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Porcine parvovirus, vaccine composition and application thereof

A vaccine composition, parvovirus technology, applied in the directions of antiviral agents, virus/phage, virus antigen components, etc., can solve the problems of safety, immune efficacy and insufficient immune period, and achieve long immune period, good safety, The effect of high immune potency

Active Publication Date: 2013-03-13
FUJIAN AONONG BIOLOGICAL TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are various porcine parvovirus inactivated vaccines on the market and have certain immune effects, their safety, immune efficacy and immune period are still insufficient.

Method used

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  • Porcine parvovirus, vaccine composition and application thereof
  • Porcine parvovirus, vaccine composition and application thereof
  • Porcine parvovirus, vaccine composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation, cultivation and characteristic determination of porcine parvovirus

[0025] 1. Isolation of porcine parvovirus

[0026] Spleen, lung and other tissue disease materials (positive for porcine parvovirus detected by PCR) were aseptically collected from a pig farm in Nanjing, which was positive for porcine parvovirus. 1000 IU / ml, freeze-thaw 3 times, centrifuge at 12000 r / min for 5 minutes, take the supernatant, and inoculate the passaged ST cells (purchased from ATCC), the inoculation volume is 0.5ml supernatant / 25cm 3 Square flasks were cultured at 37°C for 5 days and then subcultured, during which the cytopathic changes were observed. No lesions appeared in the first generation, and after the culture was frozen and thawed three times, it was transferred to the second generation and cultured, and no lesions appeared. According to the above subculture method, it was found that the third generation of cells had cytopathic changes after 48 hours of cul...

Embodiment 2

[0058] Example 2 Preparation of Porcine Parvovirus Inactivated Vaccine

[0059] 1. ST cell seed propagation and expansion culture: Take out the frozen ST cell tube from the liquid nitrogen tank, put it in a 37°C water bath to thaw quickly, transfer the cells into a centrifuge tube filled with 10ml DMEM solution, and centrifuge at 1000rpm for 5 minutes. Discard the supernatant, suspend the cells with the growth medium, then add to the cell culture flask, at 37°C, 5% CO 2 cultivated under conditions. When the cell coverage reached 100%, the cells were digested with 0.1% trypsin-EDTA solution. Then subculture at a volume ratio of 1:3. The growth medium is DMEM solution containing 10% FCS (newborn calf serum).

[0060] 2. Virus culture: Remove the growth medium from the ST cell culture with a cell growth coverage rate of 35% to 45%. Inoculate the PPV-JS strain virus solution according to the multiplicity of infection of 0.5%, add DMEM solution containing 2% FCS, and incubate ...

Embodiment 3

[0070] Example 3 Efficacy Test of Porcine Parvovirus Inactivated Vaccine

[0071] 1. Efficacy test in guinea pigs

[0072] The inoculation objects of each batch of vaccine were grouped as follows: 6 porcine parvovirus HI antibody-negative guinea pigs weighing more than 350 g were taken, 4 of which were used as the immunization group, and 0.5 ml of vaccine was injected intramuscularly into each guinea pig; the other 2 were used as the control group and were not vaccinated.

[0073] Twenty-eight days after immunization, blood was collected from each guinea pig to detect porcine parvovirus HI antibody.

[0074] The detection method of porcine parvovirus HI antibody is as follows:

[0075] Antigen working solution preparation: According to the determined antigen HA titer, dilute porcine parvovirus PPV-JS strain culture solution into 4 HA unit antigen with PBS buffer solution to obtain the antigen working solution.

[0076] Treatment of the serum to be tested: Ta...

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Abstract

The invention provides porcine parvovirus, vaccine composition and application thereof, and belongs to the technical field of biology. The microbial preservation number of the porcine parvovirus PPV-JS strain is CGMCC (China General Microbiological Culture Collection Center) No.6605. The vaccine composition provided by the invention comprises an inactivated porcine parvovirus PPV-JS strain and veterinary pharmaceutically acceptable adjuvant. The invention also provides the application of the porcine parvovirus PPV-JS strain in preparing medicaments for preventing diseases caused by porcine parvovirus. The porcine parvovirus PPV-JS strain has excellent immunogenicity, and can be used as an activated vaccine production strain and a virus seed for inspection. The vaccine composition (porcine parvovirus inactivated vaccine) provided by the invention is good in safety, high in immune efficacy and long in immunity period, and only requires single dose, so that manpower and side effect of the vaccine to a target animal can be greatly reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a porcine parvovirus vaccine strain and application thereof. Background technique [0002] Porcine parvovirus disease is a reproductive disorder caused by porcine parvovirus (PPV). It is characterized by stillbirths, deformed fetuses, mummified fetuses, and weak piglets produced by primiparous sows, while the sows have no obvious symptoms. When sows are infected in early pregnancy, the embryonic and fetal mortality rates can be as high as 80% to 100%, and it can also cause dermatitis and diarrhea in piglets. Most primiparous sows can obtain strong immunity after infection, and even last for life. However, due to the long-term detoxification of infected pigs, the disease takes root in pigs for a long time and is difficult to remove. The infected boar's sperm cells, spermatic cord, epididymis, and accessory gonads can all carry the virus, and it is easy to pass to suscept...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/12A61P31/20C12R1/93
Inventor 张雪花张道华唐波侯继波
Owner FUJIAN AONONG BIOLOGICAL TECH GRP CO LTD
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