Virus clone purification strain for Peste des petits ruminants live vaccine production, and applications thereof

A technology for the production of Peste des petits ruminants and vaccines, which is applied in the direction of veterinary vaccines, antisense single-stranded RNA viruses, viruses, etc., and can solve the problems of secondary infection of other pathogens, difficult to deal with, and decreased immunity of animals

Active Publication Date: 2017-06-13
CHINA INST OF VETERINARY DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Peste des petits ruminants virus has an immunosuppressive effect. After infection, it will lead to a decline in the immunity of animals, causing secondary infections of other pathogens, making the epidemic more complicated and difficult to deal with

Method used

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  • Virus clone purification strain for Peste des petits ruminants live vaccine production, and applications thereof
  • Virus clone purification strain for Peste des petits ruminants live vaccine production, and applications thereof
  • Virus clone purification strain for Peste des petits ruminants live vaccine production, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] ——Clone and Purify the Production Virus of Peste des Petits Ruminants Live Vaccine

[0033] After the applicant introduced the vaccine strain from the Peste des Petits Ruminants Reference Laboratory, he found that the cell lesions were irregular, the lesion appeared for more than 96 hours and was not typical, and the virus content was low, especially in the process of virus breeding. During the neutralization test, the lesions were very irregular. In order to select strains with strong reproductive ability, high virus titer and suitable for vaccine production on Vero cells, the introduced strains were cloned and purified, and the clones obtained by cloning and purification were named It is Pester des petits ruminants virus (PPRV) 75 / 1 Clone 9 strains, which were sent to the Institute of Microbiology, Chinese Academy of Sciences, No. 1, Courtyard 1, Beichen West Road, Chaoyang District, Beijing on December 15, 2016 General Microorganisms Collection Center of China Commit...

Embodiment 2

[0066] ——Preparation of seed batches for the production of live vaccines against Peste des petits ruminants

[0067] The effective way to prevent PPR is to vaccinate against PPR vaccine. In order to develop the live vaccine of Peste des petits ruminants, 2 batches of basic seeds for production of live vaccines of Peste des petits ruminants and 1 batch of production seeds were specially prepared and systematically tested. The research results are reported as follows:

[0068] 1. Virus reproduction

[0069] After hydrating the PPR 75 / 1Clone9 freeze-dried virus seed with serum-free MEM, inoculate a Vero cell monolayer with 1% of the cell maintenance solution, and culture it at 37°C for about 120 hours after inoculation. Freeze and thaw the bottle at -20°C for 3 times, harvest the virus liquid, and pass it to the 145th generation in this way; add the 141st and 145th generation virus liquids into the freeze-drying protective agent at a ratio of 1:1, divide into ampoules, each 1ml...

Embodiment 3

[0104] ——Preparation of live vaccine against Peste des petits ruminants

[0105] (1) Virus seeds for production The basic seed virus for production of PPRV 75 / 1Clone 9 strains was prepared and provided by the China Veterinary Drug Control Institute.

[0106] (2) Vero cells for production: preserved and provided by China Veterinary Drug Administration.

[0107] (3) Preparation of Peste des petits ruminants virus liquid After the PPRV virus seeds were hydrated with serum-free MEM, Vero cell suspensions with different cell concentrations were inoculated synchronously at 0.1%, 0.5%, 1%, and 2%, and the cell suspension concentrations were respectively Adjusted to 200,000 to 300,000 / ml, 500,000 to 600,000 / ml. After mixing, pack the cell flasks separately, place them in a 37°C incubator, observe the cell lesions every day, and record the time when more than 70% of the lesions appear. When the lesions reach more than 70%, move the cell bottles to -20°C and freeze-thaw 3 times repeatedl...

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Abstract

The present invention relates to a virus clone purification strain for Peste des petits ruminants live vaccine production, and applications thereof. According to the present invention, an introduced Peste des petits ruminants virus attenuated PPRV 75 / 1 strain is cloned and purified to obtain a PPRV 75 / 1Clone9 strain, ie., the virus clone purification strain, wherein the virus titer is increased after the cloning; the virus titer is decreased by not more than 0.3 titer during the storage and the use of the Peste des petits ruminants live vaccine prepared by using the virus clone purification strain; the safety and the immunizing efficacy are higher than the OIE standard; and the immunity protective test results show that the immunizing 10<2>TCID50 protection rate is 96.7%, and the immunizing 10<3>TCID50 protection rate is 100%.

Description

technical field [0001] The invention relates to the establishment and application of virus clone and purified strain for production of live vaccine of Peste des petits ruminants. It belongs to the field of veterinary biological products. Background technique [0002] Peste des petits ruminants (PPR), also known as Pseudo-rinderpest, is an acute, virulent, contact disease caused by Peste des petits ruminants virus (PPRV). Infectious disease; OIE classifies it as a class A infectious disease, the incidence rate of susceptible groups is as high as 100%, and the mortality rate reaches 100% when the outbreak is severe. At present, there is no effective treatment for the disease, and it is mainly controlled by early diagnosis and vaccine immunization. Since the disease was first reported in Côte d'Ivoire in the 1840s, it has been prevalent in most African countries, the Arabian Peninsula, Israel, Syria, Iraq, Jordan, Turkey and other Middle East regions, as well as the Indian Pe...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/155A61P31/14C12R1/93
CPCA61K39/12A61K2039/552C12N7/00C12N2760/18421C12N2760/18434
Inventor 薛青红印春生孙淼张广川支海兵
Owner CHINA INST OF VETERINARY DRUG CONTROL
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