Screening method for porcine interferon inducer

A technology of pig interferon and screening method, applied in the field of pig feed additives, can solve the problems of unclear screening purpose, low accuracy of screening results, long screening period, etc., and achieve the effects of rapid screening, strong pertinence and accurate results

Active Publication Date: 2018-06-19
HUBEI HAOHUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the screening cycle of this method is long, and the accuracy...

Method used

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  • Screening method for porcine interferon inducer
  • Screening method for porcine interferon inducer
  • Screening method for porcine interferon inducer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] The screening of embodiment 1 porcine interferon inducer

[0084] (1) In vitro culture of porcine small intestinal cell line IPEC-1

[0085] Take out the cryopreservation tube containing the porcine small intestinal cell line IPEC-1 (provided by Professor Wu Guoyao of Texas A&M University) from the liquid nitrogen, put it into a CO2 incubator to thaw, and put the porcine small intestinal cell line IPEC-1 into Petri dish (with a surface area of ​​75 cm 2 ), and add 10 times the surface area of ​​the culture medium, mix well and place in CO 2 cultured in an incubator (CO 2 The volume concentration is 5%, and the culture temperature is 37°C).

[0086] When the cells cover 90% of the surface area of ​​the culture dish, wash it twice with PBS, then add 2 mL of trypsin, and place in CO 2 After digesting in the incubator for 10 minutes, add 3 mL of complete medium to stop the digestion, then transfer all of it to a 50 mL centrifuge tube, centrifuge at 1000 rpm for 3 minute...

Embodiment 2

[0103] The further screening of embodiment 2 porcine interferon inducer

[0104] (2) Select 40 piglets of three weeks of age who are healthy and have similar body weight without specific pathogens, as test piglets, and are randomly divided into group a, group b, group c and group d (10 piglets in each group), wherein, group a The blank control group was fed with normal rations; the b group, c group and d group were the test groups, and the inducer A, inducer B and inducer C were added to the diet respectively, and the feeding time was 15 days. .

[0105] (2) On the 8th day and the 15th day of the test, the blood samples of group a, group b, group c and group d test piglets were collected respectively through the anterior vena cava, and the solid-phase sandwich ELISA was used to quantitatively detect IFN-β kit ( The Interferon betaPig ELISA Kit of Abcam Company), measure the concentration level of IFN-β in the blood sample, assay method is as follows:

[0106] ①Place the reag...

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Abstract

The invention discloses a screening method for a porcine interferon inducer. The screening method comprises the following steps: in-vitro culture of porcine small intestinal cells IPEC-1; culture of acell line IPEC-1/AcGFP1-IFN-beta for stably expressing a green fluorescent protein gene and a porcine interferon beta gene; inoculation of cells IPEC-1/AcGFP1-IFN-beta into a culture plate to form acell climbing plate; use of an inducer to be screened to treat the cells IPEC-1/AcGFP1-IFN-beta to obtain cells to be tested, observation of AcGFP1 fluorescence intensity of the cells to be tested, and screening of the porcine interferon inducer according to the AcGFP1 fluorescence intensity. The screening method for the porcine interferon inducer has the advantages of rapid screening, strong targeting and accurate results, and the selected porcine interferon inducer can be directly used as a feed additive for adding into a feed for piglets to induce production of interferon in bodies of the piglets to achieve the effect of resisting piglet diseases.

Description

technical field [0001] The invention relates to the technical field of pig feed additives, in particular to a screening method for pig interferon-inducing agents. Background technique [0002] With the advancement of science and technology and the needs of modern animal husbandry, more and more attention has been paid to the prevention and health care effects of immune enhancers. Immunity enhancers with definite characteristics, high efficiency, stability and non-toxicity will be ideal substances for preventing and treating livestock and poultry diseases in the future. . Immunopotentiators for pigs mainly include interferon (Interferon, IFN), transfer factor for pigs, immunoglobulins, oligosaccharides, astragalus polysaccharides, etc. Among them, interferon is a variety of inducers (viruses, bacteria, macromolecules, etc.) A class of proteins that induce cells to produce anti-virus, anti-tumor and immune regulation functions play a vital role in the prevention and treatment...

Claims

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Application Information

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IPC IPC(8): G01N21/64C12N15/66C12N15/85
CPCC12N15/66C12N15/85G01N21/6428G01N21/6486
Inventor 侯永清吴涛丁斌鹰易丹
Owner HUBEI HAOHUA BIOTECH
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