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Anti-TEGV (Transmissible Gastroenteritis Virus) high-immune serum and preparation method thereof

A technology of hyperimmune serum and gastroenteritis, applied in the direction of antiviral immunoglobulin, derived serum immunoglobulin, etc., can solve the problems of increased morbidity and mortality of infected pigs, weakened therapeutic effect of hyperimmune serum, etc. Good character, high antibody titer, stable virulence effect

Active Publication Date: 2014-04-09
兆丰华生物科技(南京)有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, once the clinically prevalent transmissible gastroenteritis virus mutates, the therapeutic effect of the hyperimmune serum thus prepared will be weakened, and the morbidity and mortality of infected pigs will increase

Method used

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  • Anti-TEGV (Transmissible Gastroenteritis Virus) high-immune serum and preparation method thereof
  • Anti-TEGV (Transmissible Gastroenteritis Virus) high-immune serum and preparation method thereof
  • Anti-TEGV (Transmissible Gastroenteritis Virus) high-immune serum and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Preparation of porcine transmissible gastroenteritis virus HB08 strain immunogen

[0020] Take an appropriate amount of porcine transmissible gastroenteritis-positive small intestine from a pig farm in Hebei, scrape the intestinal mucosa and contents, add PBS at a ratio of 1:5 (weight: volume), freeze and thaw repeatedly 3 times, and centrifuge to get the supernatant, 0.22 μm Filter with a filter membrane, add trypsin with a final concentration of 20 μg / ml to the filtrate, and treat at 37° C. for 1.5 hours.

[0021] Inoculate ST cells with a single layer according to the conventional method, inoculate the virus at a ratio of 10%, absorb at 37°C for 1 hour, supplement the cell maintenance solution (containing 10 μg / ml trypsin), and culture in a 37°C incubator. In this way, the cells showed slight CPE changes at the 8th passage, and obvious and stable CPE changes at the 17th passage. The cells shrank, the particles increased, and the clusters were like bunches o...

Embodiment 2

[0028] Example 2 Serum Preparation

[0029] 1) Immunization program

[0030] The first immunization: a total of 4ml intramuscular injection.

[0031] The second immunization: 14 days after the first immunization, the immunization dose is 8ml, and the injection is divided into two points.

[0032] The third immunization: 14 days after the second immunization, the immunization dose is 16ml, and injected in three points.

[0033] 2) blood collection

[0034] 15-21 days after the last immunization, (the blood was collected once, the serum titer was measured by the neutralization test method, and then bled to death, with an interval of 3-4 days), the pigs were checked for health. No feed was given for the first 12 hours, but drinking water was not restricted. The butcher's knife and the skin where the knife enters are disinfected in advance, and the shallow basin containing blood is cleaned and then steam-sterilized. Sterile bloodletting.

[0035] 3) Serum separation and prep...

Embodiment 3

[0038] Example 3 Determination of serum titer

[0039] Dilute the tested hyperimmune serum with DMEM at 1:8, 1:16, 1:32...1:2048, and add an equivalent titer of 200TCID to each dilution serum 50 / 0.1ml of TGEV HB08 strain, neutralized for 1 hour at 37°C, inoculated into 6 wells of a 96-well cell plate covered with a monolayer of ST cells, 100 μl per well, and set up 6 wells of non-neutralizing virus positive control cells and only inoculated cells Maintenance solution Negative control cells 6 wells, 37°C 5% CO 2Cultivate in an incubator for 3 to 5 days, and observe CPE every day. There should be no cytopathic changes in the cell wells of the neutralization group and the negative control group, but the cells of the virus control group should appear cytopathic. The neutralization titer of the serum was taken as the highest dilution of the serum that could protect 50% of the cells from pathological changes. As can be seen from Table 1, the prepared anti-porcine transmissible ga...

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Abstract

The invention discloses an anti-TEGV (Transmissible Gastroenteritis Virus) high-immune serum, which is prepared by separating serum from immunized pigs through taking the inactivated vaccine of a TEGV HB08 strain CGMCC (China General Microbiological Culture Collection Center) No.7807 as an immunogen. The prepared high-immune serum is excellent in character, has no bacteria, mould, mycoplasma and exogenous virus pollution, and is strong in specificity and high in safety, the cure rate of artificial infection can achieve 100%, and the neutralization titer of the serum is 29 above.

Description

technical field [0001] The invention relates to a preparation method of antiserum, in particular to an anti-porcine transmissible gastroenteritis virus hyperimmune serum and a preparation method thereof. Background technique [0002] Porcine transmissible gastroenteritis (TGE) is an important contact viral intestinal infectious disease that causes pig diarrhea. The disease is a viral disease, and there is currently no effective treatment. When pregnant sows are vaccinated, piglets can acquire resistance through passive immunity through colostrum, but with the disappearance or unevenness of maternal antibodies, the resistance of piglets gradually decreases or is different, and the resistance to infectious gastroenteritis The susceptibility also behaves differently, and once the disease occurs, it will cause huge economic losses. It is feasible to use hyperimmune serum for group therapy, and it is also a passive immunization method to enable piglets to acquire immunity and a...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/06
Inventor 侯艳红王贵华赵亚荣刘明明于萍萍满坤卢会英陈翠云
Owner 兆丰华生物科技(南京)有限公司
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