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Rabbit pestivirus antibody rapid detection card and preparation method thereof

A rabbit plague virus and detection card technology, applied in the field of immune detection, can solve the problems of reducing time and cost, unsuitable for long-term storage, expensive red blood cells, etc., and achieve the effect of accurate detection

Active Publication Date: 2016-04-13
SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the detection methods of rabbit plague virus antibody include hemagglutination method, RT-PCR method, and enzyme-linked immunosorbent assay. Among them, RT-PCR method and enzyme-linked immunoassay method require large-scale instruments and professionals, while hemagglutination method is simple to operate, but Human red blood cells are needed, which are expensive and not suitable for long-term storage. This study uses the principle of immune colloidal gold to develop a rapid detection card preparation method for rabbit plague virus antibodies, which reduces the time for rapid, sensitive and accurate detection of rabbit plague virus antibodies. and cost

Method used

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  • Rabbit pestivirus antibody rapid detection card and preparation method thereof
  • Rabbit pestivirus antibody rapid detection card and preparation method thereof
  • Rabbit pestivirus antibody rapid detection card and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1 A kind of rapid detection card preparation method of rabbit plague virus antibody

[0036] (1) Preparation of anti-rabbit plague virus VP60 monoclonal antibody

[0037] ①Immunization program

[0038] Mix insect cell-expressed protein VP60 immunogen (Shandong Binzhou Animal Husbandry and Veterinary Research Institute) and 501 adjuvant (Shandong Lvdu Biotechnology Co., Ltd., product number LD003) at a volume ratio of 1:1, and treat 8-10-week-old Balb / c mice (purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.) were immunized, and the legs were injected intramuscularly, 50 μg / mouse, and then immunized once every two weeks, with the same dose and site as above; Intraperitoneal injection, no adjuvant, double dose;

[0039] ② Cell fusion

[0040] The splenocytes in Balb / c mice were mixed with the myeloma cells in the peritoneal cavity at a ratio of 20:1, centrifuged at 800 rpm for 7 minutes, the supernatant was discarded, 1 mL of 50%...

Embodiment 2

[0057] Embodiment 2 A kind of rapid detection card preparation method of rabbit plague virus antibody

[0058] (1) Preparation of anti-rabbit plague virus VP60 monoclonal antibody

[0059] ①Immunization program

[0060] Mix insect cell-expressed protein VP60 immunogen (gifted by Binzhou Animal Husbandry and Veterinary Research Institute of Shandong Province) and 501 adjuvant (Shandong Lvdu Biotechnology Co., Ltd., product number LD003) at a volume ratio of 1:0.6, for 8-10 weeks Balb / c mice (purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.) were immunized with 50 μg / mouse in the leg muscles, and then immunized once every two weeks. The dose and site of immunization were the same as above; Immunization, intraperitoneal injection, no adjuvant, double dose.

[0061] ② Cell fusion

[0062] The splenocytes in Balb / c mice were mixed with the myeloma cells in the peritoneal cavity according to the ratio of cell number 20:0.7, centrifuged at 800rpm for 8...

Embodiment 3

[0079] Embodiment 3 A kind of rapid detection card preparation method of rabbit plague virus antibody

[0080] (1) Preparation of anti-rabbit plague virus VP60 monoclonal antibody

[0081] ①Immunization program

[0082] Mix insect cell-expressed protein VP60 immunogen (gifted by Binzhou Animal Husbandry and Veterinary Research Institute of Shandong Province) and 501 adjuvant (Shandong Lvdu Biotechnology Co., Ltd., product number LD003) at a volume ratio of 1:1.3 for 8-10 weeks Balb / c mice (purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.) were immunized with 50 μg / mouse in the leg muscles, and then immunized once every two weeks. The dose and site of immunization were the same as above; Immunization, intraperitoneal injection, no adjuvant, double dose;

[0083] ② Cell fusion

[0084] The splenocytes in Balb / c mice were mixed with the myeloma cells in the peritoneal cavity according to the ratio of cell number 20:1.3, centrifuged at 800rpm for 8m...

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Abstract

The invention relates to a rabbit pestivirus antibody rapid detection card and a preparation method thereof. The preparation method comprises the following concrete steps: (1) preparing an anti-rabbit-pestivirus VP60 monoclonal antibody, concretely performing immunization program, cell fusion, hybridomas screening and cloning, and antibody preparation and purification; and (2) preparing the rabbit pestivirus antibody rapid detection card, concretely decocting a colloidal gold solution, marking the anti-rabbit-pestivirus VP60 monoclonal antibody, processing a colloidal-gold pad, processing a sample pad, and determining C and T lines. The rabbit pestivirus antibody rapid detection card is capable of rapidly, sensitively and accurately detecting rabbit pestivirus antibody, and overcomes the problems that domestic rabbit pestivirus antibody detection mainly employs erythrocyte hemagglutination and ELISA detection.

Description

technical field [0001] The invention relates to a rapid detection card for rabbit plague virus antibody and a preparation method thereof, belonging to the technical field of immune detection. Background technique [0002] Rabbit plague is an acute, febrile, septic and devastating infectious disease caused by a virus. It can occur all year round, and all kinds of rabbits are susceptible. Young rabbits and adult rabbits over 3 months old have the highest morbidity and mortality (up to 95%). [0003] At present, the detection methods of rabbit plague virus antibody include hemagglutination method, RT-PCR method, and enzyme-linked immunosorbent method. Among them, RT-PCR method and enzyme-linked immunoassay method require large-scale instruments and professionals, while hemagglutination method is simple to operate, but Human red blood cells are needed, which are expensive and not suitable for long-term storage. This study uses the principle of immune colloidal gold to develop ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983
Inventor 武玉香沈志强曲光刚唐世云孟卫芹
Owner SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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