KR20220043366A - Vaccine composition for dog canine parvovirus using canine parvovirus type 2 and there uses thereof - Google Patents

Abstract

The present invention relates to a vaccine composition for mammalian parvovirus comprising canine parvovirus type 2 isolated in South Korea or a subculture thereof and a method for preventing parvovirus infection using the same. In addition, the present invention relates to a method for preparing a vaccine composition for mammalian parvovirus, which comprises a step of isolating parvovirus from a sample obtained from the nasal cavity of a mammal and attenuating or inactivating the same. The present invention provides a live or dead vaccine composition which is obtained by isolating canine parvovirus type 2b in South Korea, attenuating or inactivating the same, so as to prevent parvovirus infection more effectively than the CPV-2 vaccine strain currently used in South Korea while having low cytotoxicity. In addition, the present invention can be used as a vaccine for animals other than companion dogs which may be infected with CPV-2 or variants thereof.

Description

Composition for canine parvovirus vaccine using canine parvovirus type 2 and uses thereof {Vaccine composition for dog canine parvovirus using canine parvovirus type 2 and there uses thereof}

The present invention relates to a composition for a mammalian parvovirus vaccine comprising canine parvovirus type 2b or a subculture thereof isolated in Korea and a method for preventing parvovirus infection using the same.

The present invention also relates to a method for preparing a composition for a mammalian parvovirus vaccine, comprising the step of isolating a parvovirus from a sample obtained from the nasal cavity of a mammal and attenuating or inactivating it.

Canine parvovirus (CPV) is an enteric pathogen that infects mammals, particularly dogs. Parvovirus can cause acute diarrhea, fever and leukopenia in puppies older than 4 to 5 weeks of age and myocardial disease in younger puppies. It started to occur in the United States, Belgium, or Australia in the late 1970s and is now spreading worldwide. In Korea, it first appeared in Gyeonggi-do in the 1980s, and according to a recent study, it was found that parvovirus was the cause of about 89% of dogs with enteritis symptoms in Korea.

Canine parvovirus-2 (CPV-2) belongs to the Parvoviridae family and has a single-stranded DNA gene, so it is a virus with very high mutability. There are two types of capsid proteins of parvovirus, VP1 and VP2, of which VP2 is the main immunogenic capsid protein of parvovirus. Currently, CPV isolated in Korea is mostly identified as CPV-2a and CPV-2b, which are mutants derived from CPV-2.

Korean Patent Laid-Open Publication No. 10-2005-0011909 discloses that parvovirus is transformed using a vector so that the antibody binds to the parvovirus antigen better, thereby effectively preventing or treating parvovirus infection, neutralizing canine parvovirus. To a VP2 recombinant protein comprising an antigenic determinant and a vaccine composition comprising the same. However, there have been no reports of studies that attempt to attenuate or inactivate a parvovirus isolated in Korea and use it as a vaccine against CPV-2 or its mutant CPV-2b.

It has been more than 40 years since the CPV-2 vaccine strain currently in use in Korea was developed, and was developed based on CPV-2 before it was derived into another mutant strain. Types CPV-2a and CPV-2b derived from CPV-2 do not properly form neutralizing antibodies from conventional CPV-2, whereas CPV-2a and CPV-2b produce neutralizing antibodies between CPV-2 and mutant strains. it is known If the vaccine is not updated according to the mutant, even if the vaccine is vaccinated, the mutant is not effective, so parvovirus infection cannot be prevented. That is, the CPV-2 vaccine strain currently in use in Korea has a problem in that it does not exhibit a sufficient effect on the CPV-2 mutant strain.

Accordingly, as a result of the present inventors' efforts to provide an effective vaccine for the prevention of CPV-2 mutant that is prevalent in Korea, when CPV-2b isolated directly in Korea is sufficiently attenuated and inactivated, its toxicity is weakened and canine parvovirus As a vaccine against , it was confirmed that it is more effective than the vaccine strain currently used in Korea and completed the present invention.

In order to achieve the above object, the present invention provides a canine parvovirus type 2b (CPV-2b) or passage thereof comprising a VP-2 (viral capsid peptide-2) protein consisting of the amino acid sequence of SEQ ID NO: 3 It is possible to provide a composition for a mammalian parvovirus vaccine comprising a culture.

According to a preferred embodiment of the present invention, the canine parvovirus type 2b may be a live cell or a dead cell.

According to a preferred embodiment of the present invention, the VP-2 protein may be encoded by the nucleotide sequence of SEQ ID NO: 4.

According to a preferred embodiment of the present invention, the subculture may be canine parvovirus type 2b obtained by subculture of canine parvovirus type 2b 90 to 150 times.

According to a preferred embodiment of the present invention, the subculture is canine parvovirus type 2b comprising a VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 5; or canine parvovirus type 2b of accession number KCTC18847P; may be

According to a preferred embodiment of the present invention, the VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 5 may be encoded by the nucleotide sequence of SEQ ID NO: 6.

According to a preferred embodiment of the present invention, the mammal may be any one or more selected from the group consisting of dogs, cats, foxes, wolves, pigs and skunks.

In addition, the present invention provides a method for preventing parvovirus infection by administering the composition for a vaccine to a mammal other than a human.

In addition, the present invention comprises the steps of i) obtaining a sample from the nasal cavity of a mammal;

ii) isolating a parvovirus comprising the VP-2 protein of SEQ ID NO: 3 from the sample of step i); and

iii) subculturing the parvovirus of step ii) 90 to 150 times;

It provides a method for preparing a composition for a mammalian parvovirus vaccine comprising a.

According to a preferred embodiment of the present invention, iv) inactivating the parvovirus by treating the subculture of step iii) with formaldehyde and glycine at a molar concentration ratio of 3:1 to 1:4;

It may be to include additionally.

According to a preferred embodiment of the present invention, the mammal of step i) may be any one or more selected from the group consisting of dogs, cats, foxes, wolves, pigs and skunks.

The present invention provides a composition for live or dead poison vaccine by attenuating or inactivating canine parvovirus type 2b isolated in Korea to prevent parvovirus infection more effectively than the CPV-2 vaccine currently used in Korea while having low cytotoxicity. make it possible

In addition, the present invention can be used as a vaccine for animals other than dogs that can be infected with CPV-2 or a mutant thereof.

1 shows CRFK cells not infected with canine parvovirus type 2b (A) and CRFK cells infected with canine parvovirus type 2b (B).

