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Anti-H7 subtype avian influenza virus monoclonal antibody and application thereof

A technology of monoclonal antibody and avian influenza virus, applied in the direction of antiviral immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problem of time-consuming nucleic acid detection and achieve intuitive results

Active Publication Date: 2015-01-28
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

"Diagnosis and Treatment Program for Human Infection with H7N9 Avian Influenza" (2014 Edition) clearly stated that "nucleic acid testing should be preferred in early identification of cases", but nucleic acid testing takes a long time

Method used

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  • Anti-H7 subtype avian influenza virus monoclonal antibody and application thereof
  • Anti-H7 subtype avian influenza virus monoclonal antibody and application thereof
  • Anti-H7 subtype avian influenza virus monoclonal antibody and application thereof

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Embodiment Construction

[0024] (1) Establishment of monoclonal antibody hybridoma cell lines

[0025] 1 Preparation of H7 antigen Use the laboratory of Jiangsu Provincial Center for Disease Control and Prevention to collect pigeon swabs from Chaoyang Poultry Wholesale Market in Wuxi, and RT-PCR test to be H7-AIV positive pigeon throat swabs. After aseptic treatment, they are inoculated into 10-day-old SPF chicken embryos and observe the death of chicken embryos. Happening. Excluding dead embryos within 24 hours, chicken embryos that have not died within 96 hours were frozen at -20°C, and frozen for 2 hours, and then allantoic fluid was collected. Centrifuge the allantoic fluid at 8,000 rpm at 4°C for 30 min. Take the supernatant and place it in an ultracentrifuge tube. Centrifuge at 36,000 rpm at 4°C for 2 h. Discard the supernatant. Resuspend the pellet in physiological saline and perform spectroscopy. Determine the virus concentration with a photometer, and save it for later use after inactivation....

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Abstract

The invention relates to an anti-H7 subtype avian influenza virus monoclonal antibody and application thereof, belonging to the technical field of biology. A hybridoma cell strain 5D2 is screened; the hemagglutination inhibition (HI) titer of the rat ascites antibody prepared from the hybridoma cell strain 5D2 for H7-AIV is 11log2, and the ELISA titer is 2*10<5>. The anti-H7 monoclonal antibody secreted by the hybridoma cell strain has high HI and ELISA titer; the colloidal gold immunochromatography detection test strip assembled by the anti-H7 monoclonal antibody has high specificity, except that the result for H7-AIV is positive, the results for the other avian viruses are negative; the anti-H7 monoclonal antibody can be used for on-site diagnosis on fowl, mammals or the like infected by H7-AIV; and the result can come out within 10 minutes, is visual, and can be easily judged with naked eyes. The sensitivity of the diagnosis kit assembled by the hybridoma cell strain secretory antibody is not lowered after the diagnosis kit is stored for one year, so the diagnosis kit has favorable stability.

Description

technical field [0001] The invention relates to an anti-H7 subtype avian influenza virus monoclonal antibody and application thereof, belongs to the field of biotechnology, and relates to antibody engineering technology. Specifically, it is an anti-H7-AIV high-titer monoclonal antibody, which has high titer for H7-AI strains of both poultry and mammalian origin, and can be used for on-site rapid diagnosis of H7-AI-affected poultry and mammals. Background technique [0002] Human infection with H7N9 avian influenza is an acute respiratory infectious disease caused by H7N9 avian influenza virus, among which cases of severe pneumonia can often be combined with acute respiratory distress syndrome, septic shock, and even multiple organ failure. [0003] H7N9 subtype avian influenza (H7-AI) is a respiratory infectious disease of poultry and mammals caused by H7 subtype avian influenza virus (H7-AIV). Human infection with H7N9 avian influenza is an acute respiratory infectious di...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/10G01N33/577C12R1/91
Inventor 李银刘晓燕赵冬敏黄欣梅杨婧刘宇卓韩凯凯刘岳龙戴建君祁贤刘青涛谢星星
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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