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Mink parvovirus attenuated vaccine strain and its application in preparation of mink parvovirus attenuated vaccine

A mink parvovirus and attenuated vaccine technology, which is applied in antiviral agents, virus/bacteriophage, and microbial-based methods, can solve the problems of hidden dangers in the next use of vaccines, incomplete inactivation, and affecting the quality of mink fur.

Active Publication Date: 2017-03-15
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the existing mink parvovirus enteritis vaccines in the domestic market are inactivated vaccines that have not been freeze-dried. In the production process of inactivated vaccines, the inactivation is often incomplete or the use of inactivating agents (mostly formaldehyde) exceeds the standard. , which brings great hidden dangers to the next use of the vaccine
At the same time, formaldehyde is highly toxic, so the use of formaldehyde as an inactivator has certain toxicity to minks, often causing local inflammatory reactions at the inoculation site, even suppuration, ulcers, affecting the quality of mink fur and causing economic losses.
At the same time, inactivated vaccines can only mobilize the body's humoral immunity, but cannot stimulate the generation of the body's cellular immune response

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Passage weakening of mink parvovirus MEVB

[0021] 1. Passage weakening of mink parvovirus MEVB

[0022] (1) The mink parvovirus vaccine strain MEVB was inoculated on cat kidney cells CRFK by synchronous inoculation for virus amplification and culture, and the cell lesion was observed every day. When the cell lesion reached about 80%, it could be frozen and thawed repeatedly. The virus was harvested using the method described above and labeled MEVB-F1 (passage 1).

[0023] (2) The first-generation virus MEVB-F1 was inoculated and harvested according to the method described above, and the virus was labeled as MEVB-F2 (second generation). Using the same method, the virus was passaged to passage 61, labeled as MEVB-F61.

[0024] (3) When the mink parvovirus MEVB was continuously passaged to the 10th generation (MEVB-F10), the virus was cloned and plaque-purified. The method is: dilute the virus to be purified 10 times to 10 -7 , take 10 -3 ~10 -7 0.1ml of v...

Embodiment 2

[0033] The preparation of embodiment 2 mink parvovirus enteritis live vaccine

[0034](1) Vaccine strain MEVB-F61 was inoculated on CRFK cells synchronously according to 0.1%-2% of the culture volume, and cultured in a 35°C spinner. Methods Harvest virus liquid. Five batches of vaccines were prepared according to the above method, and the virus content and purity were tested. The results showed that the viral titers (TCID) of the five batches of vaccines 50 )≥10 6.5 / ml, indicating that the vaccine strain MEVB-F61 has good production performance and is suitable for large-scale industrial production.

[0035] (2) Preparation of lyoprotectant

[0036] Dissolving gelatin, hydrolyzed milk protein and sucrose in water, according to the mass percentage, the final concentration of gelatin is 2-7%, the final concentration of hydrolyzed milk protein is 10-25%, and the final concentration of sucrose is 15-30%. Obtained after autoclaving at 116°C for 30 minutes.

[0037] (3) Mix th...

Embodiment 3

[0039] Example 3 Mink Parvovirus Enteritis Live Vaccine Safety Test

[0040] Get 3 batches of qualified mink parvovirus enteritis live vaccines (prepared by the method of embodiment 2), hindlimb muscle inoculation antibody negative mink, carry out single dose respectively (10 5.5 / head portion), single-dose repeated and over-dose (10 times) inoculation, while non-vaccinated mink was set as a control. After the vaccination, the body temperature, appetite, mental state and other physiological indicators of the minks were observed for 14 consecutive days, and the intestinal lesions of the vaccinated minks were observed by autopsy. The test results showed that after single-dose, repeated single-dose and over-dose vaccination, all the physiological indexes of the minks were normal, without any clinical symptoms, and there was no pathological change in the intestinal tract of the vaccinated mink. It shows that mink parvovirus enteritis live vaccine is safe for mink.

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Abstract

The invention discloses a mink parvovirus attenuated vaccine strain and its application in preparing the mink parvovirus attenuated vaccine. The mink parvovirus attenuated vaccine strain is named MEVB-F61, and is preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, and the preservation number is: CGMCC NO.9560. Tests have shown that the vaccine strain is safe and effective when used as a vaccine, and can effectively prevent the occurrence of mink parvovirus enteritis, and after 5 consecutive generations of animal inoculation experiments, it has been proved that the vaccine strain has no virulence reversion. Therefore, the invention will have broad application prospects in the field of preventing and treating mink parvovirus enteritis.

Description

technical field [0001] The invention relates to mink parvovirus attenuated vaccine strain and its application, and the invention belongs to the technical field of veterinary biological products Background technique [0002] Mink parvovirus enteritis is an acute and highly contagious infectious disease caused by parvovirus in the family Parvoviridae. Together with canine distemper and Aleutian disease in mink, it is called the three major diseases of mink farming. The disease was first reported by Schofield in 1949 and found in Canada. It was isolated and identified by Wills in 1952 and named as mink enteritis virus. Currently, mink parvovirus enteritis widely exists in various mink raising countries in the world. Minks of different ages and varieties are susceptible, but especially young minks and young minks aged 50 to 60 days are the most susceptible. The incidence rate of young minks is over 70%, and the mortality rate is as high as 90%. It is about 30%, and the mortalit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/23A61P31/20C12R1/93
Inventor 闫喜军胡博柴秀丽张海玲张蕾赵建军白雪刘昊
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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