RU2242513C1 - Foot and mouth disease virus strain a(georgia)1999/ 1721 type a for preparing diagnostic and vaccine preparations - Google Patents

Abstract

FIELD: biotechnology, virology, veterinary science.

SUBSTANCE: the parent virus for preparing the strain A(Georgia)1999/№1721 is isolated in 1999 year from sick cows in private sector of village Ude in Adigensky district, Georgia Republic. The industrial strain A(Georgia)1999/№1721 is prepared by successive passages in sensitive hetero- and homological cell cultures. The strain is deposited in collection of microorganisms VGNKI at registration number A(Georgia)1999/№1721. Virus of the strain A(Georgia)1999/№1721-DEP is reproduced in sensitive cell cultures with accumulation up to 6.0-8.0 lg TCD₅₀/ml for 18-24 h of incubation. At mass infection virus induces cytopathogenic effect (CPE) in 4 h and retains the parent indices in passage in cell cultures for 10 passages. The strain A(Georgia)1999/№1721-DEP elicits high biological, antigenic and immunogenic activity and can be used for preparing agents for specific prophylaxis and diagnosis of foot and mouth disease virus type A that shows homological properties with epizootic virus distributing in Transcaucasian and Near East countries.

EFFECT: valuable properties of strain.

7 tbl, 3 dwg, 5 ex

Description

The invention relates to the field of veterinary virology and biotechnology and can be used in the manufacture of agents for the specific prophylaxis of the diagnosis of foot and mouth disease type A.

Foot and mouth disease is an acute contagious viral disease of artiodactyl animals, manifested by fever, vesicular (aphthous) lesions of the mucous membrane of the oral cavity, hairless areas of the scalp, udder, corolla, and inter-hoof gap and accompanied by impaired movement. It is characterized by a tendency to widespread and epizootic course. The disease is accompanied by large losses of milk, meat and other types of livestock products, complicates commercial operations and economic activity.

Years of experience show that the presence of endemic foot and mouth disease reduces income in dairy and beef cattle breeding by 30-40%.

The foot and mouth disease virus belongs to the genus of aphthoviruses, the family of picornaviruses. It includes 7 immunological types and a large number of subtypes.

A well-known feature of the causative agent of foot and mouth disease is the antigenic variation of strains within the same serotype, occurring at different time intervals, in different territories, depending on the species composition of the susceptible population, its immune status and many other factors. It is believed that the main cause of antigenic variation is a change in the amino acid sequence in the polypeptides of the viral proteins that make up the virion capsid. In practice, such antigenic changes can be expressed from insignificant differences between the strains, captured only using subtle molecular analysis methods, to the appearance of completely different strains requiring the use of new tools for serological diagnosis and specific prophylaxis of the disease caused by these strains.

Type A foot and mouth disease virus strains have been used as production strains in the USSR for the past 40 years.

These include the following strains: A ₇ No. 103, isolated in 1962 in the Kuibyshev region; A ₇ No. 2, allocated in 1965 in the Tajik SSR; And No. 717/73, allocated in 1973 in the Stavropol Territory.

These strains were used to obtain diagnosticums and FMD vaccines used in various regions of the country.

However, after the elimination of foot-and-mouth disease caused by antigenically similar strains of the virus, they were discontinued and are currently supported only in the Museum of strains of the Federal State Research Institute for the Protection of Animals (1, 2, 4, 5).

The known strain No. 550 of foot and mouth disease virus A ₂₂ , isolated in 1964 in Azerbaijan and used in the Russian Federation as an industrial one in the manufacture of specific prophylaxis and diagnostics used throughout Russia and the CIS countries (6).

The disadvantages of this strain are its low biological, antigenic and immunogenic activity.

