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An indirect immunofluorescence detection kit for mink enteritis parvovirus

An enteritis parvovirus and immunofluorescence detection technology, applied in the field of immunology, can solve the problems of short storage period, prone to false positives, unsuitable for distinguishing infection, etc., and achieve the effect of improving specificity

Active Publication Date: 2017-07-28
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the traditional method is practical, it also has disadvantages. The diagnostic method of hemagglutination test has the advantages of simplicity and rapidity.
However, due to the presence of non-hemagglutination strain MEV-S in MEV, there is a possibility of missed detection by this method.
In addition, the pig or monkey red blood cells used in the diagnosis must be fresh red blood cells, not only the sensitivity of HA is greatly affected by the blood donor, but also its short storage period, which limits the popularization and application of this diagnostic method
Virus isolation and identification is a commonly used identification method in the laboratory, but this method takes a long time. It is often encountered that after MEV is inoculated with sensitive cells CRFK or F81, there may not be obvious wire-like lesions visible to the naked eye, and the lesions should not be compared with Differentiation of infection by other viruses of the genus Parvovirus such as mink Aleutian virus
The PCR method is used to detect the nucleic acid of the virus, which has high sensitivity and specificity, but is prone to false positives

Method used

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  • An indirect immunofluorescence detection kit for mink enteritis parvovirus
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  • An indirect immunofluorescence detection kit for mink enteritis parvovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of Mink Enteritis Parvovirus Monoclonal Antibody

[0043] 1. Materials and methods

[0044] 1.1 Cells, virus species, experimental animals and reagents

[0045] Mink enteritis parvovirus SMPV-11 strain, canine distemper virus (CDV) and mink Aleutian virus (ADV) are all preserved by the Economic Animal Disease Research Office of the Institute of Special Products, Chinese Academy of Agricultural Sciences. Mouse myeloma cells (SP2 / 0) were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; BALB / c pure line mice were purchased from the Experimental Animal Center of Changchun Institute of Biological Products.

[0046] Freund's complete adjuvant, Freund's incomplete adjuvant, HAT, and HT selection medium are all products of SIGMA; DMEM medium is a product of GIBCO; horseradish enzyme-labeled goat anti-mouse IgG, FITC-labeled goat anti-mouse IgG was purchased from Beijing Zhongshan Jinqiao...

Embodiment 2

[0080] Example 2 Establishment of Indirect Immunofluorescence Detection Method for Monoclonal Antibody ZL1

[0081] 1 Materials and methods

[0082] 1.1 Test materials and reagents

[0083] The commercially available monoclonal antibody against canine parvovirus was purchased from Beijing Shiji Yuanheng Animal Epidemic Prevention Technology Co., Ltd.; the goat anti-mouse FITC-IgG was purchased from KPL, USA.

[0084] TBST buffer: TBS buffer containing 1‰ Tween20;

[0085] The preparation method of TBS buffer solution: NaCl 8g, KCl 0.2g, Trisbase 3g, pure water 800mL. Adjust the pH to 7.4 with HCl, and add water to make up to 1000 mL.

[0086] 1.2 Determination of biological activity of monoclonal antibody ZL1

[0087] The titer of the monoclonal antibody was detected by the HI experiment according to the conventional method. The ascites obtained from mice injected with SP2 / 0 cells was used as a negative control, the standard MEV serum stored in the room was used as a posit...

Embodiment 3

[0103] The assembly of embodiment 3 kits

[0104] The test kit described in the present invention comprises:

[0105] Slides and cell culture plates: used for smears of samples to be tested or cell-coated cell culture plates inoculated with sterile-filtered suspensions of samples to be tested;

[0106] Anti-mink enteritis parvovirus monoclonal antibody: the monoclonal antibody is produced by the hybridoma cell line with the preservation number CGMCC NO.10881;

[0107] Diluent: the diluent includes anti-mink enteritis parvovirus monoclonal antibody dilution and goat anti-mouse FITC-IgG dilution, the anti-mink enteritis parvovirus monoclonal antibody dilution is TBS buffer containing 3% inactivated goat serum, Goat anti-mouse FITC-IgG dilution is TBS buffer containing 3% inactivated goat serum and 1‰ Evans blue;

[0108] Washing solution: TBST buffer containing 0.3M glycine;

[0109] Positive control: MEVB strain inoculated on a cell culture plate to grow as a monolayer of F8...

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Abstract

The invention discloses an indirect immunofluorescence detection kit for mink enteritis parvovirus. The kit includes the anti-mink enteritis parvovirus monoclonal antibody secreted by the hybridoma cell line with the preservation number CGMCC NO.10881. The invention also discloses an indirect immunofluorescence detection method for rapid detection of mink enteritis parvovirus established by using the monoclonal antibody. The method improves the specificity of the diagnosis of the disease and can realize the specific diagnosis of the disease. Sites of viral infection of cells are directly observed at the cellular level. The establishment of MEV indirect immunofluorescence detection method provides a new method for the diagnosis of MEV.

Description

technical field [0001] The invention relates to a kit, in particular to an indirect immunofluorescence detection kit for mink enteritis parvovirus, which belongs to the field of immunology. Background technique [0002] Mink parvovirus enteritis is an acute and severe infectious disease caused by Minkenteritis virus (MEV) of the Parvoviridae subfamily Mink enteritis virus (MEV) infecting mink with high morbidity and mortality. One of the great diseases. The disease first appeared in Fort William, Canada in 1947, and then occurred in various breeding countries. The disease was first discovered in my country in 1974. In recent years, with the improvement of people's living standards, the mink breeding industry has flourished, and the demand for mink species is increasing day by day. A large number of mink are imported from abroad every year, and domestic mink farms frequently adjust mink species. In addition, the breeding environment is not properly managed. There are inact...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569C12N5/20C07K16/08C12R1/91
CPCC07K16/081G01N33/56983
Inventor 张蕾张海玲胡博白雪刘昊赵建军薛向红吕爽鲁荣光史宁徐淑娟朱言柱闫喜军
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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