A new goose parvovirus, inactivated vaccine and its application

Abstract

The present invention provides a novel goose parvovirus, an inactivated vaccine and an application thereof, and belongs to the technical field of veterinary biological products. The present invention isolates and obtains a novel goose parvovirus, and poultry will develop neurological symptoms 2-3 days after being infected by the strain. The present invention inactivates the novel goose parvovirus, and the prepared novel goose parvovirus inactivated vaccine has good safety, can provide effective immune protection for ducks, and has a good prospect for commercial development. When preparing the novel goose parvovirus inactivated vaccine, the present invention first inoculates the novel goose parvovirus into DEF cells in the S phase to obtain a virus liquid, and on this basis, selects a suitable formula ratio to prepare an oil phase and an aqueous phase, and performs a fully emulsified preparation method, which improves the safety and stability of the prepared vaccine while improving the efficiency of the inactivated vaccine preparation.

Description

Technical Field

[0001] The invention relates to the technical field of veterinary biological products, and in particular to a novel goose parvovirus, an inactivated vaccine and applications thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Waterfowl parvovirus belongs to the Parvoviridae family, Anseriformes order, Dependoparvovirus anseriform 1, and is a non-enveloped single-stranded DNA virus. The virus exists in an icosahedral symmetrical structure formed by a capsid with a diameter of about 20-22 nm, and usually uses the hairpin structure at the end of its own genome to complete autonomous viral DNA replication. Novel goose parvovirus (NGPV) mainly attacks young ducklings. The disease is widely prevalent in northern my country, causing a significant drop in the duck slaughter rate and causing serious damage.

[0004] The new goose parvovirus infection mainly occurs in ducklings under 3 weeks old and is highly contagious. Infected ducklings often show symptoms of "short beak and long tongue" and feather loss on the back. The feather loss symptoms appear about 7-10 days after infection, while the "short beak and long tongue" symptoms will not appear until about 15-20 days after infection. Therefore, farmers cannot immediately take preventive measures such as isolation of uninfected ducks after being infected with the new goose parvovirus. Ducklings infected with the new goose parvovirus are shorter than normal ducks when they grow up, and the elimination rate of disabled ducks is as high as 80%, and the market rate is low, causing serious economic losses to meat duck farmers. Therefore, it is of great significance to strengthen the isolation and identification of the new goose parvovirus strains, detect the symptoms of the new goose parvovirus infection as early as possible, and carry out effective immune intervention in the uninfected ducks in a timely manner to prevent the disease.

[0005] For the prevention and control of diseases caused by the new goose parvovirus infection, in addition to timely physical isolation measures, the development of effective vaccines is crucial. However, the proliferation of the new goose parvovirus in in vitro culture, such as embryo or cell culture, is complex and unpredictable, which makes the development of the new goose parvovirus vaccine difficult. At present, the prevention and control of the new goose parvovirus on the market all use goose plague vaccines for prevention, which has poor prevention effects, and conventional antiviral and antibacterial treatments are ineffective. The inactivated vaccine is simple to produce, highly safe and easy to store. Therefore, it is extremely urgent to develop a safe and efficient inactivated vaccine that can prevent the new goose parvovirus. Summary of the invention

[0006] Based on the above prior art, the purpose of the present invention is to provide a novel goose parvovirus, an inactivated vaccine and applications thereof.

[0007] To achieve the above object, the present invention adopts the following technical solution:

[0008] In the first aspect of the present invention, a novel goose parvovirus is provided. The novel goose parvovirus has been deposited in the General Microbiological Center of China Microorganism Culture Collection Committee (referred to as CGMCC, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing) on ​​October 24, 2024, and the deposit name is: NGPV-NMG21, and the classification name is: novel goose parvovirus ( Anser anser novus parvovirus ), the deposit number is: CGMCC No.46205.

[0009] The complete genome nucleotide sequence of the novel goose parvovirus is shown in SEQ ID NO:1.

