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A kind of anti-h7 subtype avian influenza virus monoclonal antibody and its application

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

Active Publication Date: 2017-09-29
JIANGSU ACAD OF AGRI SCI +1
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  • Summary
  • 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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  • A kind of anti-h7 subtype avian influenza virus monoclonal antibody and its application
  • A kind of anti-h7 subtype avian influenza virus monoclonal antibody and its application
  • A kind of anti-h7 subtype avian influenza virus monoclonal antibody and its application

Examples

Experimental program
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Effect test

Embodiment Construction

[0021] (1) Establishment of monoclonal antibody hybridoma cell lines

[0022] 1 The preparation of H7 antigen is used in the laboratory of Jiangsu Provincial Center for Disease Control and Prevention, collected from Wuxi Chaoyang Poultry Wholesale Market, RT-PCR tested positive for H7-AIV pigeon throat swabs, and inoculated into 10-day-old SPF chicken embryos after aseptic treatment , Observe the death of chicken embryos. 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.

[0023] Animal immunization A...

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Abstract

The invention relates to an anti-H7 subtype avian influenza virus monoclonal antibody and application thereof, belonging to the field of biotechnology. The present invention screens a hybridoma cell line 5D2, and the hemagglutination inhibition (HI) titer of the mouse ascites antibody prepared by it against H7-AIV is 11 log2, and the ELISA titer is 2×105. The anti-H7-AIV monoclonal antibody secreted by the hybridoma cell line of the present invention has high HI and ELISA titers, and the colloidal gold immunochromatography detection test strip assembled with it has strong specificity, except that H7-AIV shows positive results. , other viruses in poultry are all negative, and can be used for on-site diagnosis of poultry or mammals infected with H7-AIV. The results are available within 10 minutes. The results are intuitive and can be easily judged by naked eyes. The diagnostic kit assembled from the antibody secreted by the hybridoma cell line of the present invention has good stability without lowering sensitivity after being stored for one year.

Description

Technical field [0001] The invention relates to an anti-H7 subtype avian influenza virus monoclonal antibody and its application, belonging 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 both avian and mammalian H7-AI strains, and can be used for rapid on-site diagnosis of H7-AI diseased poultry and mammals. Background technique [0002] Human infection with H7N9 avian influenza is an acute respiratory infectious disease caused by the H7N9 avian influenza virus. Severe pneumonia cases 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 the H7 subtype avian influenza virus (H7-AIV). Human infection with H7N9 avian influenza is an acute respiratory infectious disease caused by the H7N9 avia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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 ACAD OF AGRI SCI
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