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Solid-phase competition ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit of duck tembusu virus antibody

A technology of duck tembusu virus and detection kit, which is applied in biological testing, measuring devices, material inspection products, etc. It belongs to the effect of source difference, low cost and simple operation

Inactive Publication Date: 2013-04-24
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current ELISA method for duck Tembusu virus antibody detection is indirect ELISA. This method can only be used for specific duck serum samples, and can only judge the negative or positive of serum samples. There is no standard for detecting antibody titers, so it cannot detect the tested Antibody titer of sample

Method used

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  • Solid-phase competition ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit of duck tembusu virus antibody
  • Solid-phase competition ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit of duck tembusu virus antibody
  • Solid-phase competition ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit of duck tembusu virus antibody

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Embodiment Construction

[0028] 1. Preparation of duck Tembusu virus antigen

[0029] When Vero (African green monkey kidney cells: commonly used cells for virus culture) cell growth density reaches about 80%, take the duck Tembusu virus (10 5 TCID 50 ), after melting on ice, make a 5-fold dilution with OPTI-MEM, take 2.5mL to infect 75cm 2 Incubate the cells in the cell bottle at 37°C for 1 hour, shake gently 2-3 times every 15 minutes, then discard the virus infection solution, add 10mL OPTI-MEM, and finally place at 37°C, 5% CO 2 cultured in a constant temperature incubator. After 3 days, the cells became lesioned. After the lesioned cells floated, put the cell bottle at -80°C, freeze and thaw three times, then centrifuged at 4°C and 8000rpm for 30 minutes to remove cell debris, and then put the supernatant at 4°C, Centrifuge at 35,000rpm (horizontal rotor) for 3 hours, discard the supernatant, suspend the precipitate with an appropriate amount of PBS solution, measure the concentration with B...

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Abstract

The invention discloses a solid-phase competition ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit of a duck tembusu virus antibody. The detection kit of duck tembusu virus serology disclosed by the invention comprises an ELISA plate coated with duck tembusu virus inactivated antigen, positive serum of anti-rabbit duck tembusu virus, positive control serum of anti-duck duck tembusu virus, and duck negative control serum. By adopting the kit, susceptible animal serums of duck tembusu viruses can be detected without distinguishing species; the detection kit not only can detect a field serum sample at high throughput but also can detect the antibody valence of the sample according to requirements; Simultaneously, high-end instrument equipment is unnecessary; and the solid-phase competition ELISA detection kit provided by the invention is easily grasped, and is easy to popularize and low in cost and is applied to detection of the field serum sample and laboratorial research and application.

Description

technical field [0001] The invention relates to a veterinary detection kit, specifically, the invention is a solid-phase competitive ELISA detection kit for duck Tembusu virus antibody. Background technique [0002] The duck flock disease caused by duck Tembusu virus has now formed a large-scale outbreak. Its main clinical symptoms are a sudden drop in feed intake, a rise in body temperature, green loose stools, paralysis of some ducks, depression in some ducks, and laying eggs. After the onset of the disease, the egg production of ducks dropped sharply from more than 90% to less than 10%. Autopsy showed follicle rupture and bleeding, yolk peritonitis, spleen hemorrhage, pancreas hemorrhage and necrosis, and liver enlargement and hemorrhage. The disease incidence rate is as high as 90%, and the mortality rate can reach 5%-30%. [0003] In 2010, Chinese scholars reported that Duck Tembusu virus was isolated for the first time in Hebei Province. The virus spread rapidly in so...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/96G01N33/569
Inventor 朱启运付钰广吉艳红
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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