[0010] The novel goose parvovirus (NGPV-NMG21) provided by the present invention is a variant strain of the existing novel goose parvovirus. Poultry infected by it will show neurological symptoms in the early stage. In addition, poultry that died after infection were examined, and it was found that poultry infected by this strain also showed obvious gallbladder enlargement. The above symptoms have never appeared in previous novel goose parvovirus infections.

[0011] The neurological symptoms are specifically manifested as depression and limb incoordination when walking in poultry 2-3 days after infection with the strain.

[0012] Preferably, the poultry is ducklings or goslings.

[0013] The second aspect of the present invention provides the use of the novel goose parvovirus in preparing a vaccine for preventing and / or treating the novel goose parvovirus.

[0014] By investigating the cell tropism of the novel goose parvovirus provided by the present invention, it was found that the strain has a wide range of cell tropism, can infect a variety of animal cells and can proliferate in large quantities in a short period of time. Therefore, the novel goose parvovirus (NGPV-NMG21) provided by the present invention can be used as an excellent vaccine strain for the preparation of vaccines.

[0015] The term "cell tropism" refers to the ability to infect and / or replicate in one or more cell lines or tissue cells.

[0016] The vaccine is any one of an inactivated vaccine, a live attenuated vaccine, a subunit vaccine or a genetically engineered vaccine.

[0017] The third aspect of the present invention provides a novel inactivated goose parvovirus vaccine, which comprises the inactivated novel goose parvovirus.

[0018] The novel inactivated goose parvovirus vaccine further comprises a pharmaceutically acceptable adjuvant.

[0019] The third aspect of the present invention provides a method for preparing the novel goose parvovirus inactivated vaccine, comprising the following steps:

[0020] (1) The novel goose parvovirus was inoculated into DEF cells in the S phase to obtain a virus solution, and formaldehyde was added to the virus solution at a final concentration of 0.2%, and the virus solution was inactivated by stirring at 37°C for 16-18 hours to obtain an inactivated antigen solution;

[0021] (2) Add Tween-80 to the inactivated antigen solution, shake and mix until Tween-80 is completely dissolved to obtain an aqueous phase, mix white oil, aluminum stearate and Span-80 evenly and sterilize to obtain an oil phase;

[0022] (3) The oil phase and the water phase were mixed at a volume ratio of 2:1 at 6000 rpm for 10 min, and then emulsified at 8000 r / min for 20 min to prepare a new goose parvovirus inactivated vaccine.

[0023] TCID of the novel goose parvovirus in the virus solution described in step (1) 50 For 10 -7.39 / 0.2 mL.

[0024] The volume ratio of the inactivated antigen solution to the added Tween-80 in step (2) is (24-49):1.

[0025] The beneficial effects of the present invention include:

[0026] 1. The present invention has isolated a new type of goose parvovirus. Poultry will develop neurological symptoms 2-3 days after being infected with the strain. In addition, poultry that died after infection were examined and found to have obvious gallbladder enlargement. The strain has a wide range of cell tropism and can be used as an excellent vaccine strain for vaccine preparation. It has been deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on October 24, 2024, with the deposit number: CGMCC No.46205. The nucleotide sequence of its whole genome is shown in SEQ ID NO:1.

[0027] 2. The new goose parvovirus provided by the present invention is inactivated to prepare a new goose parvovirus inactivated vaccine with good safety, without any local and systemic adverse reactions caused by the vaccine, and all indicators detected are stable and effective. The immune effect of the vaccine is evaluated by the immune challenge method, and the results also show that the inactivated vaccine prepared by the present invention can provide effective immune protection for ducks and has a good prospect for commercial development.

[0028] 3. When preparing a novel goose parvovirus inactivated vaccine, the present invention first inoculates the novel goose parvovirus into DEF cells that have been in the S phase of cell division for a long time to obtain virus liquid. Since the DEF cells in the S phase have strong division ability, the virus strain is inoculated into the DEF cells in the S phase, and the replication ability of the virus strain is stronger. On this basis, a suitable formula ratio is selected to prepare the oil phase and the water phase, and a preparation method of fully emulsifying is performed, which improves the preparation efficiency of the inactivated vaccine and improves the safety and stability of the prepared vaccine. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. The embodiments of the present invention are described in detail below in conjunction with the accompanying drawings, wherein:

[0030] Figure 1 This is the pathological change image of DEF virus cultured for 96 hours.

