Porcine rotavirus G4-G5-G9 type trivalent inactivated vaccine as well as preparation method and application thereof

The technology of porcine rotavirus and inactivated vaccine is applied in the field of porcine rotavirus G4\G5\G9 trivalent inactivated vaccine and its preparation, which can solve the problems of dead pig breeding industry economy, piglet disease, loss and the like, To achieve the effect of good immunogenicity, high safety and convenient operation

Active Publication Date: 2022-08-05
WUHAN KEQIAN BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Viral infections caused by porcine rotavirus are common in pigs whether alone or cross-infected, resulting in sickness or even death of piglets, which has caused huge economic losses to the pig farming industry
At present, there is no effective drug to treat the disease caused by porcine rotavirus infection, so it is particularly important to prevent porcine rotavirus infection, and vaccination can effectively prevent and control the outbreak of the disease

Method used

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  • Porcine rotavirus G4-G5-G9 type trivalent inactivated vaccine as well as preparation method and application thereof
  • Porcine rotavirus G4-G5-G9 type trivalent inactivated vaccine as well as preparation method and application thereof
  • Porcine rotavirus G4-G5-G9 type trivalent inactivated vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Isolation, identification, cloning, purification and biological characteristics of porcine rotavirus

[0042] 1.1 Isolation and identification of viruses

[0043] The disease material was obtained from the small intestine of pigs with clinical symptoms of diarrhea in large-scale pig farms in my country. The positive small intestine tissue was taken, cut into pieces with scissors, added with 10.0ml of PBS, ground with a tissue grinder or mortar, and then transferred to a 15ml centrifuge tube. Repeat freezing and thawing 3 times below -70°C, take four 1.5ml centrifuge tubes, add 1.0ml small intestine tissue fluid to each centrifuge tube, centrifuge at 12000r / min for 5 minutes, filter and sterilize with 0.22μm microporous membrane, and inoculate MA104 cells that have grown into a monolayer (drain the cell culture medium before inoculation and wash three times with serum-free DMEM medium), adsorb at 37°C for 1 hour, aspirate the inoculum, add 5ml of cell maintenance...

Embodiment 2

[0085] Example 2 Preparation and inspection of porcine rotavirus G4\G5\G9 trivalent inactivated vaccine

[0086] A preparation method of porcine rotavirus G4\G5\G9 trivalent inactivated vaccine, comprising the following steps:

[0087] 1 Culture technology of porcine rotavirus (G9-CH09, G5-KQ06 and G4-HN01 strains) virus liquid on PK-15 cells

[0088] In order to facilitate the industrial production of porcine rotavirus, the culture process of suspension PK-15 cells in the shake flask was determined by exploring the passage density and shaking speed of the suspension type PK-15 cells in the shake flask: 1.0× 10 6 Dilute passages at an initial density of cells / ml at 37°C, 130r / min, 5%CO 2 After 72 hours of culture in a horizontal shaker, the cell density can reach 6.0 to 6.5 × 10 6 When the porcine rotavirus was inoculated in a shake flask, the cell density, trypsin concentration, inoculation amount, inoculation time and other process conditions were explored, and the porcin...

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Abstract

The invention provides a porcine rotavirus G4-G5-G9 type trivalent inactivated vaccine as well as a preparation method and application thereof, three genotype strains of current PoRV epidemic are adopted as seed viruses, are inactivated by BEI, and are emulsified with an adjuvant ISA201VG to prepare the PoRV trivalent inactivated vaccine. The trivalent inactivated vaccine prepared by the research has the characteristics of high immune protection rate, good stability, high yield, safety and reliability, and can be used for emergency vaccination when a PoRV epidemic situation outbreaks.

Description

technical field [0001] The invention belongs to the field of biotechnology and discloses a porcine rotavirus G4\G5\G9 trivalent inactivated vaccine and a preparation method and application thereof. Background technique [0002] Porcine rotavirus (RV) is a double-stranded RNA virus without an envelope, belonging to the genus Rotavirus of the family Reoviridae. A disease that causes piglets with vomiting, anorexia, dehydration, diarrhea, and acid-base balance disorders as the main symptoms. The morbidity of piglets less than 4 weeks old can reach or exceed 80%, which can cause 7%-20% mortality. [0003] Rotaviruses have now identified 11 proteins, namely inner capsid proteins (VP1, VP2, VP3 and VP6), outer capsid proteins (VP7 and VP4) and nonstructural proteins (NSP1, NSP2, NS34, NS35 and NS28). Rotavirus has different serogroups and serotypes, and is divided into 7 serogroups (types) (A-G) and two subgroups (I and II) with different antigenicity according to VP6; The main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/295A61K39/15A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/14C12N2720/12321C12N2720/12334A61K2039/5252A61K2039/70A61K2039/552A61K2039/54Y02A50/30
Inventor 徐高原周明光张华伟朱娴静罗修鑫陈映瑾刘慧
Owner WUHAN KEQIAN BIOLOGY CO LTD
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