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Method for detecting pseudorabies virus gB protein specific antibody in pig saliva

A pseudorabies virus, specific technology, applied in the field of detection of pseudorabies virus gB protein-specific antibody in pig saliva, can solve the problems of small number of samples, low proportion of pigs, large animal stress, etc., to achieve transportation and Easy to save, save sampling time, strong resistance to the effect

Inactive Publication Date: 2014-08-20
杭州贝尔塔生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many limitations in this sampling method. On the one hand, this method of collecting blood and separating serum is time-consuming and laborious. The number of samples collected is not large, and the proportion of the pig population is not high, and the stress on animals is very large.

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  • Method for detecting pseudorabies virus gB protein specific antibody in pig saliva
  • Method for detecting pseudorabies virus gB protein specific antibody in pig saliva
  • Method for detecting pseudorabies virus gB protein specific antibody in pig saliva

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Embodiment

[0026] 1) Hang a custom-made cotton rope (6 strands, 3mm in diameter, 1m in length) in the middle of the pigs or on the fence, with the height of the bottom end at the same level as the shoulder of the animal, and loosen the hanging end of the rope, and wait for the pigs to bite After 30 minutes, recover the rope and put it in a sterile sample collection bag, place it outside the sample bag with both hands, twist the rope hard, squeeze out the saliva soaked in the rope, and collect it in a sterile 50ml centrifuge tube. Preliminary storage and transportation (at room temperature, within 6 hours, or at 4°C, within 72 hours to the laboratory). The saliva was centrifuged at 3000g for 15min, and the supernatant was taken to obtain the primary antibody of saliva, which was stored at 4°C (<7d) or -20°C (<60d) for later use, see Figure 2.

[0027] 3) Dilute the spare saliva primary antibody with PBST (1:2), and add it to the antigen-coated reaction plate (purchased from biochek in the...

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Abstract

The present invention discloses a method for detecting pseudorabies virus gB protein specific antibody in pig saliva. The method comprises: 1) adopting a cotton rope suspending method, collecting saliva, and treating to obtain a saliva first antibody; 2) adding the saliva first antibody to an antigen coated reaction plate according to the number, carrying out a 37 DEG C saturated humidity reaction for 1 h, and washing; 3) sequentially adding horseradish peroxidase-labeled goat anti-pig IgG-FC antibody to the reaction plate, carrying out a 37 DEG C saturated humidity reaction for 30 min under a dark condition, and washing; 4) sequentially adding a dimethoxybenzidine coloring agent to the reaction plate, carrying out a reaction under a dark condition for 15 min, and adding a termination solution; and 5) placing into an enzyme-labeling measuring instrument, reading the OD405 data, and determining the result. According to the present invention, the problem of single animal blood collection is avoided, and the method is suitable for large scale investigation of pseudorabies virus gB protein antibody in pig herd.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting the specific antibody of pseudorabies virus gB protein in pig saliva. Background technique [0002] Pseudorabies (Pseudorabies) is currently the most important disease that endangers the pig industry in my country, and it is endemic in most parts of the world. Pseudorabies is caused by pseudorabies virus (Pseudorabies virus, PRV), also known as Aujeszky's disease, which occurs in cattle and is called "mad itch". Pseudorabies virus is latent in the saliva and nasal secretions of infected pigs and is transmitted mainly through the nose and mouth. Pigs infected with pseudorabies usually show no symptoms, but pseudorabies virus can cause abortion, high mortality in piglets, and coughing, sneezing, high fever, constipation, lethargy, convulsions, circling, and excessive salivation in both piglets and adults. The mortality rate of piglets under 1 month old is close...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/535
CPCG01N33/54366G01N33/56994G01N2333/032
Inventor 李龙章叶恩曹洋洋
Owner 杭州贝尔塔生物技术有限公司
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