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Detection kit and detection method for porcine pseudorabies gB antibody

A porcine pseudorabies and antibody detection technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of long detection time, poor repeatability, and difficulty in quantification, and achieve the effect of short reaction time, large surface area, and improved repeatability

Active Publication Date: 2018-07-20
上海鸣捷生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problems of long detection time, difficult quantification, poor repeatability, and difficult automation in the existing veterinary detection technology, the purpose of the present invention is to provide a swine pseudorabies gB antibody detection kit, which can quantitatively detect swine pseudorabies gB Antibody content

Method used

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  • Detection kit and detection method for porcine pseudorabies gB antibody
  • Detection kit and detection method for porcine pseudorabies gB antibody
  • Detection kit and detection method for porcine pseudorabies gB antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0098] A kit for detecting porcine pseudorabies gB antibody, comprising a magnetic suspension coated with porcine pseudorabies gB protein, alkaline phosphatase-labeled porcine pseudorabies gB antibody solution, diluent, calibrator, quality control, cleaning solution, Luminescent substrate.

[0099] Preparation of magnetic suspension coated with porcine pseudorabies gB protein:

[0100] (1) Take 1 mL of magnetic particles (purchased from JSR Corporation, Japan, item number: Magnosphere TM MS300 / Caboxyl) solution with a concentration of 10 mg / mL, washed twice with 0.1 mol / L 2-morpholineethanesulfonic acid (MES) buffer solution with a pH of 5.0, and finally suspended in 1 mL of 0.1 mol / L pH 5.0 In MES buffer;

[0101] (2) 50 μg of purified porcine pseudorabies gB protein (purchased from Hangzhou Yiminuo Biotechnology Co., Ltd.) was added;

[0102] (3) Weigh 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), dissolve it with 0.1mol / L MES buffer solution with pH 5.0, so that ...

Embodiment 2

[0168] A test kit for porcine pseudorabies gB antibody detection, comprising a magnetic suspension coated with porcine pseudorabies gB protein, acridinium ester-labeled goat anti-pig IgG antibody (purchased from Beijing Soleibao Technology Co., Ltd.) solution, diluent, Calibrator, Quality Control, Washing Solution, Primary Luminescent Substrate and Second Luminescent Substrate.

[0169] Preparation of magnetic suspension coated with porcine pseudorabies gB protein:

[0170] (1) Take 1mL of the solution containing magnetic particles with a concentration of 10mg / mL, wash it twice with 0.1mol / L boric acid buffer solution with a pH of 9.5, and finally suspend in 1mL of a 0.1mol / L boric acid buffer solution with a pH of 9.5;

[0171] (2) Add 60 μg of purified porcine pseudorabies gB antigen, vortex and mix;

[0172] (3) Add 0.5 mL of 0.1 mol / L boric acid buffer (containing 3 mol / L ammonium sulfate) with a pH of 9.5, and shake and react at 37°C for 20 hours;

[0173] (4) Add 0.5mL...

Embodiment 3

[0222] A kit for detecting porcine pseudorabies gB antibody, comprising magnetic suspension coated with porcine pseudorabies gB protein, horseradish peroxidase-labeled porcine pseudorabies gB antigen solution, diluent, calibrator, quality control, cleaning solution, the first luminescent substrate and the second luminescent substrate.

[0223] Preparation of magnetic suspension coated with porcine pseudorabies gB protein:

[0224] (1) Take 1mL of the solution containing magnetic particles, the concentration is 10mg / mL, wash twice with 0.01mol / L phosphate (PBS) buffer solution with pH 7.4, and finally suspend in 1mL 0.01mol / L PBS with pH 7.4 in the buffer;

[0225] (2) Add 0.1 mL of 25% (v / v) glutaraldehyde solution, shake and react at 37°C for 2 hours;

[0226] (3) Wash 3 times with 1mL 0.01mol / L PBS buffer solution with pH 7.4;

[0227] (4) Adding 50 μg of purified porcine pseudorabies gB recombinant antigen, shaking and reacting at 37° C. for 20 hours;

[0228] (5) Add 0...

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Abstract

The invention discloses a detection kit and a detection method for a porcine pseudorabies gB antibody. The kit detects the content of the antibody in a sample to be detected through a specific reaction of a magnetic particle-coated antigen, a luminescent marker-labeled protein and the antibody in the sample to be detected. The kit realizes the quantitative detection of the porcine pseudorabies gBantibody, and has the advantages of high sensitivity, wide detection range, fastness, good repeatability, and realization of fully automatic operation and high-throughput detection.

Description

technical field [0001] The invention relates to the technical field of porcine pseudorabies gB antibody detection, in particular to a porcine pseudorabies gB antibody detection kit and a detection method using the kit for detection. Background technique [0002] Pseudorabies (PR) is an acute infectious disease caused by Pseudorabies Virus (PRV). Adult pigs are often invisible infections, and pregnant sows feel abortion, stillbirth and mummified fetuses; congenital infection or early infection of pseudorabies virus causes neurological symptoms and high mortality in suckling piglets; for nursery pigs and fattening pigs, pseudorabies virus is Important causative agent of respiratory disease syndrome (PRDC), causing respiratory symptoms and growth retardation. The disease widely exists in our country and is one of the important infectious diseases. Once an animal is infected, the virus can remain in a state of latent infection for a long time in the body, and can survive in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/532
CPCG01N33/532G01N33/54326G01N33/56983G01N2333/032Y02A50/30
Inventor 宋启超左青山纪良心刘聪李炎晖
Owner 上海鸣捷生物科技有限公司
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