus nuclein monoclonal antibody has high titer and specificity. The reagent kit has simple operation, high sensitivity, strong specificity and high application value, and can be used for clinic examination and laboratory examination of A-type flu virus.

Description

technical field [0001] The invention relates to a type A influenza virus nucleoprotein capture ELISA antigen diagnostic kit and a special monoclonal antibody. Background technique [0002] Frequent outbreaks of avian influenza have brought huge economic losses to the aquaculture industry; rapid or early diagnosis of influenza virus and purification of avian influenza in poultry aquaculture are important links in controlling the disease. Influenza A virus subtypes include 16 HA subtypes and 9 NA subtypes. The hemagglutinin antigens of each subtype are quite different, and it is difficult to diagnose subtype antigens for HA. The nucleoprotein (NP) protein has a low degree of variation among various subtypes of influenza A virus and has good conservation, which can be used as a type A specific antigen for diagnosis of influenza A virus. [0003] The methods currently used in the diagnosis of AIV pathogens mainly include virus isolation, hemagglutination and hemagglutination in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10G01N33/577G01N33/68
Inventor 刘金华阎春梅马广鹏蒲娟刘芹防
Owner CHINA AGRI UNIV
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