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Raccoon dog parvovirus enteritis virus LN strain, raccoon dog parvovirus enteritis inactivated vaccine and preparation method of vaccine

A technology for toxic enteritis and inactivated vaccines, applied in biochemical equipment and methods, antiviral agents, viruses/bacteriophages, etc., can solve the problems of time-consuming, laborious, unstable quality, and large batch-to-batch differences, and achieve stable virus proliferation, Good immunogenicity and shortened production cycle

Inactive Publication Date: 2019-05-14
JL TEYAN BIOLOGICAL TECH LIMITED LIABILITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the unsatisfactory immune effect of most of the inactivated mink parvovirus enteritis vaccines or canine parvovirus attenuated vaccines used to immunize raccoon dogs, and the process of cultivating vaccines by rotating bottles is complicated, long-term, time-consuming and labor-intensive, and of poor quality. For the problems of stability, large batch-to-batch variation, and high side effects, the invention provides a raccoon parvovirus LN strain, an inactivated raccoon parvovirus vaccine and a preparation method thereof

Method used

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  • Raccoon dog parvovirus enteritis virus LN strain, raccoon dog parvovirus enteritis inactivated vaccine and preparation method of vaccine
  • Raccoon dog parvovirus enteritis virus LN strain, raccoon dog parvovirus enteritis inactivated vaccine and preparation method of vaccine
  • Raccoon dog parvovirus enteritis virus LN strain, raccoon dog parvovirus enteritis inactivated vaccine and preparation method of vaccine

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preparation example Construction

[0059] The present invention adopts the method of continuous passage of sensitive F81 adherent cells and the full suspension process, separates, clones and purifies raccoon parvovirus from the intestinal tract of raccoon dogs with disease on the market, and obtains a raccoon dog strain that is safe to raccoon dogs, has good immunogenicity and stable virus proliferation Parvovirus enteritis vaccine strain. The inactivated vaccine of raccoon parvovirus enteritis was prepared by F81-TY full suspension cell culture technology combined with improved β-propiolactone inactivation, emulsification and seedling mixing. A kind of preparation method of raccoon parvovirus enteritis inactivated vaccine of the present invention mainly comprises the following steps:

[0060] Step 1. Resuscitation of Suspension Cells

[0061] Recovery of F81-TY strain cells;

[0062] Step 2. Suspension cell passage

[0063] The F81-TY strain cells revived in step 1 are expanded and cultured with low-serum s...

Embodiment 1

[0089] Example 1 Isolation and Identification of Raccoon Parvoviral Enteritis Virus LN Strain

[0090] 1. Virus isolation

[0091] (1) Disease material treatment

[0092] The collected disease materials (in Luannan area, Hebei Province) were diluted with an equal amount of normal saline, thoroughly mixed and oscillated, centrifuged at 3000rpm for 30min, the supernatant was taken, and stored in a -20°C refrigerator for later use.

[0093] (2) Virus isolation

[0094] Dilute the treated disease material with MEM nutrient solution at a volume ratio of 1:2, inoculate in a bottle of adherent cat kidney cells (F81) simultaneously, culture at 37°C for 24 hours, and replace the nutrient solution with cell maintenance solution (containing 2 % serum MEM culture fluid), observe the cytopathic changes day by day, culture for 4-5 days to collect the poison, continue blind passage for 4 generations, detect the HA characteristics of the culture, and store the positive ones for future use. ...

Embodiment 2

[0099] Preparation of embodiment 2 raccoon parvovirus inactivated vaccine (suspension process)

[0100] 1. Suspension cell recovery

[0101] Take the F81-TY strain low-serum full-suspension cells cryopreserved in liquid nitrogen, thaw quickly in a 37°C water bath and inoculate in a shake flask containing 400 mL of low-serum suspension culture medium with a serum concentration of 1-5%, and place at 37°C. ℃, 60rpm, 5% (v / v) CO 2 Incubate on a shaker for 72 hours until the cell density reaches 8.0×10 5 cells / mL and cell viability ≥ 95%, the revived F81-TY strain cells were obtained.

[0102] 2. Suspension cell passage

[0103] The resuscitated F81-TY strain cells obtained in step 1 were scaled up with low-serum suspension culture medium to obtain F81-TY strain cell culture medium.

[0104] Its specific operation project is as follows:

[0105] (1) Inoculate the revived F81-TY strain cells obtained in step 1 in a shaker flask filled with low-serum suspension culture medium at...

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Abstract

The invention provides a raccoon dog parvovirus enteritis virus LN strain, a raccoon dog parvovirus enteritis inactivated vaccine and a preparation method of the vaccine, and belongs to the field of veterinary biological products. The raccoon dog parvovirus enteritis virus LN strain is preserved in the China General Microbiological Culture Collection Center on December 7th, 2018, and the preservation number is CGMCC NO.16627. The effective active ingredient of the raccoon dog parvovirus enteritis inactivated vaccine is the raccoon dog parvovirus enteritis virus LN strain. The raccoon dog parvovirus enteritis inactivated vaccine is prepared through a cell full suspension technology, and can resist strong virus attacks of the raccoon dog parvovirus, the protection rate is 91.7% or above, raccoon dog viral enteritis can be effectively prevented, and the effective immune period is 6 months. The problems that when existing vaccines are used for immunizing raccoon dogs, the effect is not ideal, the process and period for vaccine spinner cultivation are long, the quality is not stable, the batch-to-batch difference is large and side reactions are high are solved.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a raccoon parvovirus LN strain, an inactivated vaccine of raccoon parvovirus and a preparation method thereof. Background technique [0002] Raccoon parvovirus enteritis is an acute and highly contagious infectious disease, which has become one of the important infectious diseases that seriously endanger the raccoon breeding industry in recent years. Raccoon parvovirus enteritis occurred for the first time in Heilongjiang Province of my country in 1984. Clinically, the body temperature rises above 40 degrees; mental depression, vomiting, loss of appetite or exhaustion, and increased thirst; severe diarrhea, discharge of watery stools mixed with blood and mucus or loose stools mixed with intestinal mucosa. The emergence of cast and so on. Raccoon parvovirus enteritis has become more and more serious year after year. At present, there is no vacc...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/23A61K39/39A61P31/20
Inventor 任飞倪佳王春霞许皓吕文雪赵江平李云松王萃瑜张亭亭杨升刘建荣
Owner JL TEYAN BIOLOGICAL TECH LIMITED LIABILITY
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