Hereinafter, the terms of the present invention will be described.

The "viral capsid peptide-2 (VP-2) protein" of the present invention is a parvovirus capsid protein (VP) and is a major immunogenic parvovirus capsid protein. Two main types are known, VP-1 and VP-2.

The "live vaccine" of the present invention is also called an attenuated vaccine, and refers to a vaccine using a virus or bacteria that has the ability to self-reproduce and induce immunity by modifying a part of a virus or bacteria that causes a disease, but has the ability to cause toxicity . The modification may be performed by UV irradiation, chemical treatment, cell culture, egg inoculation, or inoculation with live animals.

The "dead poison vaccine" of the present invention refers to a vaccine containing mainly inactivated bacteria using chemicals. Because the fungus is not alive, it cannot reproduce and cannot cause infection. In the case of sadox vaccine, it is usually administered several times. The first administration does not cause a protective immune response, and the immune response occurs from the second administration or more.

Hereinafter, the present invention will be described in more detail.

As described above, the CPV-2 vaccine strain used as a vaccine against canine parvovirus type 2b (CPV-2b) in Korea has been developed for a considerable period of time, and CPV-2b, a mutant strain of CPV-2 that is currently prevalent in Korea. There was a limitation that the quarantine effect was not sufficient. In addition, studies on the attenuation or inactivation of a CPV-2b isolate newly isolated in Korea other than the previously known CPV-2b sequence to provide a vaccine composition were only insignificant.

When the canine parvovirus type 2b (CPV-2b) isolate isolated in Korea according to the present invention is attenuated or inactivated and used for the manufacture of a vaccine for parvovirus prevention, CPV-2 or CPV-2b is weak in cytotoxicity. It is effective as a composition for mammalian parvovirus vaccine because it can provide an excellent prevention effect against

Accordingly, the present invention includes a canine parvovirus type 2b (CPV-2b) comprising a VP-2 (viral capsid peptide-2) protein consisting of the amino acid sequence of SEQ ID NO: 3 or a subculture thereof. A composition for a mammalian parvovirus vaccine is provided.

CPV-2b is a mutant of CPV-2, a parvovirus that mainly infects dogs, and is generally known to be mutated from feline panleukopeni virus (FPV). In addition to dogs, it can be infected with mammals such as carnivores, but it is known that it does not infect humans.

According to a preferred embodiment of the present invention, the canine parvovirus type 2b may be a live cell or a dead cell. When included as live cells, it can be used as a composition for live poison vaccine, and when included as dead cells, it can be used as a composition for dead poison vaccine.

In order to be included as live cells, if used as a vaccine composition directly without an attenuation process, the pathogenic or cytotoxicity of the bacteria is strong and can cause disease beyond the enhancement of immunity, so a step of attenuating live cells is required. In the case of attenuation through cell culture, if cell degeneration is confirmed by bacteria, it is considered that the cytotoxicity of the bacteria is not sufficiently attenuated, so cells are destroyed through freeze-thaw, and the infected bacteria are released and cell culture is performed again. In the present invention, the parvovirus was attenuated using a subculture method in which cells were cultured 90 times or more.

In order to include the bacteria as dead cells, a step of inactivating the bacteria using a chemical substance or heat is required. In the present invention, formaldehyde and glycine were used together to inactivate parvovirus.

According to a preferred embodiment of the present invention, the VP-2 protein may be encoded by the nucleotide sequence of SEQ ID NO: 4. Canine parvovirus type 2b, encoded by the nucleotide sequence of SEQ ID NO: 4 and containing the VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 3, was isolated from nasal secretions obtained from the nasal passages of dogs distributed in Korea. In the case of isolating the parvovirus from the sample obtained from the nasal passages of dogs, it is easier than in the case of isolating the parvovirus from the feces of dogs.

According to a preferred embodiment of the present invention, the subculture may be canine parvovirus type 2b obtained by subculture of canine parvovirus type 2b 90 to 150 times. More preferably, the subculture may be canine parvovirus type 2b obtained by subculture of canine parvovirus type 2b 95 to 130 times, and more preferably, canine parvovirus type 2b obtained by passage 98 or 120 times. It may be virus type 2b. The canine parvovirus type 2b obtained by passage 98 contains the VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 5, and the canine parvovirus type 2b obtained by passage 120 times is a biological resource of the Korea Institute of Biotechnology and Biotechnology. It was deposited with the Center on September 16, 2020 (Accession No. KCTC18847P). The VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 5 may be encoded by the nucleotide sequence of SEQ ID NO: 6.

The canine parvovirus type 2b of the accession number KCTC18847P is preferably obtained by subculturing the canine parvovirus type 2b containing the VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 3 110 to 130 times, most preferably It is preferable to pass 120 times.

According to a preferred embodiment of the present invention, the mammal may be any one or more selected from the group consisting of dogs, cats, foxes, wolves, pigs and skunks, preferably dogs.

According to a preferred embodiment of the present invention, the vaccine may be a live attenuated vaccine, a dead poison vaccine, a subunit vaccine, a synthetic vaccine, or a genetically engineered vaccine, but it is preferably a live attenuated vaccine or a dead poison vaccine.

The vaccine composition may further include one or more selected from the group consisting of a solvent, an adjuvant, and an excipient. The solvent includes physiological saline or distilled water, the immune adjuvant includes Freund's incomplete or complete adjuvant, aluminum hydroxide gel, and vegetable and mineral oils, and the excipient includes aluminum phosphate and aluminum hydroxide. side or aluminum potassium sulfate (alum). In addition, the vaccine composition may further include a stabilizer, an inactivating agent, an antibiotic or a preservative, and may be used by mixing with distilled water, a buffer solution, etc. depending on the route of administration of the vaccine. However, the present invention is not limited thereto, and may include all known substances used in the preparation of vaccines well known to those of ordinary skill in the art.

The vaccine composition may be administered in a pharmaceutically effective amount. A pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment. Sensitivity to, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined. In addition, it may be administered as an individual therapeutic agent or may be administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. and may be administered single or multiple. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

In addition, the present invention provides a method for preventing parvovirus infection by administering the composition for a vaccine to a mammal other than a human.

The administration may be administered through an administration route such as oral, intramuscular, or subcutaneous, but is not limited thereto.

The prevention refers to any action that suppresses or delays the onset of parvovirus infection due to the administration of the vaccine composition.