This is evidenced by the fact that in 1998 in the Republic of Armenia and in 1999 in the Republic of Georgia foci of FMD were detected in type A cattle immunized with a bivalent AO vaccine, which included antigen from production strain A ₂₂ No. 550. However, the vaccine did not provide satisfactory protection for the immunized animals. Laboratory studies at FGU ARRIAH of aphthous material isolated from sick animals revealed type A foot and mouth disease virus that has antigenic differences from strain A ₂₂ No. 550 in the complement binding reaction and in the virus neutralization reaction. This testified to the inconsistency of the means used for specific prophylaxis and diagnosis of type A foot and mouth disease used by Russia and the CIS countries to the epizootic virus circulating in 1996-1999. in the countries of Transcaucasia and the Middle East (Turkey, Iran).

Closest to the proposed invention in terms of essential features is the strain No. 1707 / Armenia / 98 type A foot and mouth disease virus, isolated on 01.08.98 from sick cows in the farm of Ohchik, Amassi region of the Republic of Armenia. Type A foot-and-mouth disease virus production strain No. 1707 / Armenia / 98 was obtained by repeated consecutive passages on sensitive hetero- and homologous cell cultures. The indicated strain is used for the manufacture of diagnostic and vaccine preparations (7).

The disadvantages of this strain are its antigenic differences from FMD virus isolates isolated in Transcaucasia at different time intervals.

The task of creating the present invention was to obtain a new production strain of foot and mouth disease virus type A, which has high biological, antigenic and immunogenic activity in its native form and after inactivation and provides for the production of diagnostic and vaccine preparations homologous to the epizootic virus that appeared in the countries of the Caucasus and the Middle East.

The technical result from the use of the present invention is to expand the arsenal of production strains of foot and mouth disease virus of serological type A, which have high biological, antigenic and immunogenic activity and are suitable for the manufacture of means for specific prevention and diagnosis of foot and mouth disease type A, homologous to the epizootic virus circulating in the countries of the Caucasus and the Middle East .

The specified technical result was achieved by obtaining strain A (Georgia) 1999 / No. 1721 of the FMD virus type A. Strain A (Georgia) 1999 / No. 1721 is new, previously unknown. The original virus for obtaining strain A (Georgia) 1999 / No. 1721 was isolated in 1999 from sick cows in the private sector of the village of Ude, Adigen region of the Republic of Georgia. Production strain A (Georgia) 1999 / No. 1721 of serological type A was obtained by successive passages on sensitive hetero- and homologous cell cultures.

The resulting strain was deposited in the All-Russian State Collection of microorganism strains used in veterinary medicine and animal husbandry, Federal State Institution "All-Russian State Research Institute for Control, Standardization and Certification of Veterinary Medicines - Quality Center for Veterinary Medicines and Feed" (FGU VGNKI) of the Ministry of Agriculture of the Russian Federation on August 19, 2003 year under registration number A (Georgia) 1999 / No. 1721-DEP.

Strain A (Georgia) 1999 / No. 1721 - DEPT has high biological, antigenic and immunogenic activity in its native form and after inactivation.

The possibility of its use for the preparation of diagnostic preparations and inactivated vaccines has been experimentally confirmed. Strain A (Georgia) / 1999 / No. 1721-DEPT of type A foot and mouth disease virus provides diagnostic products that allow serological diagnosis of type A foot and mouth disease virus isolates causing outbreaks of disease in recent years in the countries of the Caucasus and the Middle East, and anti-FMD inactivated vaccine that protects susceptible animals against the specified pathogen.

The invention is illustrated in the graphic materials, where:

figure 1 presents the amino acid sequence of the protein VP1 strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus serological type A;

figure 2 shows a dendrogram reflecting the phylogenetic relationship of strain A (Georgia) 1999 / No. 1721-DEPH of the virus of foot and mouth disease serological type A with epizootic and vaccine strains of foot and mouth disease type A. The dendrogram is based on a comparison of the complete nucleotide sequences of the VP1 gene;

figure 3 shows the alignment of the amino acid sequences of the protein VP1 strains No. 1707 / Armenia / 98-DEPT and A (Georgia) 1999 / No. 1721-DEPT.

Strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A is characterized by the following features and properties.

Morphological properties

Strain A (Georgia) 1999 / No. 1721-DEPT belongs to the family Picornaviridae, genus Aphtovirus, serotype A and has morphological characteristics characteristic of the causative agent of foot and mouth disease: the form of the virion is icosahedral, size 23-25 nm. Virion consists of an RNA molecule enclosed in a protein coat. The protein shell consists of 32 capsomeres arranged in cubic symmetry.

Antigenic properties

According to its antigenic properties, strain A (Georgia) 1999 / No. 1721-DEPT of foot and mouth disease virus belongs to serotype A.

The virus is stably neutralized by homologous antiserum. The virus does not exhibit hemagglutinating activity. In sick animals, antibodies are formed in the blood serum that are detected in RDP, ELISA and pH. When vaccinating cattle with a vaccine from an inactivated virus, it induces the formation of specific antibodies detected in ELISA, RDP and pH. During hyperimmunization of guinea pigs with a concentrated inactivated virus, it induces the formation of virus-specific antibodies detected in CSC at a dilution of 1: 128-1: 512 and in ELISA at a dilution of 1: 6000-1: 10000.

When determining the antigenic correspondence of strain A (Georgia) 1999 / No. 1721-DEPT with production strains and field isolates of FMD virus type A, isolated during 1962-1999. in the territory of the CIS countries and a number of other states of the world, a comparative study of the primary structure of the VP1 gene and protein was conducted (Fig. 1).

Comparative analysis of the established structures of strain A (Georgia) 1999 / No. 1721-DEPT and epizootic isolates isolated in recent years in Turkey, Iran and the Republic of Armenia (A / Iran / 96, A / Turkey / 97, A / Turkey / 98, A / Armenia / 98), showed that isolate A / No. 1721 / Georgia / 99 is different from isolates A / Turkey / 97, A / Turkey / 98, A / Iran / 96 and A / No. 1707 / Armenia / 98.

Phylogenetic relationships of the studied isolates with previously studied strains isolated in different countries of the world over the past three decades are indicated on the dendrogram (figure 2). It was found that the studied isolates significantly differ from all previously isolated isolates, including the FMD virus strains A ₂₂ No. 550 and A / No. 1707 / Armenia / 98, which are currently used for the production of diagnostics and specific prophylaxis.

The antigenic relationship of strain A (Georgia) 1999 / No. 1721-DEPT with the corresponding production and previously isolated strains in Turkey and Iran was studied in the RSK. The results are presented in table 1.

As shown by the data presented in table 1, strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A differs from both production and previously isolated epizootic strains.

According to the antigenic properties, the new strain A (Georgia) 1999 / No.1721-DEPT of the FMD virus of type A also differs from the previously studied production strain A # 1707 / Armenia / 98-DEPT. Bilateral relationship (R) in the CSC between them is 20%.

A similar study of the antigenic affinity of the isolated strain was carried out in PH, where cattle of convalescents of cattle for viruses of strains A (Georgia) 1999 / No.1721-DEP, A / No.1707 / Armenia / 98, A / Iran / 87 and A _{22 were} used as immune No. 550. The results of these studies are presented in table 2.

The data presented in table 2 confirm the antigenic difference of the isolated strain from production strain A ₂₂ No. 550 and epizootic strains isolated in Iran in 1987 and Uzbekistan in 1989.

Biotechnological characteristics

Strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A exhibits high biological, antigenic and immunogenic activity both in its native form and after inactivation. The strain is intended for the production of diagnostic sera and antigenic drugs, as well as for the manufacture of inactivated FMD vaccines. Strain A (Georgia) 1999 / No. 1721-DEPT is reproduced in monolayer cultures of primary trypsinized culture of pig kidney cells (SP), transplanted cultures of kidney cells of the Siberian mountain ibex (PSGK-30), IB-RS-2, VNK-21, and during 18-24 hours of incubation, up to 6.0-8.0 Ig TCD ₅₀ / ml accumulates. With massive infection (1-100 TCD / cell), it causes a CPD after 4 hours.

