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Indirect ELISA reagent kit for detecting antibodies against rabbit hemorrhagic disease virus

A technology of rabbit haemorrhagic disease virus and kit, applied in the direction of measuring device, color/spectral characteristic measurement, instrument, etc., can solve the problems of loose poison, non-specificity, poor repeatability, etc., and achieve good reactogenicity, Guaranteed highly sensitive results

Inactive Publication Date: 2009-09-16
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

The hemagglutination inhibition method (HI) often has poor reproducibility due to the influence of various factors, and the ELISA using virus as the coated antigen is often prone to non-specificity due to factors such as poor virus purification, and there is a potential for using RHDV as the coated antigen. Dangers of poisoning

Method used

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  • Indirect ELISA reagent kit for detecting antibodies against rabbit hemorrhagic disease virus
  • Indirect ELISA reagent kit for detecting antibodies against rabbit hemorrhagic disease virus
  • Indirect ELISA reagent kit for detecting antibodies against rabbit hemorrhagic disease virus

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Embodiment Construction

[0047] 1 Preparation and identification of coated antigen

[0048] 1) Use recombinant virus rAcV-Bac-VP60 (known and public, see "Expression of Rabbit Hemorrhagic Disease Virus Capsid Protein in Insect Cells and Immunoprotective Effect on Rabbits". Journal of Animal Husbandry and Veterinary Medicine, 2008, 39 (10), 1382~1387.), infect Sf9 insect cells (purchased from Invitrogen), and collect cells and culture medium 7 days after infection;

[0049] 2) Freeze and thaw the above collection at -20°C three times, centrifuge at 10,000rpm / min for 10min, collect the supernatant, filter the supernatant with a 0.45μm filter membrane, and detect the supernatant with a hemagglutination test (HA) The hemagglutination titer of the recombinant VP60 protein in the medium was 13log2, and the measured protein concentration was 591mg / L. After aliquoting, it was stored at -20°C for later use;

[0050] 3) The recombinant VP60 protein was identified by protein electrophoresis and Western-blotting...

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Abstract

The invention relates to an ELISA reagent kit for detecting antibodies against rabbit hemorrhagic disease virus (RHDV), and belongs to the field of biotechnology. The reagent kit adopts recombined VP60 capsid protein expressed by a baculovirus expression system as envelope antigen, detects the antibodies against rabbit hemorrhagic disease virus in serum of a rabbit according to an indirect ELISA principle, and is used for rabbit hemorrhagic disease vaccine serological detection in a rabbit group. The core substance envelope antigen of the reagent kit is the recombined VP60 capsid protein expressed by the baculovirus expression system, which has good antigenicity like natural RHDV but no potential poison spreading danger like the RHDV. The reagent kit used to detect serum samples has the advantages of low background reaction and obvious contrast of OD values of negative and positive serum development. The reagent kit has low production cost, strong specificity and high sensitivity, can detect samples in a large quantity, and has important application value in aspects such as rabbit hemorrhagic disease immunity monitoring and the like.

Description

A technical field [0001] The invention is a detection kit for detecting rabbit hemorrhagic disease virus (RHDV) antibody, which belongs to the technical field of veterinary biology. It is used for the detection of rabbit haemorrhagic disease virus antibody, so as to monitor the immune status of rabbit haemorrhagic disease vaccine in rabbit groups, and provide technical basis for the immunization of rabbit haemorrhagic disease vaccine. Two background technology [0002] Rabbit hemorrhagic disease (RHD), also known as rabbit plague, is a rabbit infectious disease characterized by acute, highly contagious, and large-scale death caused by rabbit hemorrhagic disease virus (RHDV). Infected rabbits usually die within 48 to 72 hours. The main features are highly infectious, respiratory system hemorrhage, liver necrosis, visceral edema, congestion and hemorrhage and other changes, which have caused huge economic losses to the rabbit industry all over the world. The International Vet...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543G01N21/31
Inventor 王芳胡波范志宇苏国清蔡少平徐为中张则斌薛家兵何孔旺
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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