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Porcine pseudorabies virus (PRV) variant PRV-ZJ01 and application thereof

A technology of PRV-ZJ01 and porcine pseudorabies, applied in antiviral agents, viruses/phages, medical preparations containing active ingredients, etc., to achieve good immunogenicity and safety

Active Publication Date: 2014-03-12
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In order to solve the problem that the epidemic situation existing in the above prior art cannot be effectively controlled, the invention provides a kind of porcine pseudorabies virus variant strain PRV-ZJ01

Method used

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  • Porcine pseudorabies virus (PRV) variant PRV-ZJ01 and application thereof
  • Porcine pseudorabies virus (PRV) variant PRV-ZJ01 and application thereof
  • Porcine pseudorabies virus (PRV) variant PRV-ZJ01 and application thereof

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Embodiment 1

[0032] 1. Virus isolation and PCR identification

[0033] The collected brain tissue of diseased pigs was shredded and mixed (the weight-to-volume ratio of tissue to sterilized PBS was 1:10), centrifuged at 12,000 rpm for 10 min, and the supernatant was filtered through a 0.22 μm filter for virus isolation. Inoculate 1mL of the filtered supernatant of the disease material into BHK-21 cells that have just grown into a single layer, incubate at 37°C for 1h, discard the liquid in the culture flask, and add 2% DMEM in a solution containing 5% CO 2 Cells were cultured in a 37°C cell culture incubator, and the cells were observed for lesions every day. After 70% of the cells had lesions, they were frozen and thawed three times, and BHK21 cells were inoculated. Virus plaques were purified 3 times, and the gE gene was amplified by PRV PCR method. Reaction system: 1 U AccuPrime TM pfx DNA Polymerase with 2.5μL 10×AccuPrime TM pfx Mix (Invitrogen), upstream and downstream primers...

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Abstract

The invention relates to the technical field of porcine pseudorabies viruses (PRVs) and in particular relates to a porcine PRV variant PRV-ZJ01 with collection number of CGMCCNo.8170 and an application of the porcine PRV variant PRV-ZJ01 in preparation of vaccines. The porcine PRV variant PRV-ZJ01 has the beneficial effects that a water-soluble inactivated vaccine is prepared by adopting a PRV-ZJ01 variant virus solution and is subjected to a swine immune protection test with live vaccines of Bartha-K61, Bucharest and HB-98 strains and the results show that the inactivated vaccine of the ZJ01 strain has relatively high safety and has the immune protection efficiency obviously higher than that of immunity groups of the live vaccines of the Bartha-K61, Bucharest and HB-98 strains, and the live vaccines of the Bartha-K61, Bucharest and HB-98 strains can not provide full protection for the ZJ01 very virulent strain; the inactivated vaccine of ZJ01 has relatively good immune protection effects on the PRV variant and the traditional strains; infected with 10<6.0>TCID50 (Tissue culture infectious dose 50) / ml nasal drops of the PRV-ZJ01 variant, all the 85-day-old non-immune swine can become ill and die; results prove that the virulence of the virus strain is obviously enhanced, the antigenicity is varied and the virus strain has relatively good immunogenicity after being inactivated and can be used for research and development of the vaccine of the virus strain and the diagnostic methods.

Description

technical field [0001] The invention relates to the technical field of porcine pseudorabies virus, in particular to a variant strain of porcine pseudorabies virus PRV-ZJ01, and also relates to the application of the variant strain of porcine pseudorabies virus PRV-ZJ01. Background technique [0002] Pseudorabies virus (PRV) is a member of the herpesviridae herpesvirus A subfamily. It can cause central nervous system disorders in newborn piglets, respiratory symptoms in growing pigs, and can cause abortion, stillbirth and weak piglets in pregnant sows. The United States and some European countries have declared the eradication and eradication of the disease. The gene-deficient attenuated vaccine is widely used in my country to effectively control and gradually purify it. However, since the end of 2011, many pig farms immunized against the disease in my country have successively experienced piglet vomiting and neurological symptoms, growing pigs have respiratory symptoms, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/245A61P31/22C12R1/93
Inventor 姜平顾真庆白娟
Owner NANJING AGRICULTURAL UNIVERSITY
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