After 3-4 incubation passages, up to 6.0-8.0 Ig TCD ₅₀ / ml accumulates. It retains its original characteristics when passaged in sensitive biological systems for 10 passages.

Chemo- and genotaxonomic characterization

Strain A (Georgia) 1999 / No. 1721-DEPT is an RNA-containing virus with a molecular weight of 7 × 10 ⁶ D.

Nucleic acid is represented by a single-stranded linear molecule with a molecular weight of 2.8 × 10 MD. The virion has a protein shell consisting of four basic proteins VP1, VP2, VP3 and VP4. The lipoprotein membrane is absent.

The main antigenic protein is VP1. The virion contains approximately 31.5% RNA and 68.5% protein. Virionic RNA is infectious and is involved in the formation of precursor proteins in infected cells. The precursors, in turn, are cleaved to form more stable structural and non-structural virus polypeptides. Of the 6 non-structural polypeptides that accumulate in infected cells, one (VP3D) is an RNA-dependent RNA polymerase involved in RNA replication of new virions.

The primary structure of the viral genome region was determined using PCR and sequencing. The research results are presented in figure 1.

It was found that strain A (Georgia) 1999 / No. 1721-DEPT has the highest degree of homology of the primary structure of the gene and protein VP1 with isolates Turkey / 99, A / Iran / 99 and A / No. 1707 / Armenia / 98.

The studied isolate differs significantly from production strains. A ₂₂ No. 550 (18% of differences) and A / No. 1707 / Armenia / 98 (20% of differences), as well as other variants and strains of foot-and-mouth disease virus A ₅ , A ₁₀ , A ₁₂ , A / No. 1540 / Uzbekistan / 89 and others (figure 2).

The alignment of amino acid sequences of the VP1 protein of strains A / No. 1707 / Armenia / 98-DEPT and A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A was carried out. The results of the studies are presented in Fig.3.

A comparative analysis shows that the strains differ from each other in seven amino acid residues. Two substitutions (G to R at position 141 and R to H at position 142) are located in close proximity to the RGD site responsible for binding to cell receptors. Numerous studies indicate that for type A foot and mouth disease virus, even single replacements in the structure of the VP1 protein can lead to significant changes in antigenic properties.

Physical properties

The virion mass is 8.4 × 10 ⁻¹⁸ g. A sedimentation coefficient of 146S in the sucrose gradient. Floating density 1.45 g / ml.

Resistance to external factors

Strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A is resistant to ether, chloroform, freon, acetone and other organic solvents and detergents. Most stable at pH 7.0-7.6. PH shifts to both the acidic and alkaline sides lead to inactivation of the virus. It is sensitive to formaldehyde, UV radiation, γ-radiation, high temperatures.

Additional features and properties

Immunogenic activity - immunogen in the composition of an inactivated vaccine.

Reactogenicity - does not possess reactogenic properties.

Pathogenicity - pathogenic for artiodactyl animals, newborn mice, guinea pigs.

Virulence - virulence for naturally susceptible animals with contact, aerosol and parenteral infection.

Stability - retains the original biological properties when passaged in sensitive biological systems for 10 passages.

Based on the data obtained, it can be argued that strain A (Georgia) 1999 / No. 1721-DEPT in terms of antigenic and immunological spectra is an original, taxonomically new, previously unknown variant of FMD virus type A.

To reduce its epizootic danger, it is necessary to ensure timely laboratory diagnosis and vaccination of newly emerging foci of the disease, which requires highly active and specific antibody and antigenic diagnostics and a vaccine obtained using this strain as a production one.

According to the applicant, the proposed strain meets the conditions of patentability "novelty" and "inventive step".