In addition, the present invention comprises the steps of i) obtaining a sample from the nasal cavity of a mammal;

ii) isolating a parvovirus comprising the VP-2 protein of SEQ ID NO: 3 from the sample of step i); and

iii) subculturing the parvovirus of step ii) 90 to 150 times;

It provides a method for preparing a composition for a mammalian parvovirus vaccine comprising a.

According to a preferred embodiment of the present invention, the preparation method comprises the steps of iv) treating the subculture of step iii) with formaldehyde and glycine at a molar concentration ratio of 3:1 to 1:4 to inactivate parvovirus ; It may be to include additionally. More preferably, it is preferable to treat formaldehyde and glycine in a molar ratio of 1:1 to 1:4, and even more preferably, it is most preferable to treat formaldehyde and glycine in a molar ratio of 3:4.

The mammal of step i) of the present invention is preferably any one or more selected from the group consisting of dogs, cats, foxes, wolves, pigs and skunks, but is not limited in the case of mammals other than humans such as carnivores this can be

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.

Isolation and identification of canine parvovirus

The virus was isolated from a nasal swab sample that is distributed in the domestic market and obtained from the nasal cavity of a dog suspected of being infected with parvovirus, and it was attempted to confirm that it was parvovirus type 2b (CPV-2b).

Specifically, the dog's nasal secretions were dissolved in phosphate buffered saline (PBS), and then filtered through a 0.2 μm filter to isolate parvovirus. The isolated parvovirus DNA was extracted and compared with the sequence of canine parvovirus capsid protein 2 (VP2) through polymerase chain reaction (PCR).

As a result, it was confirmed that the virus obtained from the nasal secretions of dogs was a parvovirus of CPV-2b that is prevalent in Korea.

Passage of canine parvovirus

In order to attenuate the CPV-2b obtained in [Example 1], it was attempted to subculture the infected cells.

Specifically, crandell rees feline kidney (CRFK) cells were subcultured using MEM (Minimum Essential medium) medium containing 10% Fetal Bovine Serum (FBS). After removing the medium when the CRFK cells cover about 80% of the flask area, 0.1 MOI (Multiplicity of infection). After incubation at 37 ° C. for 1 hour, the virus was removed and placed in MEM medium for maintenance. When the cytopathic effect of CRFK cells due to infection with canine parvovirus was confirmed 2-3 days after infection, the freeze-thaw process was repeated three times, and then the virus was harvested. This was repeated 120 times, that is, 120 passages. Among them, the sequences of parvoviruses that were passaged 98 and passage 120 were confirmed. The following [Table 1] shows the previously known VP-2 (CPV-LZ2) sequence of CPV-2b, the VP-2 sequence of the canine parvovirus obtained in [Example 1] and the case of 98 passages thereof. The VP-2 sequence is shown.

Parvovirus sequence types SEQ ID NO: CPV-LZ2 VP-2 amino acid sequence One CPV-LZ2 VP-2 nucleotide sequence 2 CPV-2b amino acid sequence of [Example 1] 3 CPV-2b nucleotide sequence of [Example 1] 4 Amino acid sequence in which CPV-2b of [Example 1] was passaged 98 times 5 [Example 1] CPV-2b nucleotide sequence passaged 98 times 6

As a result, compared with the previously known CPV-2b sequence, CPV-2b obtained in [Example 1] had a total of 4 amino acid sequences and a total of 15 nucleotide sequences. When the CPV-2b obtained in [Example 1] was subcultured 98 times, a total of 12 amino acid sequences and a total of 23 nucleotide sequences were different. That is, as the number of subcultures increased, it could be confirmed that the difference from the known sequence increased, and it could be expected that parvovirus attenuation occurred.

Confirmation of Cytotoxicity of Attenuated Canine Parvovirus

An attempt was made to determine whether attenuation was sufficiently performed on the CPV-2b parvovirus subcultured in [Example 2].

Specifically, the cytotoxicity of the CPV-2b parvovirus subcultured in [Example 2] was experimentally confirmed.

As a result, it was confirmed that the cytotoxicity of the parvovirus was sufficiently weakened in both the CPV-2b parvoviruses that were cultured at passage 98 or 120 times.

Inactivation of canine parvovirus

The CPV-2b parvovirus subcultured in [Example 2] was inactivated, and the presence or absence of cytotoxicity was checked.

Specifically, the CPV-2b virus passaged 98 or 120 times in [Example 2] was mixed with 0.03 M formaldehyde and 0.04 M glycine, and reacted at 37° C. for 24 hours without light. . After that, it was checked whether cytotoxicity appeared.

As a result, it was confirmed that neither the CPV-2b parvovirus treated with formaldehyde and glycine with passage 98 or passage 120 nor CPV-2b parvovirus exhibited cytotoxicity.

[Entrusted Organization]

Name of depositary institution: Korea Research Institute of Bioscience and Biotechnology

