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

Abstract

The present invention discloses a method for detecting pseudorabies virus gE 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 18-25 DEG C saturated humidity reaction for 1 h, and washing; 3) sequentially adding horseradish peroxidase-labeled goat anti-pseudorabies virus gE protein IgG to the reaction plate, carrying out a 18-25 DEG C saturated humidity reaction for 30 min under a dark condition, and washing; 4) mixing TMB coloring agent solutions A and B according to a ratio of 1:1, sequentially adding 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 OD650 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 gE 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 gE 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...

Examples

Embodiment

[0025] 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). Centrifuge the saliva at 3000g for 15min, take the supernatant to obtain the primary antibody of saliva, and store it at 4°C ( figure 2 .

[0026] 2) Dilute the spare saliva primary antibody with PBST (1:1), and add it to the antigen-coated reaction plate according to the number (purchased from IDEXX Company in the United States, product nu...

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for detecting the specific antibody of pseudorabies virus gE 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...

Examples

Embodiment

[0025] 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). Centrifuge the saliva at 3000g for 15min, take the supernatant to obtain the primary antibody of saliva, and store it at 4°C ( figure 2 .

[0026] 2) Dilute the spare saliva primary antibody with PBST (1:1), and add it to the antigen-coated reaction plate according to the number (purchased from IDEXX Company in the United States, product nu...

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