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Bovine enterovirus egg yolk antibody and preparation

A technology of egg yolk antibody and enterovirus, applied in the direction of antiviral agent, antibody, antiviral immunoglobulin, etc., can solve the problem of insufficient research and achieve the effect of avoiding degradation, enriching treatment methods and easy to use

Pending Publication Date: 2022-05-27
NANYANG NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the fact that there are few isolated virus strains in China, the research on them is not deep enough. Therefore, it is of great significance to isolate the virus strains, conduct in-depth research on them, and develop drugs that can prevent or treat bovine enterovirus infection.

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  • Bovine enterovirus egg yolk antibody and preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Isolation and identification of BEV virus

[0024] 1.1 Sample and sample processing

[0025] Suspected bovine enterovirus infection occurred in a cattle farm in Nanyang area, and the BEV infection rate was as high as 80% after sampling RT-PCR detection. Five fecal samples of sick cows were collected from the cattle farm. The collected fecal samples were diluted with sterilized PBS buffer (pH=7.2) at 1:4 (W / V), and then fully ground and homogenized at 8000r / v. Min centrifugation for 15 min, collect the supernatant, add 2000 U / mL penicillin-streptomycin mixture, stand at 4 °C for 4 h, filter twice with a 0.22 μm filter membrane, and store at -80 °C for later use.

[0026] Virus isolation and culture

[0027] Take 1 mL of the prepared supernatant sample, inoculate it into the MDBK cells covered with monolayer, and inoculate 1 mL of PBS as a negative control at the same time, incubate it in a 37 °C incubator for 2 h, then discard the inoculum, and wash twice wi...

Embodiment 2

[0047] Example 2. Preparation of BEV inactivated vaccine:

[0048] 2.1 Preparation of virus stock solution

[0049] The MDBK cell monolayer in good growth state was discarded from the culture medium, and the bovine enterovirus type 1 EV-E-HN1817 strain was adsorbed and inoculated at 2%. After adsorption at 37 °C for 2 hours, the maintenance solution was added to continue the culture. Daily observation and recording cytopathic conditions. Harvest when more than 80% of the cells are diseased, freeze and thaw twice, take samples for semi-finished product inspection, determine the virus titer and dilute the virus solution to 10 9.0 TCID 50 / mL, the vaccine virus stock solution is obtained, and it is stored at -80°C.

[0050] inactivated

[0051] Clarify and filter the qualified vaccine virus stock solution to remove cell debris, add 10% formaldehyde solution while stirring to make the final concentration 0.2%, inactivate at 37°C for 16h, keep stirring during this period, and...

Embodiment 3

[0060] Example 3. Preparation of BEV egg yolk antibody

[0061] 3.1 Immunization program

[0062] (1) Basic immunization: by subcutaneous injection, healthy laying hens were vaccinated with the BEV inactivated vaccine prepared in Example 2, 1.0ml per bird;

[0063] (2) Booster immunization: 14 days after the basic immunization, the BEV inactivated vaccine prepared in Example 2 was injected subcutaneously, 1.0ml each;

[0064] (3) Booster immunization: 14 days after booster immunization, the BEV inactivated vaccine prepared in Example 2 was subcutaneously injected with 1.5ml each.

[0065] (4) Maintenance immunity: After booster immunization, according to the antibody titer, booster vaccination is given every 2 to 3 months, with 1.5ml each.

[0066] High Free Egg Collection

[0067] 10 days after the booster immunization, the antibody titer in the yolk of the high-immunity egg was sampled every 5 days. When the neutralization titer was not less than 1:256, the high-immunity ...

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Abstract

Aiming at the physiological characteristics of ruminants, in order to solve the problems of digestion and inactivation of the egg yolk antibody in the rumen, the invention creatively provides two preparation modes, one is an intestinal preparation which can prevent the egg yolk antibody from passing through the rumen, and research shows that after the intestinal preparation is applied to calves, the intestinal preparation has a treatment effect on calf diarrhea caused by BEV infection, and the intestinal preparation can be used for treating the diarrhea of the calves caused by BEV infection. The effect is close to that of a traditional drug therapy, but compared with an antibiotic therapy, the method is more environment-friendly and safer; the other one is a rumen bypass preparation which can be added as a part of daily ration concentrate and is convenient to use, after the egg yolk antibody is prepared into the rumen bypass preparation, the egg yolk antibody can be prevented from being degraded in the rumen and released in the intestinal tract at a fixed point, and the treatment effect on diarrhea caused by BEV infection is achieved.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a bovine enterovirus egg yolk antibody and a preparation. Background technique: [0002] Bovine enterovirus (BEV) infection is caused by bovine enterovirus ( Bovine enterovirus, BEV ), an emerging zoonotic disease mainly characterized by gastrointestinal and respiratory symptoms. According to reports, the infection spectrum of bovine enterovirus is wide, and cattle of various breeds and ages are susceptible and have a high incidence rate. After the cattle are infected with the virus, they often show fever, cough, dyspnea, and diarrhea. In severe cases, sudden death may occur. This disease is a new infectious disease in China. BEV infection is very common in cattle, and in addition to infecting cattle, it can also infect humans, sheep and other spinal animals through oral-oral, fecal-oral contact transmission. Sick cattle infected with bovine enterovirus also carry a...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02A61K39/42A61K9/00A61K9/50A61K47/26A61P1/12A61P31/14
CPCC07K16/1009A61K9/0087A61K9/0068A61K9/5036A61K47/26A61K9/5073A61K9/5089A61K9/5015A61P1/12A61P31/14C07K2317/11A61K2039/505A61K2039/552Y02A50/30
Inventor 姚伦广刘柏麟史鸿飞刘阳坤冀君冷超粮
Owner NANYANG NORMAL UNIV
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