[0031] Figure 2 This is a whole genome nucleotide homology comparison of the new goose parvovirus NGPV-NMG21 and other NGPV wild strains in GenBank.

[0032] Figure 3 This is a comparison chart of the amino acid homology of the VP1 protein of the new goose parvovirus NGPV-NMG21 and other NGPV wild strains in GenBank.

[0033] Figure 4 This is a picture of the gallbladder tissue of a duckling. Figure 4 A in the middle is a picture of the gallbladder tissue of a healthy duckling. Figure 4 Figure B is a picture of the gallbladder tissue of a duckling infected with the new goose parvovirus NMG21 strain.

[0034] Figure 5 This is a diagram of the meningeal tissue of a duckling. Figure 5 A in the middle is a picture of the brain meningeal tissue of a healthy duckling. Figure 5 Figure B is a picture of the meningeal tissue obtained by shaving a duckling infected with the new goose parvovirus NMG21 strain.

[0035] Figure 6Determination of virus growth curve.

[0036] Figure 7 To test the purity of the virus. DETAILED DESCRIPTION

[0037] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0038] As mentioned above, the infection of the new goose parvovirus will cause serious economic losses to meat duck farmers. The symptoms of feather loss in ducks infected with the new goose parvovirus strains appear about 7-10 days after infection, and the symptoms of "short beak and long tongue" will not appear until about 15-20 days after infection, so farmers cannot immediately isolate and prevent the uninfected ducks. In addition, there is currently no effective vaccine specifically for the new goose parvovirus on the market. Therefore, it is of great significance to strengthen the isolation and identification of the new goose parvovirus strains, develop a safe and efficient inactivated vaccine that can prevent the new goose parvovirus, detect the symptoms of the new goose parvovirus infection early, and carry out effective immune intervention in the uninfected ducks in a timely manner to prevent the disease.

[0039] Based on this, the present invention provides a new goose parvovirus, an inactivated vaccine and its application. In 2021, the inventors of this application discovered a case of infection with a new goose parvovirus in Inner Mongolia. Unlike in the past, in addition to the classic short beak, long tongue and feather shedding characteristics of the infected ducks, neurological symptoms also appeared in the early stage. The autopsy found that in addition to massive bleeding in tissues and organs throughout the body, the gallbladder of the infected ducks was abnormally swollen, and there was a slight fibrinous embolism at the junction of the jejunum and ileum, accompanied by a sharp swelling of the bursa of Fabricius. After 28 days of continuous observation, the final infection mortality rate of the duck flock reached 40%, exceeding the mortality rate of the traditional new goose parvovirus (10% ~ 20%). The overall rate of disabled ducks in the duck flock reached more than 70%. The diseased material was ground and inoculated into duck embryos. After three blind generations, sequencing determined that the infectious pathogen was a new type of goose parvovirus, which was deposited in the General Microbiology Center of China Culture Collection Administration on October 24, 2024, with the deposit number: CGMCC No.46205.

[0040] The whole genome of the novel goose parvovirus was sequenced. The nucleotide sequence of the whole genome of the novel goose parvovirus is shown in SEQ ID NO: 1, which is as follows:

[0041]

[0042] Furthermore, the homology between the new goose parvovirus and other NGPV wild strains in GenBank was analyzed, and it was found that the whole genome nucleotide homology of the new goose parvovirus and other NGPV wild strains in GenBank was between 98.0-99.7%, and the structural protein amino acid homology was between 97.6-99.8%.