The essence of the invention is illustrated by examples of its use.

Example 1

Getting strain

Strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A was isolated from field material received at ARRIAH as an epithelium of aphthae from cattle suspected of foot and mouth disease during laboratory diagnosis of this disease and its differentiation from other vesicular diseases. When isolating the virus, a complex of biological, virological and biochemical methods was used.

Biological and virological methods included inoculation of cattle field isolate material and subsequent adaptation of the virus to cultures of primary trypsinized and transplanted cell lines. Cell cultures of SP, PSGK-30, IB-RS-2 and BHK-21 were used. Primary and transplantable cell cultures for bioprobe production were grown on the appropriate nutrient media under stationary conditions in 50-100 ml vials, washed from the growth medium and infected with a 10% suspension of aphthous material (the multiplicity of infection delivered 1-10 TCD ₅₀ per cell) prepared in Hanks solution with 0.5% GLA, and antibiotics according to the standard formulation. To remove microflora and ballast cell components, the suspension was treated with chloroform in a ratio of 1:10. After 30 minutes of incubation at 37 ° C, 5–10 ml of the supporting medium was added to the vials and incubated at 37 ° C until the appearance of the viral CPD. In the presence of CPD (rounding of cells, increasing their optical density, degeneration and separation of cells from glass), the bottles were subjected to freezing, thawing, purification of the cell suspension with chloroform and centrifugation at 3000 g for 15 minutes. The resulting virus-containing material was used for subsequent passages and studies in CSCs and ELISA for the presence of a viral antigen, and a set of commercial type-specific serums and serums were used, which were stored in the Museum of Strains ARRIAH.

The results of the adaptation of the virus to various cell cultures are presented in table 3.

The data shown in table 3, indicate good adaptive activity of strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A to the used cell cultures.

The virus isolated using the above methods was studied in CSC with a set of diagnostics for all types of foot and mouth disease virus, vesicular stomatitis, vesicular exanthema and porcine vesicular disease in order to identify the type of accessory and control for purity. The results of virus typing in CSCs are presented in table 4.

The results shown in table 4, the results indicate that the selected virus belongs to type A.

Example 2

Production and testing of hyperimmune serum in guinea pigs

For hyperimmunization of guinea pigs, FMD virus type A antigen of strain A (Georgia) 1999 / No. 1721-DEPT reproduced in a suspension culture of BHK-21 cells is used. The virus-containing suspension is purified from ballast impurities by the addition of 0.05% polyguanidine. The purified virus is concentrated 100 times by adding 10% polyethylene glycol (PEG) m.m. 6000 and inactivate aminoethylethyleneimine (AEEI) at a concentration of 0.01-0.05%, pH 7.8-8.0.

Inactivated antigen concentrate in a mixture with an equal volume of oil adjuvant is administered to guinea pigs in a volume of 1.0 ml intramuscularly. After 21 days, immunization is repeated and after 10 days the animals are bled. Individual serum samples are checked for type specificity and activity in CSCs.

After this, a series is prepared by mixing type-specific individual samples of the same activity. The strain specificity of a serial preparation is determined in one- and two-sided reactions with homo- and heterologous antigens.

After preservation with sodium azide (1: 5000) and holding at + 4 ° C for 30 days, the resulting serum is Packed in 1.0 ml ampoules and subjected to freeze drying.

By the method described in example 2, 2 series of hyperimmune serum were prepared, the characteristics of which are presented in table 5.

The data in table 5 indicate that in the manner described in example 2, diagnostic sera were obtained that meet the requirements for specific activity (8).

