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Porcine rotavirus strain, vaccine composition, preparation method and application thereof

The technology of porcine rotavirus and vaccine composition is applied in the field of porcine rotavirus strain and vaccine composition and preparation field, and can solve the difficulty of separation of porcine rotavirus, the lack of effective drugs for porcine rotavirus disease, and the inability of pigs to rotavirus. Prevention and control of rotavirus disease

Active Publication Date: 2021-04-27
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease is widespread worldwide, resulting in slow growth or increased mortality in piglets, causing huge economic losses to the pig industry
[0004] Currently, there is no effective drug for porcine rotavirus disease, conventional treatment methods are poor, and vaccine is the main measure to prevent and control the disease
However, the isolation of porcine rotavirus is difficult, and the virus titer of the isolated virus strain is low, which is unfavorable for the later stage of cultivation and proliferation, resulting in low immune efficacy of the prepared vaccine composition, which cannot effectively treat porcine rotavirus disease. prevention and control

Method used

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  • Porcine rotavirus strain, vaccine composition, preparation method and application thereof
  • Porcine rotavirus strain, vaccine composition, preparation method and application thereof
  • Porcine rotavirus strain, vaccine composition, preparation method and application thereof

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Embodiment approach

[0038] As an embodiment of the present invention, the inactivated whole virus antigen is the whole virus culture solution obtained by propagating the porcine rotavirus strain on cells and inactivated by an inactivating agent. As an embodiment of the present invention, the inactivating agent includes but not limited to β-propiolactone BPL, binary ethyleneimine BEI, formalin, formaldehyde, N-acetylethyleneimine AEI, polyhexamethylene hydrochloride Methylguanidine PHMG.

[0039] As a preferred embodiment of the present invention, in the vaccine composition of the present invention, the subunit antigen VP7 protein content of the porcine rotavirus HN03 strain or its culture is 25-150 μg / ml.

[0040] As a preferred embodiment of the present invention, in the vaccine composition of the present invention, the content of the subunit antigen VP7 protein of the porcine rotavirus HN03 strain or its culture is 50 μg / ml.

[0041] The subunit antigen VP7 protein of the porcine rotavirus HN0...

Embodiment 1

[0057] The isolation of embodiment 1 porcine rotavirus HN03 strain

[0058] 1. Sample screening and processing

[0059] Feces and small intestinal contents were collected from 2-3 day-old piglets with the disease in various places, and all were positive for porcine rotavirus by test strips. After diluting the feces and small intestine contents 4 times with PBS (0.02mol / L, pH value 7.4), centrifuge at 8000rpm for 10 minutes, take the supernatant and filter it with a 0.22μm filter, then store it in aliquots, and name them respectively as sample 1 and sample 1. 2. Sample 3, Sample 4, Sample 5, Sample 6, Sample 7 and Sample 8.

[0060] 2. Isolation of porcine rotavirus

[0061] 2.1 Separation method of the present invention Separation: Separation is carried out on MA104 cells. After the MA104 cells were subcultured (the cell culture medium was α-MEM+5% FBS), 1.2-1.5 million cells were inoculated into 25T cell flasks. After the cells grow into a monolayer, treat the treated sma...

Embodiment 2

[0069] The identification of embodiment 2 porcine rotavirus HN03 strain

[0070] 1 Detection of porcine rotavirus HN03 strain

[0071] Design a pair of primers for the VP7 gene, which can be detected universally by RT-PCR method. The sequences of the primers used are as follows:

[0072] F: 5'-CCCCGGTATTGAATATACCACAGT-3'

[0073] R: 5'-TTTCTGTTGGCCACCCTTTAGT-3'

[0074] According to the instructions of the nucleic acid extraction kit (Geneaid company), RNA was extracted from the porcine rotavirus HN03 strain virus liquid, and reverse transcription was carried out according to the instructions of EasyScript First-Strand cDNA Synthesis SuperMix (TransGen company). The reaction system was (20 μl) : 2×ES Reaction Buffer 10μl, ES RT / RI Enzyme Mix 1μl, random primer 1μl, RNase-free water 4μl, RNA template 4μl. The reaction conditions are: 10 minutes at 25°C, 30 minutes at 42°C, and 5 minutes at 85°C. PCR was performed using cDNA as a template, and the system was as follows (25 μ...

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Abstract

The invention relates to a porcine rotavirus strain HN03 strain. The strain has good immunogenicity, and the vaccine composition prepared from its inactivated antigen and subunit antigen can have a good protective effect on pigs. The preparation of the invention contains pigs. The method for the vaccine composition of rotavirus inactivated antigen can cultivate the virus that produces high droplet toxicity, and the antigen content is high, thereby ensuring the immune efficacy of the vaccine.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a porcine rotavirus strain and a vaccine composition, preparation method and application thereof. Background technique [0002] Rotavirus (RV) is a member of the Reoviridae family and the genus Rotavirus, and is the main pathogen that causes viral diarrhea in infants and various young animals. Piglets, calves, sheep, goats, foals, chickens, turkeys, and other poultry can be infected with rotavirus. [0003] Porcine Rotavirus (PRoV) mainly causes diarrhea in piglets, accompanied by clinical symptoms such as anorexia, vomiting, and dehydration. The disease is mainly transmitted through feces and can infect pigs of various ages and genders. The infection rate of piglets at 3 to 5 weeks is relatively high. After piglets are infected with porcine rotavirus, the duration of shedding in feces is about 1 to 14 days, and It is often co-infected with en...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/15A61P31/14
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/55511C12N7/00C12N2720/12321C12N2720/12334
Inventor 田克恭汪志艳孙进忠张许科
Owner PU LIKE BIO ENG
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