[0043] The inventors of the present application named the above-mentioned new goose parvovirus as NMG21 strain, and prepared an inactivated vaccine for preventing the new goose parvovirus with the inactivated NMG21 strain. When preparing the inactivated vaccine, the NMG21 strain was first inoculated into DEF cells in the S phase to obtain the virus liquid. Since the DEF cells in the S phase have strong division ability, the replication ability of the strain is stronger when the strain is inoculated into the DEF cells in the S phase. On this basis, a suitable formula ratio is selected to prepare the oil phase and the water phase, and a fully emulsified preparation method is performed, which improves the efficiency of the inactivated vaccine preparation while improving the safety and stability of the vaccine preparation.

[0044] The novel goose parvovirus inactivated vaccine prepared by the present invention has good safety. The vaccinated ducks did not show any local and systemic adverse reactions caused by the vaccine. The immune effect of the vaccine was evaluated by the immune challenge method. The results showed that the inactivated vaccine prepared by the present invention can provide effective immune protection for ducks and has a good prospect for commercial development.

[0045] The following detailed description is for illustrative purposes only and is intended to provide further explanation of the present application, rather than to limit the scope of the present invention.

[0046] Example 1: Isolation and identification of a novel goose parvovirus strain NMG21

[0047] (1) Autopsy the spleen tissue of the diseased ducklings in Inner Mongolia, add sterile saline to the liver and spleen at a ratio of 1:5 (W / V) for homogenization, freeze and thaw the homogenate three times, centrifuge at 6000 rpm for 15 min, and take the supernatant for use. Add 8000 U / mL of double antibiotics (penicillin and streptomycin) to the supernatant and place it at 4℃ overnight. Take the supernatant and filter bacteria and other substances with a filter to obtain the original virus mixture.

[0048] (2) DEF cells that have been in the S phase for a long time were selected for virus isolation. DEF cells were prepared as follows: 11-day-old Cherry Valley duck embryos were taken, the eggshells were disinfected with alcohol, and the eggshells were removed along the edge of the air chamber. Sterile tweezers were used to clamp the neck of the embryo and place it in a sterile glass culture dish. The head, limbs and internal organs were removed with scissors. The embryo was transferred to a small beaker, repeatedly rinsed with PBS reagent, and then the embryo was cut into chyme. A sterile Pasteur pipette was used to place the chyme embryo in a 50 mL sterile centrifuge tube, 4 volumes of EDTA-trypsin were added, and the tube was placed in a 37°C water bath for 10 min. DMEM high-glucose medium supplemented with FBS was added to the trypsin-treated embryo, and the tissue blocks were blown away repeatedly with a Pasteur pipette to disperse the cells evenly. The suspension was filtered through 4 layers of gauze to remove unprocessed broken tissue pieces, and the cells were counted using a red blood cell counting plate. The cells were then dispensed into 6-well cell culture dishes and cultured at 37°C and 5% CO2.

[0049] During the culture process, it was found that under low serum culture conditions, a large number of DEF cells showed slow growth due to low serum content, which can keep the cells in the cell division period for as long as possible, thus maximizing the reproduction of the new goose parvovirus. Cyclin A, a gene specifically expressed in the S phase of the cell cycle, was used to quantitatively measure DEF cells under different serum concentrations. At the same time, based on the comparison of cell growth status, it was finally determined that under the DMEM culture conditions containing 3% FBS, the cells can continue to exist in the S phase in the first 36 hours of culture while maintaining a good growth trend after virus infection, and the highest virus load can be obtained at 120 hours. Excessive serum concentration will greatly shorten the S phase, resulting in a significant reduction in the virus content or even the inability to propagate the virus. However, too low serum concentration seriously affects the proliferation and resistance of cells, and ultimately leads to premature cell death.

