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CyHV-2 (Cyprinid Herpesvirus 2) monoclonal antibody, hybridoma cell and application of CyHV-2 monoclonal antibody

A monoclonal antibody, hybridoma cell technology, applied in antiviral immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems of inconvenient rapid diagnosis, lag, single detection method, etc., to improve the detection speed and detection. Efficiency, good specificity and potency, highly specific effects

Inactive Publication Date: 2015-05-06
SHENZHEN AUDAQUE DATA TECH
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cell culture isolation techniques are the most accurate diagnostic method, however, studies have found that CyHV-2 is difficult to proliferate in cell lines commonly used for fish virus isolation
In terms of molecular detection, especially the PCR method is currently a more accurate method for detecting CyHV-2 infection because it can detect a very small amount of viral DNA in tissues; Inconvenience
Electron microscope observation is the most direct method, but it also requires special testing equipment, and it is not convenient for on-site rapid diagnostic testing
[0004] The existing detection methods for goldfish herpesvirus hematopoietic necrosis are mainly PCR, histopathology and other methods, and the detection methods are relatively simple; while the immunological research is relatively lacking or lagging behind, there is no immunological detection method for this disease

Method used

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  • CyHV-2 (Cyprinid Herpesvirus 2) monoclonal antibody, hybridoma cell and application of CyHV-2 monoclonal antibody
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  • CyHV-2 (Cyprinid Herpesvirus 2) monoclonal antibody, hybridoma cell and application of CyHV-2 monoclonal antibody

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Experimental program
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Embodiment

[0021] 1. Materials and methods

[0022] 1. Viral Antigens and Reagents

[0023] CyHV-2 antigen: from CyHV-2-infected diseased fish tissue, tissue grinding samples were taken, and purified by sucrose gradient centrifugation. Escherichia coli TG1 was provided by Dai Heping, a researcher at the Institute of Hydrobiology, Chinese Academy of Sciences.

[0024] 2YT medium: Weigh 17g tryptone, 10g yeast extract, 5g NaCl, dissolve in double distilled water, dilute to 1L, and sterilize by high pressure steam. Prepare solid medium, add 15g agar powder, and sterilize with high pressure steam.

[0025] SOBAG medium: Weigh 20g tryptone, 5g yeast extract, 0.5g NaCl, dissolve in double distilled water, and dilute to 1L. With solid medium, add 15g agar powder. Autoclaved. When it is cooled to 40-50°C, add 10 mL of 1M MgCl2 that has been sterilized, so that the Amp is 100 μg / mL and the glucose content is 2%.

[0026] 2. CyHV-2 monoclonal antibody preparation

[0027] 2.1 Immunity

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Abstract

The invention discloses a CyHV-2 (Cyprinid Herpesvirus 2) monoclonal antibody, a hybridoma cell and an application of the CyHV-2 monoclonal antibody. The CyHV-2 monoclonal antibody disclosed by the invention is secreted from a hybridoma cell GFHNV-6G7 with the collection number of CCTCC No. C2013114 or hybridoma cell GFHNV-3B5 with the collection number of CCTCC No. C2013115. The CyHV-2 monoclonal antibody disclosed by the invention has good specificity and potency; by producing rapid immunodetection test paper from the CyHV-2 monoclonal antibody, the speed and efficiency of detection can be increased, and field detection on CyHV-2 can be realized. The CyHV-2 monoclonal antibody disclosed by the invention makes up the blank in immunodetection on the CyHV-2 and lays a foundation for immunological research on the CyHV-2.

Description

technical field [0001] The present application relates to the field of monoclonal antibodies, in particular to the monoclonal antibody for CyHV-2, the hybridoma cells secreting the monoclonal antibody, and the application of the monoclonal antibody. Background technique [0002] Cyprinid herpesvirus 2 (CyHV-2 for short) is the pathogen of herpes virus hematopoietic organ necrosis, which can cause a large number of deaths of farmed goldfish and crucian carp. From 2011 to 2012, the disease broke out in cultured crucian carp in central my country, resulting in extremely high mortality (Wang et al., 2012). In 2011, CyHV-2 was detected in goldfish exported from my country to Malaysia. In the same year, a large number of crucian carp died in an outbreak in Jiangsu Province. The assay was successful and the virus particles were observed in the spleen and kidney. It can be seen that herpes virus hematopoietic organ necrosis has been introduced into my country, and has seriously aff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08G01N33/577G01N33/569C12N5/20
Inventor 何俊强王津津刘荭于力郑晓聪贾鹏兰文升史秀杰
Owner SHENZHEN AUDAQUE DATA TECH
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