Indirect ELISA (enzyme linked immunosorbent assay) kit based on GPV (goat poxvirus) P32 protein and preparation method

A kit and protein technology, used in biological testing, material testing and other directions, can solve the problems of frequent outbreaks of herd immunity and lack of real-time monitoring methods, achieve a good level of goat pox serum antibodies in the body, reduce blindness, and simple operation Effect

Inactive Publication Date: 2012-02-15
GUIZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the control of goat pox is mainly accomplished through vaccination. However, with the development of animal husbandry in my country and the extensiveness of livestock transactions, cross-provincial transactions occur from time to time, and the uncertainty of herd immunity in different regions has led to this Outbreaks of illness from time to time
At present, my country has not yet established a commercial goat pox serum antibody detection kit, which makes the clinical control of the disease lack a key real-time monitoring method. Therefore, it is very necessary and urgent to establish a fast and feasible goat pox serum antibody detection method

Method used

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  • Indirect ELISA (enzyme linked immunosorbent assay) kit based on GPV (goat poxvirus) P32 protein and preparation method
  • Indirect ELISA (enzyme linked immunosorbent assay) kit based on GPV (goat poxvirus) P32 protein and preparation method
  • Indirect ELISA (enzyme linked immunosorbent assay) kit based on GPV (goat poxvirus) P32 protein and preparation method

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Embodiment Construction

[0030] Embodiments of the invention:

[0031] The raw materials and formula of the iELISA kit based on GPV P32 protein are:

[0032] Goat pox standard positive serum and standard negative serum, 1:50 times diluted for use; coating buffer: Na 2 CO 3 1.59g, NaHCO 3 Dissolve 2.93g in deionized water, dilute to 1000mL, and adjust to pH 9.6; Enzyme-labeled secondary antibody: rabbit anti-goat IgG-HRP, diluted 1:2000 times for use; 1×PBST buffer: NaCl 8.0g, KCl 0.2g, KH 2 PO 4 0.2g, Na 2 HPO 4 12H 2 Dissolve O 2.9g, Tween-20 0.5mL in 800mL distilled water, adjust the pH to 7.4, and set the volume to 1000mL; blocking buffer: 5% skim milk; phosphoric acid-citric acid buffer: 5.11g citric acid monohydrate, anhydrous Na 2 HPO 4 7.3 g, dilute to 1 000 mL with deionized water; chromogenic solution: 40 mg OPD dissolved in 100 mL phosphoric acid-citric acid buffer and 12 mmol / L H 2 0 2 ; Stop solution: 1.25 mol / L H 2 SO 4 solution.

[0033] The preparation steps of the iEL...

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Abstract

The invention discloses an indirect ELISA kit based on GPV P32 protein and a preparation method. The kit comprises 50 to 200 mu L of a recombinant protein coating, 50 to 200 mu L of positive control substances, 50 to 200 mu L of negative control substances, 50 to 200 mu L of confining liquid, 50 to 200 mu L of enzyme-marked secondary antibodies, 50 to 200 mu L of a substrate solution, 50 to 200 mu L of a stopping solution and 25 to 100 mu L of washing liquid. The invention provides an effective evaluation means for immunization of goat pox vaccines on a goat farm and enables blindness in production of immunization of vaccines and production cost to be reduced.

Description

technical field [0001] The invention relates to an indirect ELISA kit and a preparation method based on GPV P32 protein. Background technique [0002] Goat pox is caused by goat pox virus ( Goat poxvirus , GPV) is an acute and severe infectious disease that seriously endangers the sheep industry. The International Veterinary Health Organization lists it as a class A infectious disease, and the Ministry of Agriculture of my country also lists it as a Therefore, my country's veterinary health system has also put forward higher requirements for the prevention of this disease. [0003] Goat pox is a disease that can be well controlled by vaccine immunization, but due to some unpredictable reasons, immune failure occurs from time to time. Serological testing is a key link in formulating and implementing goat pox control measures. Currently, commonly used serological testing methods include virus neutralization test (VNT), agar immunodiffusion test (AGID), enzyme-linked immunos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 程振涛文明周碧君岳筠王开功
Owner GUIZHOU UNIV
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