[0050] (3) The original virus mixture was inoculated into DEF cells under low serum concentration for passage. After the third passage, the virus liquid with cytopathic effect was isolated. The virus liquid was inoculated into high-glucose DMEM medium containing 3% FBS to obtain a stable strain. The whole genome sequencing result of the strain is shown in SEQ ID NO:1. The homology of the whole genome sequencing result with other NGPV wild strains in GenBank is between 98.5% and 99.6% ( Figure 2 ), VP1 protein amino acid homology is between 97.6-99.2% ( Figure 3 ), indicating that the isolated strain is a new type of goose parvovirus, which is named NMG21 strain, and the strain is biologically preserved. The preservation information is as follows:

[0051] Strain name: NGPV-NMG21;

[0052] Classification and naming: New goose parvovirus ( Anser anser novus parvovirus );

[0053] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0054] Abbreviation of depository institution: CGMCC;

[0055] Address of the depository institution: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;

[0056] Deposit date: October 24, 2024;

[0057] Deposit number: CGMCC No.46205.

[0058] Example 2: Observation of the phenotype of ducks infected with the new goose parvovirus NMG21 strain

[0059] Ten one-day-old Cherry Valley ducks without NGPV maternal antibody interference were selected, and the third-generation cell culture of the new goose parvovirus NMG21 strain was injected intramuscularly into the ducklings. The injection volume was 0.2mL 1000TCID50 / duck. After the injection, the ducklings were observed daily. The experimental records are as follows:

[0060] Two days after the inoculation, two ducklings in the flock showed signs of limb incoordination when walking and were in a depressed state of mind. Three days after the inoculation, all four ducklings showed signs of limb incoordination when walking. Seven days after the inoculation, three of the ten ducklings showed feather loss on their backs. Eight days after the inoculation, one duckling died. In addition to the common lesions of the new goose parvovirus, the autopsy showed severe swelling of the gallbladder tissue of the dead duck ( Figure 4 ), meningeal hemorrhage ( Figure 5 ). Ten days after the inoculation, the remaining nine ducklings all had feathers falling off their backs. Fifteen days after the inoculation, three of the remaining nine ducklings had tongue ulcers and exposed tongues. Twenty days after the inoculation, the remaining nine ducklings all had tongue ulcers and exposed tongues, and the beaks of the diseased ducklings were significantly shorter than those of the healthy ducklings.

[0061] The experimental results showed that ducklings infected with the new goose parvovirus NMG21 strain will develop neurological symptoms due to meningeal hemorrhage in the early stage, and the dead ducks will show obvious gallbladder enlargement, which has never occurred in previous new goose parvovirus infections.

[0062] Example 3: Cell tropism detection of the novel goose parvovirus NMG21 strain

[0063] In order to determine the cell tropism of the novel goose parvovirus NMG21 strain, the present study used the cytopathic effect observation method to detect the proliferation of the novel goose parvovirus NMG21 strain in different cells such as duck embryo fibroblasts and duck embryo hepatocytes, and the virus growth curve of the NMG21 strain was prepared in duck embryo fibroblasts ( Figure 6 ), as follows:

[0064] (1) Test method:

[0065] Use prepared duck embryo fibroblasts and duck embryo hepatocytes to determine the virus TCID 50 Determination. Prepare cells according to the preparation method of DEF cells described in step (2) of Example 1, and transfer the cells to a 96-well plate for culture. When the cells grow to a density of 10%, inoculate the virus solution diluted tenfold into the cells of the 96-well plate, set 8 replicates for each dilution, and inoculate 200 uL of the virus dilution solution in each well. Place the inoculated cells in a 37°C incubator and continue to culture for 96 hours. Observe the cells every day for lesions and record the positive and negative wells of each dilution.

[0066] Use the prepared duck embryo fibroblasts to draw the virus growth curve. The prepared cells were transferred to a 6-well plate for culture. When the cells grew to 10% density, 500 uL of the virus stock solution was inoculated. At 6, 12, 24, 36, 48, 72 and 96 hours after inoculation, the cells in different wells were collected separately, and the cells were repeatedly frozen and thawed to fully release the virus solution into the cell supernatant. The virus solution was extracted and the number of virus copies was determined using qPCR.

[0067] (2) Test results

[0068] The results of virus content of the novel goose parvovirus NMG21 strain propagated on different cells are shown in Table 1.