Accession number: KCTC18847P

Deposit date: 20200916

SEQUENCE LISTING <110> KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP <120> Vaccine composition for dog canine parvovirus using canine parvovirus type 2 and there uses thereof <130> 1063975 <160> 6 <170> PatentIn version 3.2 <210> 1 <211> 727 <212> PRT <213> Artificial <220> <223> CPV-LZ2_VP-2_amino acid <400> 1 Met Ala Pro Pro Ala Lys Arg Ala Arg Arg Gly Leu Val Pro Pro Gly 1 5 10 15 Tyr Lys Tyr Leu Gly Pro Gly Asn Ser Leu Asp Gln Gly Glu Pro Thr 20 25 30 Asn Pro Ser Asp Ala Ala Ala Lys Glu His Asp Glu Ala Tyr Ala Ala 35 40 45 Tyr Leu Arg Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Ser Pro Ala Asp 50 55 60 Gln Arg Phe Ile Asp Gln Thr Lys Asp Ala Lys Asp Trp Gly Gly Lys 65 70 75 80 Ile Gly His Tyr Phe Phe Arg Ala Lys Lys Ala Ile Ala Pro Val Leu 85 90 95 Thr Asp Thr Pro Asp His Pro Ser Thr Ser Arg Pro Thr Lys Pro Thr 100 105 110 Lys Arg Ser Arg Pro Pro His Ile Phe Ile Asn Leu Ala Lys Lys 115 120 125 Lys Lys Ala Gly Ala Gly Gln Val Lys Arg Asp Asn Leu Ala Pro Met 130 135 140 Ser Asp Gly Ala Val Gln Pro Asp Gly Gly Gln Pro Ala Val Arg Asn 145 150 155 160 Glu Arg Ala Thr Gly Ser Gly Asn Gly Ser Gly Gly Gly Gly Gly Gly 165 170 175 Gly Ser Gly Gly Val Gly Ile Ser Thr Gly Thr Phe Asn Asn Gln Thr 180 185 190 Glu Phe Lys Phe Leu Glu Asn Gly Trp Val Glu Ile Thr Ala Asn Ser 195 200 205 Ser Arg Leu Val His Leu Asn Met Pro Glu Ser Glu Asn Tyr Arg Arg 210 215 220 Val Val Val Asn Asn Leu Asp Lys Thr Ala Val Asn Gly Asn Met Ala 225 230 235 240 Leu Asp Asp Thr His Ala Gln Ile Val Thr Pro Trp Ser Leu Val Asp 245 250 255 Ala Asn Ala Trp Gly Val Trp Phe Asn Pro Gly Asp Trp Gln Leu Ile 260 265 270 Val Asn Thr Met Ser Glu Leu His Leu Val Ser Phe Glu Gln Glu Ile 275 280 285 Phe Asn Val Val Leu Lys Thr Val Ser Glu Ser Ala Thr Gln Pro Pro 290 295 300 Thr Lys Val Tyr Asn Asn Asp Leu Thr Ala Ser Leu Met Val Ala Leu 305 310 315 320 Asp Ser Asn Asn Thr Met Pro Phe Thr Pro Ala Ala Met Arg Ser Glu 325 330 335 Thr Leu Gly Phe Tyr Pro Trp Lys Pro Thr Ile Pro Thr Pro Trp Arg 340 345 350 Tyr Tyr Phe Gln Trp Asp Arg Thr Leu Ile Pro Ser His Thr Gly Thr 355 360 365 Ser Gly Thr Pro Thr Asn Ile Tyr His Gly Thr Asp Pro Asp Asp Val 370 375 380 Gln Phe Tyr Thr Ile Glu Asn Ser Val Pro Val His Leu Leu Arg Thr 385 390 395 400 Gly Asp Glu Phe Ala Thr Gly Thr Phe Tyr Phe Asp Cys Lys Pro Cys 405 410 415 Arg Leu Thr His Thr Trp Gln Thr Asn Arg Ala Leu Gly Leu Pro Pro 420 425 430 Phe Leu Asn Ser Leu Pro Gln Ala Glu Gly Gly Thr Asn Phe Gly Tyr 435 440 445 Ile Gly Val Gln Gln Asp Lys Arg Arg Gly Val Thr Gln Met Gly Asn 450 455 460 Thr Asn Ile Ile Thr Glu Ala Thr Ile Met Arg Pro Ala Glu Val Gly 465 470 475 480 Tyr Ser Ala Pro Tyr Tyr Ser Phe Glu Ala Ser Thr Gln Gly Pro Phe 485 490 495 Lys Thr Pro Ile Ala Ala Gly Arg Gly Gly Ala Gln Thr Asp Glu Asn 500 505 510 Gln Ala Ala Asp Gly Asp Pro Arg Tyr Ala Phe Gly Arg Gln His Gly 515 520 525 Gln Lys Thr Thr Thr Thr Gly Glu Thr Pro Glu Arg Phe Thr Tyr Ile 530 535 540 Ala His Gln Asp Thr Gly Arg Tyr Pro Glu Gly Asp Trp Ile Gln Asn 545 550 555 560 Ile Asn Phe Asn Leu Pro Val Thr Asp Asp Asn Val Leu Leu Pro Thr 565 570 575 Asp Pro Ile Gly Gly Lys Ala Gly Ile Asn Tyr Thr Asn Ile Phe Asn 580 585 590 Thr Tyr Gly Pro Leu Thr Ala Leu Asn Asn Val Pro Pro Val Tyr Pro 595 600 605 Asn Gly Gln Ile Trp Asp Lys Glu Phe Asp Thr Asp Leu Lys Pro Arg 610 615 620 Leu His Val Asn Ala Pro Phe Val Cys Gln Asn Asn Cys Pro Gly Gln 625 630 635 640 Leu Phe Val Lys Val Ala Pro Asn Leu Thr Asn Glu Tyr Asp Pro Asp 645 650 655 Ala Ser Ala Asn Met Ser Arg Ile Val Thr Tyr Ser Asp Phe Trp Trp 660 665 670 Lys Gly Lys Leu Val Phe Lys Ala Lys Leu Arg Ala Ser His Thr Trp 675 680 685 Asn Pro Ile Gln Gln Met Ser Ile Asn Val Asp Asn Gln Phe Asn Tyr 690 695 700 Val Pro Ser Asn Ile Gly Gly Met Lys Ile Val Tyr Glu Lys Ser Gln 705 710 715 720 Leu Ala Pro Arg Lys Leu Tyr 725 <210> 2 <211> 2184 <212> DNA <213> Artificial <220> <223 > CPV-LZ2_VP-2_nucleotide <400> 2 atggcacctc cggcaaagag agccaggaga ggacttgtgc ctccaggtta taaatatctt 60 gggcctggga acagtcttga ccaaggagaa ccaactaacc cttctgacgc cgctgcaaaa 120 gaacacgacg aagcttacgc tgcttatctt cgctctggta aaaacccata cttatatttc 180 tcgccagcag atcaacgctt tatagatcaa actaaggacg ctaaagattg gggggggaaa 240 ataggacatt atttttttag agctaaaaag gcaattgctc cagtattaac tgatacacca 300 gatcatccat caacatcaag accaacaaaa ccaactaaaa gaagtagacc accacctcat 360 attttcatca atcttgcaaa aaaaaaaaaa gccggtgcag gacaagtaaa aagagacaat 420 cttgcaccaa tgagtgatgg agcagttcaa ccagacggtg gtcagcctgc tgtcagaaat 480 gaaagagcta caggatctgg gaacgggtct ggaggcgggg gtggtggtgg ttctgggggt 540 gtggggattt ctacgggtac tttcaataat cagacggaat ttaaattttt ggaaaacgga 600 tgggtggaaa tcacagcaaa ctcaagcaga cttgt acatt taaatatgcc agaaagtgaa 660 aattatagaa gagtggttgt aaataatttg gataaaactg