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Cyprinid herpesvirus II type sandwich ELISA detection kit and detection method

A carp herpes virus and detection kit technology, which is applied in the field of carp herpes virus type II sandwich ELISA detection kits, can solve the problem of inability to confirm whether there are infectious virus particles in fish and the like

Inactive Publication Date: 2017-05-31
LIANYUN PORT IMMIGRATION INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, PCR or quantitative PCR method is commonly used at home and abroad to detect carp herpes virus type Ⅱ. There is latent infection of herpes virus. Fish with healthy appearance may carry the virus. PCR method can detect the virus nucleic acid, but it cannot confirm whether there are infectious virus particles in the fish. , so it is necessary to establish an immunological method for the detection of viral proteins

Method used

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  • Cyprinid herpesvirus II type sandwich ELISA detection kit and detection method
  • Cyprinid herpesvirus II type sandwich ELISA detection kit and detection method
  • Cyprinid herpesvirus II type sandwich ELISA detection kit and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0049] Embodiment 1, a kind of carp herpes virus type Ⅱ sandwich ELISA detection kit, the main components of this kit are:

[0050] Cyprin herpesvirus type II ORF90 purified monoclonal antibody solution, 1 tube 100 μL, is a solution made of carp herpesvirus type II ORF90 purified monoclonal antibody dissolved in water, the concentration is 10-20 mg / mL;

[0051] Enzyme-labeled ORF90 polyclonal antibody solution, 1 bottle of 100 μL, is a solution made of enzyme-labeled ORF90 polyantibody dissolved in water, the concentration is 10-20mg / mL;

[0052] Diluent, 2 bottles, 50 mL each, the solution is bovine serum albumin phosphate buffer with a mass volume concentration of 10 g / L;

[0053] TMB substrate solution, 1 tube, 250 μL; it is the DMSO solution of TMB, its mass volume concentration

[0054] 6 mg / mL;

[0055] Stop solution, 1 bottle of 2 M sulfuric acid solution, 20 mL;

[0056] Carp herpesvirus type 2 positive control 1 tube, 500 μL, which is goldfish positive tissue homog...

Embodiment 2

[0060]Example 2, in a kind of carp herpesvirus type II sandwich ELISA detection kit described in Example 1: the weight-to-volume ratio of the phosphate buffer consists of:

[0061] Sodium chloride 8.0 g

[0062] Potassium dihydrogen phosphate 0.2 g

[0063] Disodium hydrogen phosphate dodecahydrate 2.9 g

[0064] Potassium chloride 0.2g

[0065] Tween-80 0.5 mL

[0066] Double distilled water 1000 mL.

Embodiment 3

[0067] Embodiment 3, in a kind of carp herpesvirus type II sandwich ELISA detection kit described in embodiment 1 or 2: the kit also includes:

[0068] Phosphate buffer saline PBST, 1 bottle of 50 mL, its mass volume ratio composition is:

[0069] Sodium chloride 8.0 g

[0070] Potassium dihydrogen phosphate 0.2 g

[0071] Disodium hydrogen phosphate dodecahydrate 2.9 g

[0072] Potassium chloride 0.2 g

[0073] Tween-80 0.5 mL

[0074] Double distilled water 1000 mL.

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Abstract

The invention relates to a cyprinid herpesvirus II type sandwich ELISA detection kit. The kit mainly comprises a cyprinid herpesvirus II type ORF90 purified monoclonal antibody solution, an enzyme-labeled ORF90 polyclonal body solution, a diluent, a TMB substrate solution, a stop solution, cyprinid herpesvirus II type positive control, negative control, coating liquid and a 96-pore enzyme-labeled plate. The invention also relates to a detection method. The kit and the method, provided by the invention, can be applied to detection of latent infected fish virus without clinical symptom and can further be applied to research on the change rule of the cyprinid herpesvirus II type in the fish body, so that basis is provided for prevention and control of the virus. The method provided by the invention does not have specific reaction with other common viruses of crucian and goldfish, and can be applied to diagnosis of epidemic disease in domestic farms as well as exit and entry fish quarantine.

Description

technical field [0001] The invention relates to a detection kit, in particular to a carp herpesvirus type II sandwich ELISA detection kit, and also relates to a detection method using the detection kit. Background technique [0002] In recent years, Cyprinid herpesvirus Ⅱ (English name: Cyprinid herpesvirus Ⅱ, abbreviated as CyHV-2) has brought great harm to the crucian carp and goldfish aquaculture industry in my country. The morbidity and mortality are extremely high when the disease breaks out. According to incomplete statistics In 2006, the economic loss caused by the disease was as high as 718 million yuan. [0003] In the prior art, the most intuitive method for its diagnosis is to conduct histological examination on diseased fish tissues and then observe herpes virus particles by electron microscope for confirmation. At present, PCR or quantitative PCR method is commonly used at home and abroad to detect carp herpes virus type Ⅱ. There is latent infection of herpes vi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/569G01N33/577
CPCG01N33/535G01N33/543G01N33/56994G01N33/577
Inventor 徐晔段宏安周毅丁超曹洁王凤芝程积进
Owner LIANYUN PORT IMMIGRATION INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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