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Preparation method and application of anti-porcine rotavirus egg yolk antibody

A pig rotavirus and egg yolk antibody technology, applied in the field of egg yolk antibody preparation, can solve the problems that piglets cannot be completely protected from infection, there is no antiviral treatment method, and the mortality rate of piglets is high, so as to reduce morbidity and mortality. , the effect of reducing RVA and mortality

Active Publication Date: 2021-12-28
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The high prevalence of RVA in pigs has caused great economic losses to the pig industry. The treatment of RVA infection can only be treated symptomatically by supplementing fluids and electrolytes, and there is no effective antiviral treatment.
RVA-specific passive maternal antibodies (local action) acquired through ingestion of colostrum (absorbed systemically and re-secreted into the intestine) are also key factors in protecting piglets, but due to the placental barrier and insufficient milk intake by piglets, through The method of immunizing sows so that piglets can obtain maternal antibodies through lactation cannot completely guarantee that piglets are free from infection
At present, there are also attenuated vaccines on the market to prevent and treat porcine rotavirus, but it takes about one month for the generation of specific antibodies after the attenuated vaccine is immunized, during which time the mortality rate of piglets infected with rotavirus is relatively high

Method used

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  • Preparation method and application of anti-porcine rotavirus egg yolk antibody
  • Preparation method and application of anti-porcine rotavirus egg yolk antibody
  • Preparation method and application of anti-porcine rotavirus egg yolk antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0035] The preparation of embodiment 1 G9P[23]RVA inactivated vaccine

[0036] (1) Cultivation and inactivation of the virus: recover the preserved porcine-derived G9P[23] strain, cultivate and collect the virus liquid, add β-propiolactone inactivator with a final concentration of 0.025% to the virus liquid, and Inactivate in a constant temperature shaking incubator at 37°C for 12 hours.

[0037] (2) Virus inactivation effect test: Inoculate the inactivated virus antigen onto MA-104 cells, place them in a constant temperature incubator at 37°C and 5% CO2 for 48-72 hours, observe CPE, and set up a negative control at the same time and the non-inactivated RVA positive control, if the positive control has typical RVA cytopathy, but the cells inoculated with the inactivated virus solution and the cells of the negative control group do not appear cytopathic, then the cell culture inoculated with the inactivated virus solution is reintroduced. After 2 generations of blind passage, ...

Embodiment 2

[0042] The preparation of embodiment 2 yolk antibody

[0043] (1) Immunization of chickens and collection of eggs

[0044]After one week of acclimatization, the laying hens were immunized with 1 mL of G9P[23]RVA inactivated vaccine through the legs and / or chest muscles, and immunized 4 times in a row with an interval of 2 weeks between each time. When it reaches 1:200 or more, it is considered qualified for immunity. Eggs are collected every day, marked, and stored at 4°C for later use.

[0045] (2) Determination of neutralizing titer of egg yolk antibody

[0046] After the second immunization, 5 eggs were randomly collected from the immunized group and the non-immunized group every 7 days, sterilized in 0.1% bromogeramine aqueous solution at 42°C for 15 minutes, taken out and dried in the air, and the egg yolk was separated with an egg white and yolk separator; Take 1 mL of the mixed egg yolk, add 9 mL of deionized water and mix again, and adjust the pH value of the egg yol...

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Abstract

The invention discloses a preparation method of an anti-porcine rotavirus egg yolk antibody. The preparation method comprises the following steps that an RVA inactivated vaccine for immunization is injected, continuously immunizing is carried out for 4 times at an interval of 2 weeks, after two immunizations, when the neutralizing titer of the antibody reaches 1: 200 or above, it is considered that the immunization is qualified, eggs are collected every day, marking is carried out, storing is carried out at 4 DEG C for later use, and the egg yolk antibody IgY is extracted. The egg yolk antibody has a certain effect on prevention and treatment of piglet RVA, can remarkably shorten the disease course, and can be used for preparing products for detecting, preventing and / or treating related diseases caused by porcine rotavirus infection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of an anti-porcine rotavirus egg yolk antibody. Background technique [0002] Group A rotavirus (Group A rotavirus, RVA) belongs to Reoviridae (Reoviridae), rotavirus (Rotavirus), is a non-enveloped double-stranded RNA virus, is an important zoonotic virus, is It is one of the important pathogens that cause diarrhea in infants and young animals worldwide, causing significant economic losses. Initially, RVA was considered to be host-specific, but now more and more epidemiological data show that RVA can be transmitted from animals to humans, or from one animal to another. [0003] Porcine group A rotavirus is an important pathogen that causes severe diarrhea in piglets. The infected piglets mainly show symptoms such as diarrhea, dehydration, vomiting, and anorexia. The high prevalence of RVA in pigs has caused great economic losses to the pig indu...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02C12N7/06G01N33/569A61K39/42A61P31/14A61P1/12A23K20/147A23K50/30C12R1/93
CPCC07K16/10C07K16/02C12N7/00G01N33/56983A61P31/14A61P1/12A23K20/147A23K50/30C07K2317/11C07K2317/76C12N2720/12363G01N2333/14G01N2469/10Y02A50/30
Inventor 张斌陈小飞汤承岳华
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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