cagttaacgg aaacatggct 720 ttagatgata cccatgcaca aattgtaaca ccttggtcat tggttgatgc aaatgcttgg 780 ggagtttggt ttaatccagg agattggcaa ctaattgtta atactatgag tgagttgcat 840 ttagttagtt ttgaacaaga aatttttaat gttgttttaa agactgtttc agaatctgct 900 actcagccac caactaaagt ttataataat gatttaactg catcattgat ggttgcatta 960 gatagtaata atactatgcc atttactcca gcagctatga gatctgagac attgggtttt 1020 tatccatgga aaccaaccat accaactcca tggagatatt attttcaatg ggatagaaca 1080 ttaataccat ctcatactgg aactagtggc acaccaacaa atatatacca tggtacagat 1140 ccagatgatg ttcaatttta cactattgaa aattctgtgc cagtacactt actaagaaca 1200 ggtgatgaat ttgctacagg aacattttat tttgattgta aaccatgtag actaacacac 1260 acatggcaaa caaatagagc attgggctta ccaccatttc taaattcttt gcctcaagct 1320 gaaggaggta ctaactttgg ttatatagga gttcaacaag ataaaagacg tggtgtaact 1380 caaatgggaa atacaaacat tattactgaa gctactatca tgagaccagc tgaggttggt 1440 tatagtgcac catattattc ttttgaggcg tctacacaag ggccat ttaa aacacctatt 1500 gcagcaggac gggggggagc gcaaacagat gaaaatcaag cagcagatgg tgatccaaga 1560 tatgcatttg gtagacaaca tggtcaaaaa actaccacaa caggagaaac acctgagaga 1620 tttacatata tagcacatca agatacagga agatatccag aaggagattg gattcaaaat 1680 attaacttta accttcctgt aacagatgat aatgtattgc taccaacaga tccaattgga 1740 ggtaaagcag gaattaacta tactaatata tttaatactt atggtccttt aactgcatta 1800 aataatgtac caccagttta tccaaatggt caaatttggg ataaagaatt tgatactgac 1860 ttaaaaccaa gacttcatgt aaatgcacca tttgtttgtc aaaataattg tcctggtcaa 1920 ttatttgtaa aagttgcgcc taatttaaca aatgaatatg atcctgatgc atctgctaat 1980 atgtcaagaa ttgtaactta ctcagatttt tggtggaaag gtaaattagt atttaaagct 2040 aaactaagag cctctcatac ttggaatcca attcaacaaa tgagtattaa tgtagataac 2100 caatttaact atgtaccaag taatattgga ggtatgaaaa ttgtatatga aaaatctcaa 2160 ctagcaccta gaaaattata ctaa 2184 <210> 3 <211> 727 <212> PRT <213> Artificial <220> <223> CPV-2b_embodiment 1_amino acid <400> 3 Met Ala Pro Pro Ala Lys Arg Ala Arg Arg Gly Leu Val Pro Pro Gly 1 5 10 15 Tyr Lys Tyr Leu Gly Pro Gly Asn Ser Leu Asp Gln Gly Glu Pro Thr 20 25 30 Asn Pro Ser Asp Ala Ala Ala Lys Glu His Asp Glu Ala Tyr Ala Ala 35 40 45 Tyr Leu Arg Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Ser Pro Ala Asp 50 55 60 Gln Arg Phe Ile Asp Gln Thr Lys Asp Ala Lys Asp Trp Gly Gly Lys 65 70 75 80 Ile Gly His Tyr Phe Phe Arg Ala Lys Lys Ala Ile Ala Pro Val Leu 85 90 95 Thr Asp Thr Pro Asp His Pro Ser Thr Ser Arg Pro Thr Lys Pro Thr 100 105 110 Lys Arg Ser Lys Pro Pro Pro His Ile Phe Ile Asn Leu Ala Lys Lys 115 120 125 Lys Lys Ala Gly Ala Gly Gln Val Lys Arg Asp Asn Leu Ala Pro Met 130 135 140 Ser Asp Gly Ala Val Gln Pro Asp Gly Gly Gln Pro Ala Val Arg Asn 145 150 155 160 Glu Arg Ala Thr Gly Ser Gly Asn Gly Ser Gly Gly Gly Gly Gly Gly Gly 165 170 175 Gly Ser Gly Gly Val Gly Ile Ser Thr Gly Thr Phe Asn As n Gln Thr 180 185 190 Glu Phe Lys Phe Leu Glu Asn Gly Trp Val Glu Ile Thr Ala Asn Ser 195 200 205 Ser Arg Leu Val His Leu Asn Met Pro Glu Ser Glu Asn Tyr Arg Arg Arg 210 215 220 Val Val Val Asn Asn Leu Asp Lys Thr Ala Val Asn Gly Asn Met Ala 225 230 235 240 Leu Asp Asp Thr His Ala Gln Ile Val Thr Pro Trp Ser Leu Val Asp 245 250 255 Ala Asn Ala Trp Gly Val Trp Phe Asn Pro Gly Asp Trp Gln Leu Ile 260 265 270 Val Asn Thr Met Ser Glu Leu His Leu Val Ser Phe Glu Gln Glu Ile 275 280 285 Phe Asn Val Val Leu Lys Thr Val Ser Glu Ser Ala Thr Gln Pro 290 295 300 Thr Lys Val Tyr Asn Asn Asp Leu Thr Ala Ser Leu Met Val Ala Leu 305 310 315 320 Asp Ser Asn Asn Thr Met Pro Phe Thr Pro Al a Ala Met Arg Ser Glu 325 330 335 Thr Leu Gly Phe Tyr Pro Trp Lys Pro Thr Ile Pro Thr Pro Trp Arg 340 345 350 Tyr Tyr Phe Gln Trp Asp Arg Thr Leu Ile Pro Ser His Thr Gly Thr 355 360 365 Ser Gly Thr Pro Thr Asn Ile Tyr His Gly Thr Asp Pro Asp Asp Val 370 375 380 Gln Phe Tyr Thr Ile Glu Asn Ser Val Pro Val His Leu Leu Arg Thr 385 390 395 400 Gly Asp Glu Phe Ala Thr Gly Thr Phe Phe Phe Asp Cys Lys Pro Cys 405 410 415 Arg Leu Thr His Thr Trp Gln Thr Asn Arg Ala Leu Gly Leu Pro Pro 420 425 430 Phe Leu Asn Ser Leu Pro Gln Ala Glu Gly Gly Thr Asn Phe Gly Tyr 435 440 445 Ile Gly Val Gln Gln Asp Lys Arg Arg Gly Val Thr Gln Met Gly Asn 450 455 460 Thr Asn Tyr Ile Thr Glu Ala Thr Ile Met Arg Pro Al a Glu Val Gly 465 470 475 480 Tyr Ser Ala Pro Tyr Tyr Ser Phe Glu Ala Ser Thr Gln Gly Pro Phe 485 490 495 Lys Thr Pro Ile Ala Ala Gly Arg Gly Gly Ala Gln Thr Asp Glu Asn 500 505 510 Gln Ala Ala Asp Gly Asp Pro Arg Tyr Ala Phe Gly Arg Gln His Gly 515 520 525 Gln Lys Thr Thr Thr Thr Gly Glu Thr Pro Glu Arg Phe Thr Tyr Ile 530 535 540 Ala His Gln Asp Thr Gly Arg Tyr Pro Glu Gly Asp Trp Ile Gln Asn 545 550 555 560 Ile Asn Phe Asn Leu Pro Val Thr Asp Asp Asn Val Leu Leu Pro Thr 565 570 575 Asp Pro Ile Gly Gly Lys Thr Gly Ile Asn Tyr Thr Asn Ile Phe Asn 580 585 590 Thr Tyr Gly Pro Leu Thr Ala Leu Asn Asn Val Pro Val Tyr Pro 