Detection kit and detection method for porcine pseudorabies gE antibody

A porcine pseudorabies and antibody detection technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of long detection time, difficulty in automation, and low sensitivity, and achieve short reaction time, wide detection range, and good repeatability Effect

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

[0005] In order to solve the problems of long detection time, low sensitivity, poor repeatability and difficulty in automation in the existing veterinary detection technology, the purpose of the present invention is to provide a swine pseudorabies gE antibody detection kit, which can quickly and sensitively detect pig Pseudorabies gE antibody

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] A kit for detecting porcine pseudorabies gE antibody, comprising a magnetic suspension coated with porcine pseudorabies gE protein, alkaline phosphatase-labeled porcine pseudorabies gE antibody solution, diluent, calibrator, quality control, cleaning solution, Luminescent substrate.

[0096] Preparation of magnetic suspension coated with porcine pseudorabies gE protein:

[0097] (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;

[0098] (2) Add 80 μg of purified full-length porcine pseudorabies gE protein (purchased from Hangzhou Yiminuo Biotechnology Co., Ltd.);

[0099] (3) Weigh 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), dissolve it with 0.1mol / L MES buffer solution with pH 5.0, so...

Embodiment 2

[0158] A test kit for porcine pseudorabies gE antibody detection, comprising a magnetic suspension coated with porcine pseudorabies gE 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.

[0159] Preparation of magnetic suspension coated with porcine pseudorabies gE protein:

[0160] (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;

[0161] (2) Add 90 μg of purified porcine pseudorabies gE antigen, vortex and mix;

[0162] (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;

[0163] (4) Add 0.5mL...

Embodiment 3

[0208] A kit for detecting porcine pseudorabies gE antibody, comprising a magnetic suspension coated with porcine pseudorabies gE protein, horseradish peroxidase-labeled porcine pseudorabies gE antigen solution, diluent, calibrator, quality control, and cleaning solution, the first luminescent substrate and the second luminescent substrate.

[0209] Preparation of magnetic suspension coated with porcine pseudorabies gE protein:

[0210] (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;

[0211] (2) Add 0.1 mL of 25% (v / v) glutaraldehyde solution, shake and react at 37°C for 2 hours;

[0212] (3) Wash 3 times with 1mL 0.01mol / L PBS buffer solution with pH 7.4;

[0213] (4) Adding 30 μg of purified porcine pseudorabies gE recombinant antigen, shaking and reacting at 37° C. for 20 hours;

[0214] (5)...

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Abstract

The invention discloses a detection kit and a detection method for a porcine pseudorabies gE 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 gEantibody, 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 gE antibody detection, in particular to a porcine pseudorabies gE 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): G01N33/577G01N33/569G01N33/543
CPCG01N33/54326G01N33/56983G01N33/577G01N2333/032G01N2446/90Y02A50/30
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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