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Indirect enzyme linked immunosorbent assay (ELISA) of deer's brucellosis

An enzyme-linked immunosorbent assay, Brucella cerevisiae technology, applied in the field of veterinary medicine, can solve problems such as large gaps, and achieve the effects of accurate diagnosis, sensitivity and repeatability

Inactive Publication Date: 2012-08-01
JILIN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its specificity, sensitivity, reproducibility and the results obtained by using ELISA method to detect brucellosis antibodies in cattle and other animals are quite different.

Method used

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  • Indirect enzyme linked immunosorbent assay (ELISA) of deer's brucellosis
  • Indirect enzyme linked immunosorbent assay (ELISA) of deer's brucellosis
  • Indirect enzyme linked immunosorbent assay (ELISA) of deer's brucellosis

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0058] 1. Material preparation

[0059] (1) Material:

[0060] Brucella porcine type 2 vaccine (S 2 ), standard positive serum, standard negative serum;

[0061] (2) Preparation of enzyme-labeled antibody-rabbit anti-deer IgG-HRP:

[0062] Dissolve 5mg of horseradish peroxidase (HRP) in 0.5ml of distilled water, add freshly prepared 0.06M NaIO 4 Mix 0.5ml of aqueous solution, put it in the refrigerator at 4°C for 30min, take it out and add 0.5ml of 0.16M ethylene glycol aqueous solution, after standing at room temperature for 30min, add 1ml of aqueous solution containing 5mg of purified rabbit anti-deer IgG, mix well and put it into dialysis Put the bag in 800ml, 0.05M pH9.5 carbonate buffer and stir for 6h to combine. Then, absorb from the dialysis bag, add NaBH 4 Solution (5mg / ml) 0.2ml, placed in 4 ℃ refrigerator for 2h, the above conjugate mixture was added to an equal volume of pH7.0 saturated (NH 4 ) 2 SO 4 solution, placed in a refrigerator at 4°C for 30 minutes...

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Abstract

Indirect ELISA of deer's brucellosis belongs to the veterinary field and is used for diagnosing deer's brucellosis to improve the diagnosis level of the prior art. Biomaterials used in the assay of the invention comprise pig brucella vaccine 2 (S2) as an antigen, an enzyme labeled antibody rabbit anti-deer IgG-HRP, a standard positive serum, a standard negative serum and a serum to be assayed; and the assay comprises the following steps: coating the antigen, washing, adding the serum, adding the rabbit anti-deer IgG-HRP, adding a substrate liquid, terminating a reaction, testing by an enzyme-linked detector, adjusting to zero at 490nm, testing the OD value of each hole, and determining the result is positive when the OD value is equal to or greater than 0.32. According to the invention, the S2 is used as the antigen, the rabbit anti-deer IgG-HRP is used as the enzyme labeled antibody, and the indirect ELISA is used to diagnose the deer's brucellosis, so compared with previous diagnosis methods, the indirect ELISA of the invention has the advantages of good specificity, sensitivity and repeatability, and accurate diagnosis.

Description

technical field [0001] The invention belongs to veterinary medicine, in particular to a serum detection and diagnosis method for deer brucellosis technical background [0002] Brucellosis is an important zoonotic infectious disease worldwide caused by Brucella infection. It is extremely harmful, not only affecting people's health, hindering the development of animal husbandry, but also causing serious economic losses. [0003] Deer brucellosis is a chronic infectious disease of deer caused by Brucella. The disease mainly affects the reproductive system, causing abortion and mastitis in female deer, and orchitis and epididymitis in male deer. Since deer brucellosis was first established in 1940, deer brucellosis has occurred widely all over the world, bringing serious economic losses to the deer industry. According to foreign reports, the infection rate of deer brucellosis in some deer herds is as high as 50% to 60%, and some deer herds are even higher. Currently, there is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/535
Inventor 杜锐刘东旭时坤李建明刁燕刘晓张梦于淼
Owner JILIN AGRICULTURAL UNIV
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