595 600 605 Asn Gly Gln Ile Trp Asp Lys Glu Phe As p Thr Asp Leu Lys Pro Arg 610 615 620 Leu His Val Asn Ala Pro Phe Val Cys Gln Asn Asn Cys Pro Gly Gln 625 630 635 640 Leu Phe Val Lys Val Ala Pro Asn Leu Thr Asn Glu Tyr Asp Pro Asp 645 650 655 Ala Ser Ala Asn Met Ser Arg Ile Val Thr Tyr Ser Asp Phe Trp Trp 660 665 670 Lys Gly Lys Leu Val Phe Lys Ala Lys Leu Arg Ala Ser His Thr Trp 675 680 685 Asn Pro Ile Gln Gln Met Ser Ile Asn Val Asp Asn Gln Phe Asn Tyr 690 695 700 Val Pro Ser Asn Ile Gly Gly Met Lys Ile Val Tyr Glu Lys Ser Gln 705 710 715 720 Leu Ala Pro Arg Lys Leu Tyr 725 <210> 4 <211> 2184 <212> DNA <213> Artificial <220> <223> CPV-2b_embodiment 1_nucleotide <400> 4 atggcacctc cggcaaagag agccaggaga ggacttgtgc ctccaggtta taaatatctt 60 gggcctggga acagtcttga ccaaggagaa ccaactaacc cttctgacgc cgctgcaaaa 120 gaacacgacg aagcttacgc tgcttatctt cgctctggta aaaacccata cttatatttc 180 tcgccagcag atcaacgctt tatagatcaa actaaggacg ctaaagattg gggggggaaa 240 ataggacatt atttttttag agctaaaaag gcaattgctc cagtattaac tgatacacca 300 gatcatccat caacatcaag accaacaaaa ccaactaaaa gaagtaaacc accacctcat 360 attttcatca atcttgcaaa aaaaaaaaaa gccggtgcag gacaagtaaa aagagacaat 420 cttgcaccaa tgagtgatgg agcagttcaa ccagacggtg gtcagcctgc tgtcagaaat 480 gaaagagcta caggatctgg gaacgggtct ggaggcgggg gtggtggtgg ttctgggggt 540 gtggggattt ctacgggtac tttcaataat cagacggaat ttaaattttt ggaaaacgga 600 tgggtggaaa tcacagcaaa ctcaagcaga cttgtacatt taaatatgcc agaaagtgaa 660 aattatagaa gagtggttgt aaataatttg gataagactg cagttaacgg aaacatggct 720 ttagatgata ctcatgcaca aattgtaaca ccttggtcat tggttgatgc aaatgcttgg 780 ggagtttggt ttaatccagg agattggcaa ctaattgtta atactatgag tgagttgcat 840 ttagttagtt ttgaacaaga aatttttaat gttgttttaa agactgtttc agaatctgct 900 actcagccac caactaaagt ttataataat gatttaactg catcattgat ggttgcatt a 960 gatagtaata atactatgcc atttactcca gcagctatga gatctgagac attgggtttt 1020 tatccatgga aaccaaccat accaactcca tggagatatt attttcaatg ggatagaaca 1080 ttaataccat ctcatactgg aactagtggc acaccaacaa atatatacca tggtacagat 1140 ccagatgatg ttcaatttta tactattgaa aattctgtgc cagtacactt actaagaaca 1200 ggtgatgaat ttgctacagg aacatttttt tttgattgta aaccatgtag actaacacat 1260 acatggcaaa caaatagagc attgggctta ccaccatttc taaattcttt gcctcaagct 1320 gaaggaggta ctaactttgg ttatatagga gttcaacaag ataaaagacg tggtgtaact 1380 caaatgggaa atacaaacta tatcactgaa gctactatta tgagaccagc tgaggttggt 1440 tatagtgcac catattattc ttttgaggcg tctacacaag ggccgtttaa aacacctatt 1500 gcagcaggac gggggggagc gcaaacagat gaaaatcaag cagcagatgg tgatccaaga 1560 tatgcatttg gtagacaaca tggtcaaaaa actaccacaa caggagaaac acctgagaga 1620 tttacatata tagcacatca agatacagga agatatccag aaggagattg gattcaaaat 1680 attaacttta accttcctgt aacagatgat aatgtattgc taccaacaga tccaattgga 1740 ggtaaaacag gaattaacta tactaatata tttaatactt atggtccttt aactgcatta 1800aataatgtac caccagttta tccaaatggt caaatttggg ataaagaatt tgatactgac 1860 ttaaaaccaa gacttcatgt aaatgcacca tttgtttgtc aaaataattg ccctggtcaa 1920 ttatttgtaa aagttgcgcc taatttaaca aatgaatatg atcctgatgc atctgctaat 1980 atgtcaagaa ttgtaactta ctcagatttt tggtggaaag gtaaattagt atttaaagct 2040 aaactaagag cctctcatac ttggaatcca attcaacaaa tgagtattaa tgtagataac 2100 caatttaact atgtaccaag taatattgga ggtatgaaaa ttgtctatga aaaatctcaa 2160 ctagcaccta gaaaattata ttaa 2184 <210> 5 <211> 727 <212> PRT <213> Artificial <220> <223> CPV-2b_embodiment 1_passage 98_amino acid <400> 5 Met Ala Pro Pro Ala Lys Arg Ala Arg Arg Gly Leu Val Pro Pro Gly 1 5 10 15 Tyr Lys Tyr Leu Gly Pro Gly Asn Ser Leu Asp Gln Gly Glu Pro Thr 20 25 30 Asn Pro Ser Asp Ala Ala Ala Lys Glu His Asp Glu Ala Tyr Ala Ala 35 40 45 Tyr Leu Arg Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Ser Pro Ala Asp 50 55 60 Gln Arg Phe Ile Asp Gln Thr Lys Asp Ala Lys Asp Trp Gly Gly Lys 65 70 75 80 Ile Gly His Tyr Phe Phe Arg Ala Lys Lys Ala Ile Ala Pro Val Leu 85 90 95 Thr Asp Thr Pro Asp His Pro Ser Thr Ser Arg Pro Thr Lys Pro Thr 100 105 110 Lys Arg Ser Lys Pro Pro Pro His Ile Phe Ile Asn Leu Ala Lys Lys 115 120 125 Lys Lys Ala Gly Ala Gly Gln Val Lys Arg Asp Asn Leu Ala Pro Met 130 135 140 Ser Asp Gly Ala Val Gln Pro Asp Gly Gly Gln Pro Ala Val Arg Asn 145 150 155 160 Glu Arg Ala Thr Gly Ser Gly Asn Gly Ser Gly Gly Gly Gly Gly Gly 165 170 175 Gly Ser Gly Gly Val Gly Ile Ser Thr Gly Thr Phe Asn Asn Gln Thr 180 185 190 Glu Phe Lys Phe Leu Glu Asn Gly Trp Val Glu Ile Thr Ala Asn Ser 195 200 205 Ser Arg Leu Val His Leu Asn Met Pro Glu Ser Glu Asn Tyr Arg Arg 210 215 220 Val Val Val Asn Asn Leu Asp Lys Thr Ala Val Lys Gly Asn Met Ala 225 230 235 240 Leu Asp Asp Thr His Ala Gln Ile Val Thr Pro Trp Ser Leu Val Asp 245 250 255 Ala Asn Ala Trp Gly Val Trp Phe Asn Pro Gly Asp Trp Gln Leu Ile 260 265 270 Val Asn Thr Met Ser Glu Leu His Leu Val Ser Phe Glu Gln Glu Ile 275 280 285 Phe Asn Val Val Leu Lys Thr