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Preparation and use method of canine coronavirus chemiluminescence detection reagent

A chemiluminescence detection and coronavirus technology, which is applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems of no technical solution, cumbersome operation, and inapplicability, and achieve the effect of rapid detection, high sensitivity, and good accuracy

Pending Publication Date: 2021-11-02
广州艺得诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pet disease diagnosis is different from human disease diagnosis. Pet hospitals are far smaller than human hospitals. Conventional chemiluminescence detectors are too bulky and cumbersome to operate in pet hospitals. With the portability and rapidity of chemiluminescence detection instruments , and the development of easy operation, chemiluminescence detection technology will have great potential in the rapid screening market of pet infectious diseases
[0004] However, at present, no practical and specific technical solutions have been researched, and the chemiluminescent immunoassay method can be used for early screening and detection of canine coronavirus enteritis.

Method used

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  • Preparation and use method of canine coronavirus chemiluminescence detection reagent
  • Preparation and use method of canine coronavirus chemiluminescence detection reagent
  • Preparation and use method of canine coronavirus chemiluminescence detection reagent

Examples

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Embodiment 1

[0036] Embodiment 1: A kind of preparation method of canine coronavirus chemiluminescence detection reagent, described canine coronavirus chemiluminescence detection reagent comprises magnetic microsphere-G19 conjugate and G15-acridine ester conjugate, and specific preparation process includes as follows step:

[0037] Preparation of magnetic microsphere-G19 conjugates: use EDC and NHS to activate the magnetic microspheres, add canine coronavirus antibody G19 for oscillating coupling for 4 hours, block the magnetic microspheres with blocking solution for 2 hours, wash the magnetic microspheres twice in the magnetic stand, and then place them in the Preserve in the magnetic microsphere buffer, and set aside to obtain the magnetic microsphere-G19 conjugate;

[0038] Preparation of G15-acridine ester conjugates: centrifuge and concentrate canine coronavirus antibody G15 and acridinium ester to remove impurities, mix G15 and acridinium ester thoroughly according to the mass ratio ...

Embodiment 2

[0044] Example 2: Using the same preparation method as in Example 1 above, a chemiluminescent detection reagent for canine coronavirus was obtained.

[0045] Then dilute a series of standard substances with different concentrations, and then perform luminosity RLU detection on the standard substances with different concentrations of canine coronavirus one by one by using the following canine coronavirus chemiluminescence detection reagent, the specific operation steps are as follows:

[0046] Take dog vomit or feces samples, dilute appropriately with sample diluent, add 50 μL of diluted standards of different concentrations, 30 μL of magnetic microspheres-G19 conjugates, 40 μL of G15-acridine ester conjugates in a transparent reaction tube The reaction tube was incubated with shaking at room temperature for 10 minutes, and then washed 3 times under the action of a magnetic field. The reaction tube entered the detection darkroom, and the substrate excitation solution A and the b...

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Abstract

The invention discloses a preparation method of a canine coronavirus chemiluminescence detection reagent, and belongs to the technical field of biological detection preparations. The method comprises the following steps: preparing a magnetic microspheron-G19 conjugate: activating magnetic microspheres by adopting EDC and NHS, adding a canine coronavirus antibody G19, oscillating and coupling the material, and storing the material in a magnetic microsphere buffer solution for later use; preparing a G15-acridinium ester conjugate: fully and uniformly mixing G15 and acridinium ester, carrying out labeling coupling, carrying out oscillation labeling, sealing, purifying, and storing in a luminous reagent buffer solution for later use. The method is used to obtain the luminosity RLU of the detected sample. According to the preparation and use method of the chemiluminescence detection reagent, the blank of domestic canine coronavirus chemiluminescence detection reagents is filled, and qualitative, quantitative and rapid detection of canine coronavirus antigens is truly realized.

Description

technical field [0001] The invention relates to the technical field of biological detection preparations, in particular to a preparation method and application of a detection reagent for canine coronavirus enteritis. Background technique [0002] Canine coronavirus enteritis is a viral infectious disease source caused by canine coronavirus (CCV), which can cause gastroenteritis symptoms of varying degrees in dogs, characterized by frequent vomiting, diarrhea, depression, anorexia and other symptoms. The disease can occur throughout the year, and it occurs frequently in winter. Sick dogs are the main source of infection. Dogs can infect through the respiratory tract, digestive tract, feces and pollutants. Once the disease occurs, the littermates are difficult to control and can cause infection, and the mortality rate of puppies is relatively high. Canine coronavirus enteritis is one of the most common viral infectious diseases in dogs. In view of the serious harm of canine ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/532G01N33/531G01N33/543
CPCG01N33/56983G01N33/532G01N33/531G01N33/54326G01N2333/165G01N2469/10
Inventor 李来庆陈翠翠梁焕坤宁波郭桂铃昝洁赖宏锐
Owner 广州艺得诺生物科技有限公司
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