Example 3

Obtaining and testing diagnostic antigen

To obtain the antigen for immunochemical reactions, FMD virus of type O strain A (Georgia) 11999 / No. 1721-DEPT adapted to the cell culture of the transplantable PSGK-30 and BHK-21 lines is used. For adaptation, virus-containing material is used in the form of a 10% suspension of cattle aft. Adaptation is carried out for 3-5 consecutive passages. The resulting matrix virus seed is used to produce viral raw materials. Infection of cell cultures and the collection of viral material is carried out according to standard methods. The resulting virus-containing liquid is concentrated 100 times by adding 8-10% PEG. The resulting concentrate is inactivated by heating at 56 ° C for 40 minutes, Packed in ampoules and dried by vacuum sublimation.

The specified method was prepared 2 series of diagnostic antigen, the characteristics of which are given in table 6.

The results of the studies, shown in table 6, indicate that the diagnostic antigens, the specific activity of which meets the requirements (8), were obtained by the method described in example 3.

Example 4

Preparation and testing of inactivated adsorbate vaccine

Inactivated adphrb vaccine against FMD type A is prepared from virus strain A (Georgia) No. 199 / No. 1721-DEPT grown in a suspension culture of BHK-21 cells.

The resulting virus is subjected to inactivation of AEEI at a concentration of 0.025-0.05% at 36-37 ° C for 112-24 hours and a pH of 7.2-7.6. At the end of inactivation, polyhexamethylene guanidine is added to the suspension to a concentration of 0.005-0.007% for flocculation of ballast impurities and inactivation of possible contaminants. The suspension is cooled to 4-8 ° C. After this, sedimentation of flocculated proteins is carried out and their removal. The resulting antigen is subjected to control for avirulence, virus-specific protein content and 146S + 75S virus components, sterility. The required concentration of 146S + 75S components in the vaccine dose of the adsorbed vaccine is obtained by concentration of the antigen with gel of aluminum oxide hydrate (GOA).

For this, the calculated volume of GOA gel of 3% concentration is added to the cooled antigen suspension. After sedimentation of GOA, the calculated volume of the supernatant is drained or the volume of the remaining suspension is measured. The final concentration of GOA should be in the range of 1.62 ± 0.488 P <0.01 mg / ml p = 10, and the concentration of 146S + 75S components of the FMD virus is at least 6.0 μg / ml. Then add a 10% solution of saponin to a final concentration of 0.075%. The resulting vaccine is packaged in glass bottles and subjected to sterility control. The virulence and harmlessness of the vaccine is checked on 5 heads of cattle, introducing the vaccine first under the mucous membrane of the tongue in a dose of 2.0 ml, and then subcutaneously in a dose of 10 ml. Observation of the clinical condition of the animals is 10 days. Avirulent, harmless and sterile vaccine is tested for immunogenic activity in cattle or guinea pigs. The obtained adsorbate vaccine against foot and mouth disease type A has the optimal component composition, μg / ml:

Avirulent and purified

immunogenic material from immunogenic

(146S + 75S) virus components

foot and mouth disease type A

strain A (Georgia) 1999 / No. 1721-DEPT ≥6.0

Gel GOA 1132.0-2108.0

Saponin 750

Supporting Environment 997 142.0-998112.0

The immunogenic activity of one of the production series of the adsorbate vaccine from FMD virus type A strain A (Georgia) 1999 / No. 1721-DEPT was tested for cattle. Her IMD ₅₀ was 0.15 ml, i.e. one ml of the vaccine contained 6.67 imd ₅₀ .

For comparison, the production series of the adsorbate vaccine from foot-and-mouth disease virus A ₂₂ No. 550 had an ImD ₅₀ for KpS equal to 0.22 ± 0.012 ml P <0.001, i.e. in one ml the vaccine contained 4.55 ImD ₅₀ . The volume of the vaccine dose should contain at least 7 ImD ₅₀ .

Example 5

Preparation and testing of inactivated emulsion vaccine

Type A foot-and-mouth disease virus of strain A (Georgia) 1999 / No. 1721-DEPT is grown, inactivated, cleaned from ballast impurities and possible contaminants, and subjected to abirulence, virus-specific protein content, immunogenic components (146S + 75S) and sterility as described in Example 4. The required concentration of 146S + 75S components in a vaccine dose of an emulsion vaccine is obtained by concentration of antigen by flow ultrafiltration or PEG.