Val Ser Glu Ser Ala Thr Gln Pro Pro 290 295 300 Thr Lys Val Tyr Asn Asn Asp Leu Thr Ala Ser Leu Met Val Ala Leu 305 310 315 320 Asp Ser Asn Asn Thr Met Pro Phe Thr Pro Ala Ala Met Arg Ser Glu 325 330 335 Thr Leu Gly Phe Tyr Pro Trp Lys Pro Thr Ile Pro Thr Pro Trp Arg 340 345 350 Tyr Tyr Phe Gln Trp Asp Arg Thr Leu Ile Pro Ser His Thr Gly Thr 355 360 365 Ser Gly Thr Pro Thr Asn Ile Tyr His Gly Thr Asp Pro Asp Asp Val 370 375 380 Gln Phe Tyr Thr Ile Glu Asn Ser Val Pro Val His Leu Leu Arg Thr 385 390 395 400 Gly Asp Glu Phe Ala Thr Gly Thr Phe Phe Phe Asp Cys Lys Pro Cys 405 410 415 Arg Leu Thr His Thr Trp Gln Thr Asn Arg Ala Leu Gly Leu Pro Pro 420 425 430 Phe Leu Asn Ser Leu Pro Gln Ala Glu Gly Ala Ala Asn Phe Gly Tyr 435 440 445 Ile Gly Val Gln Gln Asp Lys Arg Arg Gly Val Thr Gln Met Gly Asn 450 455 460 Thr Asn Tyr Ile Thr Glu Ala Thr Ile Met Arg Pro Ala Glu Val Gly 465 470 475 480 Tyr Ser Ala Pro Tyr Tyr Ser Phe Glu Ala Ser Thr Gln Gly Ser Phe 485 490 495 Lys Thr Pro Ile Ala Ala Gly Arg Gly Gly Ala Gln Thr Asp Glu Asn 500 505 510 Gln Ala Ala Asp Gly Asp Pro Arg Tyr Ala Phe Gly Arg Gln His Gly 515 520 525 Gln Lys Thr Thr Thr Thr Gly Glu Thr Pro Glu Arg Phe Thr Tyr Ile 530 535 540 Ala His Gln Asp Thr Gly Arg Tyr Pro Glu Gly Asp Trp Ile Gln Asn 545 550 555 560 Ile Asn Phe Asn Leu Pro Val Thr Asp Asp Asn Val Leu Leu Pro Thr 565 570 575 Asp Pro Ile Gly Gly Lys Thr Gly Ile Asn Tyr Thr Asn Ile Phe Asn 580 585 590 Thr Cys Gly Pro Leu Thr Ala Leu Asn Asn Val Pro Pro Val Tyr Pro 595 600 605 Asn Gly Gln Ile Trp Asp Lys Glu Phe Asp Thr Asp Leu Lys Pro Arg 610 615 620 Leu His Val Asn Ala Pro Phe Val Cys Gln Asn Asn Cys Pro Gly Gln 625 630 635 640 Leu Phe Val Lys Val Ala Pro Asn Leu Thr Asn Glu Tyr Asp Pro Asp 645 650 655 Ala Ser Ala Asn Met Ser Arg Ile Val Thr Tyr Ser Asp Phe Trp Trp 660 665 670 Lys Gly Lys Leu Val Phe Lys Ala Lys Leu Arg Ala Ser His Thr Trp 675 680 685 Asn Pro Ile Gln Gln Met Ser Ile Asn Ile Asp Asn Gln Phe Asn Tyr 690 695 700 Leu Pro Ser Asn Ile Gly Gly Met Glu Ile Val Tyr Glu Lys Ser Gln 705 710 715 720 Leu Ala Pro Arg Lys Leu Tyr 725 <210> 6 <211> 2184 <212> DNA <213> Artificial <220> < 223> CPV-2b_embodiment 1_passage 98_nucleotide <400> 6 atggcacctc cggcaaagag agccaggaga ggacttgtgc ctccaggtta taaatatctt 60 gggcctggga acagtcttga ccaaggagaa ccaactaacc cttctgacgc cgctgcaaaa 120 gaacacgacg aagcttacgc tgcttatctt cgctctggta aaaacccata cttatatttc 180 tcgccagcag atcaacgctt tatagatcaa actaaggacg ctaaagattg gggggggaaa 240 ataggacatt atttttttag agctaaaaag gcaattgctc cagtattaac tgatacacca 300 gatcatccat caacatcaag accaacaaaa ccaactaaaa gaagtaaacc accacctcat 360 attttcatca atcttgcaaa aaaaaaaaaa gccggtgcag gacaagtaaa aagagacaat 420 cttgcaccaa tgagtgatgg agcagttcaa ccagacggtg gtcagcctgc tgtcagaaat 480 gaaagagcta caggatctgg gaacgggtct ggaggcgggg gtggtggtgg ttctgggggt 540 gtggggattt ctacgggtac tttcaataat cagacggaat ttaaattttt ggaaaacgga 600 tgggt ggaaa tcacagcaaa ctcaagcaga cttgtacatt taaatatgcc agaaagtgaa 660 aattatagaa gagtggttgt aaataatttg gataagactg cagttaaagg aaacatggct 720 ttagatgata ctcatgcaca aattgtaaca ccttggtcat tggttgatgc aaatgcttgg 780 ggagtttggt ttaatccagg agattggcaa ctaattgtta atactatgag tgagttgcat 840 ttagttagtt ttgaacaaga aatttttaat gttgttttaa agactgtttc agaatctgct 900 actcagccac caactaaagt ttataataat gatttaactg catcattgat ggttgcatta 960 gatagtaata atactatgcc atttactcca gcagctatga gatctgagac attgggtttt 1020 tatccatgga aaccaaccat accaactcca tggagatatt attttcaatg ggatagaaca 1080 ttaataccat ctcatactgg aactagtggc acaccaacaa atatatacca tggtacagat 1140 ccagatgatg ttcaatttta tactattgaa aattctgtgc cagtacactt actaagaaca 1200 ggtgatgaat ttgctacagg aacatttttt tttgattgta aaccatgtag actaacacat 1260 acatggcaaa caaatagagc attgggctta ccaccatttc taaattcttt gcctcaagct 1320 gaaggagctg ctaactttgg ttatatagga gttcaacaag ataaaagacg tggtgtaact 1380 caaatgggaa atacaaacta tatcactgaa gctactatta tgagaccagc tgaggttggt 1440 tatagtgcac catatt attc ttttgaggcg tctacacaag ggtcgtttaa aacacctatt 1500 gcagcaggac gggggggagc gcaaacagat gaaaatcaag cagcagatgg tgatccaaga 1560 tatgcatttg gtagacaaca tggtcaaaaa actaccacaa caggagaaac acctgagaga 1620 tttacatata tagcacatca agatacagga agatatccag aaggagattg gattcaaaat 1680 attaacttta accttcctgt aacagatgat aatgtattgc taccaacaga tccaattgga 1740 ggtaaaacag gaattaacta tactaatata tttaatactt gtggtccttt aactgcatta 1800 aataatgtac caccagttta tccaaatggt caaatttggg ataaagaatt tgatactgac 1860 ttaaaaccaa gacttcatgt aaatgcacca tttgtttgtc aaaataattg ccctggtcaa 1920 ttatttgtaa aagttgcgcc taatttaaca aatgaatatg atcctgatgc atctgctaat 1980 atgtcaagaa ttgtaactta ctcagatttt tggtggaaag gtaaattagt atttaaagct 2040 aaactaagag cctctcatac ttggaatcca attcaacaaa tgagtattaa tatagataac 2100 caatttaact atctaccaag taatattgga ggtatggaaa ttgtctatga aaaatctcaa 2160ctagcaccta gaaaattata ttaa 2184

