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Detection kit for detecting infectious bovine rhinotracheitis virus antibody

A technique for rhinotracheitis virus and detection kit, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of reduced sensitivity of detection methods and difficulty in meeting the requirements of establishing ELISA for antigen purity.

Inactive Publication Date: 2017-02-15
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the prokaryotic expression product of the outer membrane glycoprotein gene of IBR virus is an insoluble protein, which requires denaturation and renaturation to obtain a purified product. After denaturation and renaturation, the spatial structure of the expression product often changes to a certain extent, resulting in The sensitivity of the detection method is reduced; in addition, the purity of the antigen obtained by the above two purification methods is difficult to meet the requirements for the establishment of ELISA

Method used

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  • Detection kit for detecting infectious bovine rhinotracheitis virus antibody
  • Detection kit for detecting infectious bovine rhinotracheitis virus antibody
  • Detection kit for detecting infectious bovine rhinotracheitis virus antibody

Examples

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Embodiment 1

[0056] The ELISA detection method of embodiment 1 anti-bovine infectious rhinotracheitis virus antibody

[0057] 1. Test method

[0058] 1. Preparation of IBRV Viral Antigen

[0059] Inoculate IBRV to MDBK cells growing in a single layer at 1% virus dose, and collect the virus when the CPE reaches 80% to 90%. Freeze and thaw repeatedly at -40°C for 3 times, centrifuge at 5000r / min (4500×g) for 30min to remove cell debris, put the supernatant into a treated dialysis bag, and then concentrate the virus solution with PEG6000. Then the concentrated virus solution was ultracentrifuged at 35000r / min (54200×g) for 2h, and finally 1 / 100 of the original cell culture was dissolved and precipitated with 0.01M PBS. After the protein concentration of the diluted product was determined, it was aliquoted and frozen at -20°C until use.

[0060] 2. Determination of the optimal coating concentration of monoclonal antibody and viral antigen

[0061] Use the square array titration method to s...

Embodiment 2

[0140] The composition of embodiment 2 test kits

[0141] An example of the composition of the kit is as follows:

[0142]

[0143] .

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Abstract

The invention provides a detection kit for detecting an infectious bovine rhinotracheitis virus antibody. The detection kit comprises an elisa plate for enveloping a monoclonal antibody and an IBRV antigen of infectious bovine rhinotracheitis virus gD protein, an IBRV positive standard serum, a negative serum, an enzyme labeling secondary antibody, a substrate developing solution and a stop solution. Infectious bovine rhinotracheitis viruses in infected MDBK cell lysates are captured in the elisa plate by using the monoclonal antibody of the infectious bovine rhinotracheitis virus gD protein, and the optimal enveloping concentrations of the monoclonal antibody and the virus antigen are determined through a square titration method. The detection kit provided by the invention has good specificity and sensitivity, is high in neutralization coincidence rate, and has a good application prospect.

Description

technical field [0001] The invention relates to antibody detection technology, in particular to a detection kit for detecting anti-bovine infectious rhinotracheitis virus antibody, and also relates to a preparation method of the kit. Background technique [0002] Infectious bovine rhinotracheitis (IBR), also known as "necrotizing rhinitis" or "red nose disease", is an exposure of cattle caused by infectious bovine rhinotracheitis virus (IBRV) STDs. Clinically, the sick cattle mainly present with respiratory symptoms such as inflammation of the respiratory tract and tracheal mucosa, dyspnea, cough, runny nose, etc. disease [1] . The disease can delay the growth of fattening cattle, reduce the milk production of dairy cows, reduce fecundity, abortion, etc., increase the culling rate and mortality of cattle herds, and bring huge economic losses to the cattle industry [2] . The virus can also invade the trigeminal ganglion and sacral ganglion of cattle. The neutralizing ant...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/535
CPCG01N33/577G01N33/535G01N33/56994G01N2333/06
Inventor 李永清许健黄秀芬叶丽娜
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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