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Hemagglutination inhibition test antigen for duck tembusu virus disease and preparation method thereof

A technology of duck Tembusu virus and hemagglutination inhibition test, which is applied in the field of bioengineering and immunology, can solve the problems of unified regulations or formulation of the disease name, and no country yet, and achieve the effect of antigen stabilization

Active Publication Date: 2016-07-27
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2011, the first waterfowl disease prevention and control seminar of the Chinese Association of Animal Husbandry and Veterinary Medicine unified the name of the disease as "Duck Tembusu Virus Disease", but so far, there is no unified regulation or formulation on the name of the disease at the national level

Method used

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  • Hemagglutination inhibition test antigen for duck tembusu virus disease and preparation method thereof
  • Hemagglutination inhibition test antigen for duck tembusu virus disease and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Raw materials for the preparation of antigens:

[0026] 1) Sensitive (choose 2-4 days old closed group or inbred strain) suckling mice, via intracerebral route, according to 25μl / mouse (about 100ELD 50 ) dose inoculated with duck Tembusu virus, observed to 144 hours, the suckling mice appeared clinical symptoms such as mental depression and tremor, and the suckling mice morbidity and mortality should be greater than 90%.

[0027] 2) Duck Tembusu virus, a member of the Flaviviridae family, contains a specific gene sequence, and its GenBank accession number is JF523187. The results of gene sequence analysis of the isolated strain show that the strain has the highest homology with Tembusu virus. Virus particles are 30-50nm in size, spherical, with capsules and fibrils, and can agglutinate 0.25-0.6% goose and pigeon red blood cells. The virus can be detected in closed population or inbred mouse brain, chicken embryo, duck embryo, duck embryo fibroblast, C6 / 36 (Aedes albopi...

Embodiment 2

[0045] Raw materials for the preparation of antigens:

[0046] 1) C6 / 36 cells (100TCID 50 / 0.1ml dose to inoculate the cells growing into a monolayer, and the typical CPE lesions appeared in the cells after 120 hours of observation).

[0047] 2) Duck Tembusu virus, a member of the Flaviviridae family, contains a specific gene sequence, and its GenBank accession number is JF523187. The results of gene sequence analysis of the isolated strain show that the strain has the highest homology with Tembusu virus. Virus particles are 30-50nm in size, spherical, with capsules and fibrils, and can agglutinate 0.25-0.5% goose and pigeon red blood cells. The virus can be detected in closed population or inbred mouse brain, chicken embryo, duck embryo, duck embryo fibroblast, C6 / 36 (Aedes albopictus cells), BHK21 (suckling hamster kidney cells) and Vero (African green monkey kidney cells). ) grow on the cells.

[0048] The virus mainly infects ducks, geese and chickens, etc. Ducks, geese...

Embodiment 3

[0065] On the basis of Example 1, there is further optimization, and the materials and reagents used are the same as in Example 1.

[0066] Duck Tembusu virus HB strain (F1 generation) was used as the virus seed, with 25 μl / piece (including 100ELD 50) dose of intracerebral inoculation of 2 to 4-day-old KM breed suckling mice. Take out the mice with obvious clinical symptoms such as depression and tremor in time, and store them below -20°C. The suckling mice were taken out, the brains were disinfected with tincture of iodine, the skin and skull of the brains were removed under aseptic conditions, the brain tissue was taken out, and placed in a sterilized container. Add sterile sucrose solution with a final concentration of 8.5% according to the ratio of W / V=1:4, and homogenize. Under stirring state, add the homogenate dropwise into the cold acetone of 10-30 times the volume of the homogenate. Centrifuge at 500 g for 5 min, discard the supernatant, crush the precipitate with ...

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Abstract

The invention relates to a hemagglutination inhibition test antigen for a duck tembusu virus disease and a preparation method thereof. A duck hemorrhagic ovarian inflammation virus HB strain is preserved at the typical culture preservation centre in China with the preservation number being CCTCC V201122; the strain comprises a specific gene sequene, and the GenBank accession number of the strain is JF523187. The strain is inoculated againt a neonatal rat proliferating virus; the hemagglutination inhibition test antigen for the duck tembusu virus disease is prepared through the technologies of hemagglutination inhibition nonspecific material (lipoprotein) removal, inactivation, addition of a protective agent and the like. The invention further discloses the preparation method of the hemagglutination inhibition test antigen by a duck tembusu virus.

Description

technical field [0001] The invention relates to a hemagglutination inhibition test antigen for duck Tembusu virus disease and a preparation method thereof, and relates to the fields of bioengineering and immunology. Background technique [0002] Duck Tembusu virus disease is a new or sudden infectious disease that occurred in 2010. The main clinical features are a sudden drop in the egg production of laying ducks and a significant increase in the mortality rate of commercial meat ducks. The number of cases of natural infection and artificial infection The main gross lesions were follicle deformation, hyperemia, hemorrhage, and changes in spleen volume (spleen enlargement in the early stage of infection and shrinkage in the later stage) and blackening of color. The main histological lesion is the activation and hyperplasia of lymphocytes and reticular cells. In the early stage of the epidemic, according to the main pathological changes, the disease was tentatively named "duck...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
CPCG01N33/531G01N33/56983
Inventor 刘月焕林健王小蕾杨志远段会娟赵际成刘立新潘洁
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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