Claims (11)

For mammalian parvovirus vaccine comprising canine parvovirus type 2b (CPV-2b) or a subculture thereof comprising a VP-2 (viral capsid peptide-2) protein consisting of the amino acid sequence of SEQ ID NO: 3 composition.
The composition for a mammalian parvovirus vaccine according to claim 1, wherein the canine parvovirus type 2b is a live cell or a dead cell.
The composition for a mammalian parvovirus vaccine according to claim 1, wherein the VP-2 protein is encoded by the nucleotide sequence of SEQ ID NO: 4.
The composition for a mammalian parvovirus vaccine according to claim 1, wherein the subculture is canine parvovirus type 2b obtained by subculture of canine parvovirus type 2b 90 to 150 times.
According to claim 1, wherein the subculture is canine parvovirus type 2b comprising a VP-2 protein consisting of the amino acid sequence of SEQ ID NO: 5; or canine parvovirus type 2b of accession number KCTC18847P; A composition for mammalian parvovirus vaccine, characterized in that
The composition for a mammalian parvovirus vaccine according to claim 5, wherein the VP-2 protein comprising the amino acid sequence of SEQ ID NO: 5 is encoded by the nucleotide sequence of SEQ ID NO: 6.
The composition for a mammalian parvovirus vaccine according to claim 1, wherein the mammal is at least one selected from the group consisting of dogs, cats, foxes, wolves, pigs and skunks.
A method for preventing parvovirus infection by administering the vaccine composition according to claim 1 to mammals other than humans.
i) obtaining a sample from the nasal cavity of the mammal;
ii) isolating a parvovirus comprising the VP-2 protein of SEQ ID NO: 3 from the sample of step i); and
iii) subculturing the parvovirus of step ii) 90 to 150 times;
A method for preparing a composition for a mammalian parvovirus vaccine comprising a.
10. The method of claim 9, iv) inactivating the parvovirus by treating the subculture of step iii) with formaldehyde and glycine in a molar concentration ratio of 3:1 to 1:4;
A method for preparing a composition for a mammalian parvovirus vaccine, characterized in that it additionally comprises a.
The method of claim 10, wherein the mammal in step i) is at least one selected from the group consisting of dogs, cats, foxes, wolves, pigs and skunks.

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