[0069] Table 1: Virus content of the novel goose parvovirus NMG21 strain propagated on different cells

[0070]

[0071] Purification and cultivation of novel goose parvovirus is a difficult point in virus and vaccine research, and whether a vaccine strain that is suitable for in vitro cell culture can be obtained is the key to vaccine development and production. Different from the novel goose parvovirus reported in the prior art, the novel goose parvovirus NMG21 strain of the present invention has a wide range of cell tropism, can infect a variety of animal cells, and can produce a large number of proliferations in a short period of time. Therefore, the novel goose parvovirus NMG21 strain of the present invention can be used as an excellent vaccine strain for vaccine preparation.

[0072] Example 4: Preparation of a novel goose parvovirus inactivated vaccine

[0073] 1. Preparation of venom for production

[0074] (1) The novel goose parvovirus NMG21 strain isolated in Example 1 was inoculated into DEF cells in the well-growing S phase to proliferate the virus in large quantities. The cells and supernatant were collected and repeatedly frozen and thawed three times at -80°C to completely lyse the cells so that the viruses present inside the cells could be fully released into the cell supernatant. The cell precipitate was removed by centrifugation, and the supernatant was collected to obtain the virus solution. The TCID50 titer of the virus solution was 10 -7.39 / 0.2 mL.

[0075] (2) Use PCR to detect whether the virus solution obtained in step (1) contains other common viruses. The detection items include avian influenza virus (AIV), novel duck reovirus (NDRV), duck circovirus (DuCV), and purify the seed virus. The test results are shown in Figure 7 .

[0076] (3) Take the virus liquid that has passed the bacterial test and mix it with formaldehyde with a final concentration of 0.2% in a volume ratio of 1:200, and stir and inactivate it at 37°C for 16 h to obtain the inactivated antigen liquid.

[0077] (4) Preparation of oil phase: Mix No. 10 medicinal white oil and Span-80 in a volume ratio of 94:6, add 2 g of aluminum stearate, stir and mix well, and sterilize at high temperature and high pressure to obtain the oil phase.

[0078] (5) Preparation of aqueous phase: Add 4% of the volume of Tween-80 to the inactivated antigen solution in step (3), shake and mix to completely dissolve the Tween-80, and obtain an aqueous phase.

[0079] (6) Emulsification: The oil phase and the water phase are mixed in a ratio of 2:1. The mixing process is as follows: add 2 parts of the oil phase into a tissue homogenizer in an ultra-clean bench, and slowly add 1 part of the water phase while stirring continuously. After all the water phase is added, mix at 6000 rpm for 10 minutes, and then emulsify at 8000 r / min for 20 minutes. The mixture is divided into portions for use, and the inactivated vaccine is prepared.

[0080] Example 5: Quality inspection of inactivated vaccines

[0081] The inactivated vaccine prepared in Example 4 was subjected to quality inspections including dosage form, centrifugal stability, viscosity, sterility and shelf life. For specific methods, refer to the "Pharmacopoeia of Veterinary People's Republic of China" (2020 edition).

[0082] The results showed that the dosage form of the inactivated vaccine prepared by the present invention was water-in-oil (W / O); the centrifugal stability, viscosity and sterility tests met the requirements of the "Pharmacopoeia of Veterinary People's Republic of China" (2020 edition).

[0083] Example 6: Vaccine safety test

[0084] Thirty one-day-old ducklings (Cherry Valley ducks) were selected and randomly divided into two groups, 15 in each group. Group 1 was the test group, and the new goose parvovirus inactivated vaccine qualified by the quality inspection of Example 5 was injected intramuscularly in the chest, 0.5 mL / duck. Group 2 was the control group, and the sterilized white oil adjuvant was injected intramuscularly in the chest, 0.5 mL / duck. After vaccination, the mental state of each group was continuously monitored, as well as whether local inflammatory reactions such as redness, swelling, heat and pain occurred at the injection site. After 2 weeks, the test animals were dissected to evaluate the vaccine absorption at the injection site.

[0085] Results: Neither the experimental group nor the control group showed obvious mental depression. During the observation period, the growth and development of each group were normal. The autopsy of the experimental group showed that the vaccine was well absorbed at the injection site, and there was no inflammatory reaction such as tissue necrosis. The results showed that the inactivated vaccine was safe and harmless and had no effect on animal growth.

[0086] Example 7: Testing the protective properties of vaccines

[0087] Thirty one-day-old ducklings (Cherry Valley ducks) were selected and randomly divided into two groups, 15 in each group. Group 1 was the experimental group, and the prepared new goose parvovirus inactivated vaccine was injected intramuscularly in the chest, 0.5 mL / duck. Group 2 was the control group, and the sterilized white oil adjuvant was injected intramuscularly in the chest, 0.5 mL / duck. Seven days after immunization, 0.2 mL of the new goose parvovirus NMG21 strain virus liquid was injected into the leg muscles of the two groups of ducklings, and they were isolated and raised in groups to observe the mental state and mortality of the two groups of ducklings.

[0088] Results: The immunized groups were in good mental state, with normal individual development, no neurological symptoms, feather loss, short beak and long tongue, and no death. After the inoculation of the virus, all individuals in the non-immunized challenge group showed symptoms of infection with the new goose parvovirus NMG21 strain, and their growth and development were delayed. During the 30-day experimental observation period, 4 infected ducks died. The results showed that the immune protection rate of the inactivated vaccine prepared by the present invention can reach 100%, with significant effects.

[0089] In summary, the novel goose parvovirus NMG21 strain inactivated vaccine prepared by the present invention is an inactivated vaccine with relatively ideal immune effect, which can effectively prevent the occurrence of the novel goose parvovirus characterized by "neurological symptoms, feather loss, short beak and long tongue".

[0090] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification made within the spirit and principle of the present application shall be deemed to be appropriate.

[0091] Equivalent replacements, improvements, etc. should all be included in the protection scope of this application.

Claims

1. A novel goose parvovirus, characterized in that: The new goose parvovirus was deposited in the General Microbiology Center of China Culture Collection Administration on October 24, 2024, with the deposit number: CGMCC No.46205.

2. The novel goose parvovirus according to claim 1, characterized in that: The complete genome nucleotide sequence of the novel goose parvovirus is shown in SEQ ID NO:

1.

3. Use of the novel goose parvovirus according to claim 1 in preparing a vaccine for preventing the novel goose parvovirus.

4. The use according to claim 3, characterized in that: The vaccine is any one of an inactivated vaccine, a live attenuated vaccine, a subunit vaccine or a genetically engineered vaccine.

5. A novel inactivated goose parvovirus vaccine, characterized in that: The novel goose parvovirus inactivated vaccine comprises the inactivated novel goose parvovirus according to claim 1.

6. The novel goose parvovirus inactivated vaccine according to claim 5, characterized in that: The novel inactivated goose parvovirus vaccine further comprises a pharmaceutically acceptable adjuvant.

7. The novel goose parvovirus inactivated vaccine according to claim 5, characterized in that: The preparation method of the novel goose parvovirus inactivated vaccine comprises the following steps: (1) inoculating the novel goose parvovirus according to claim 1 into DEF cells in the S phase to obtain a virus solution, and after inactivation, obtaining an inactivated antigen solution; (2) adding Tween-80 to the inactivated antigen solution, oscillating and mixing until Tween-80 is completely dissolved to obtain an aqueous phase, and then mixing white oil, aluminum stearate and Span-80 evenly and sterilizing to obtain an oil phase; (3) The oil phase and the water phase were emulsified in a volume ratio of 2:1 to prepare a new goose parvovirus inactivated vaccine.

8. The novel goose parvovirus inactivated vaccine according to claim 7, characterized in that: TCID of the novel goose parvovirus in the virus solution in step (1) 50 For 10 -7.39 / 0.2mL.

9. The novel goose parvovirus inactivated vaccine according to claim 7, characterized in that: The volume ratio of the inactivated antigen solution to the added Tween-80 in step (2) is (24-49):1.

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