For this, BTU 0.5-2 ultrafilters are used. Filtering is carried out under a pressure of 1.5 atm. The resulting antigen concentrate is stored at 4-6 ° C until used in the vaccine. An emulsion vaccine is obtained by dispersing antigen concentrate and oil adjuvant in a ratio of 3: 7-1: 1, respectively, in colloidal mills.

The result is an inactivated emulsion vaccine against foot and mouth disease type A, which is a milk-like liquid insoluble in water.

The vaccine has a liquid consistency, easily resolves from the injection site, does not cause abscesses, does not cause a general reaction in the form of a temperature increase and has pronounced immunogenicity for pigs at a dose of 1 ml 1-21 days after vaccination, for cattle at a dose of 5 ml after 3 -21 days after vaccination and for sheep at a dose of 1 ml 3-21 days after administration. The vaccine is administered intramuscularly to pigs, and cattle and sheep subcutaneously. Inoculated volume must contain at least 2 μg of 146S + 75S components.

The resulting emulsion vaccine against foot and mouth disease type A has the optimal component composition, μg / ml:

Avirulent and purified

immunogenic material from immunogenic

(146S + 75S) foot and mouth disease components

type A strain A (Georgia)

1999 / No.1721-DEPT ≥2.0

Oil adjuvant 500,000.0-700,000.0

Supporting Environment 299998.0-499998.0

The immunogenic activity of the emulsion vaccine was tested on pigs weighing 35-50 kg. The research results are presented in table 7.

The graft volume of 1 ml of the vaccine contains 7.1 ImD ₅₀ for pigs.

Thus, the above information indicates the fulfillment when using the invention of the following set of conditions:

- strain A (Georgia) 1999 / No. 1721-DEPT of FMD virus type A, embodying the invention, is intended for use in agriculture, namely in veterinary virology and biotechnology;

- for the proposed invention in the form described in the independent claim, the possibility of its implementation using the means and methods described in the application or known prior to the priority date is confirmed;

- strain A (Georgia) 1999 No. 1721-DEPT of type A foot and mouth disease virus obtained in accordance with the invention, has a high specific, immunogenic and antigenic activity and is suitable for the manufacture of diagnosticums and inactivated vaccines.

Therefore, the present invention meets the condition of patentability "industrial applicability".

Sources of information

1. Darda P.I. et al. Antigenic properties of foot and mouth disease virus strain A ₂₂ (A _and ). Veterinary Medicine - 1966, 1, 20-22.

2. Bojko A.A. Declaration sur I'apparition en URSS d'un type virus aphteux different des soucfes de type A anterieurement etudies dans le Pays. BOIE, 1965, 64, 2, 1075-1077.

4. Burdov A.N., Dudnikov A.I., Malyarets P.V. and other foot and mouth disease / Ed. A.N. Burdova. - M., Agropromizdat, 1990, 320 p.

5. Syurin V.N. and other viral diseases of animals. - M., VNIITIBP, 1998, 532-548.

6. Instructions for the manufacture and control of a monovalent emulsion vaccine against foot and mouth disease type A, type O, type C, type Asia-1 (from a virus grown in BHK-21 cells). Approved by the GUV of the State Commission of the Council of Ministers of the USSR for Food and Procurement 01.24.91.

7. RF patent No. 2140452; IPC ⁶ C 12 N 7/00, A 61 K 39/135, G 01 N 33/569, A 61 K 39/42, C 07 K 16/08; 10.27.99 g (prototype).

8. Guidelines for the identification and identification of strains of foot and mouth disease virus. Vladimir, 2002.31 p.

Claims (1)

The strain of the virus Apthae epizooticae, fam. Picomaviridae, genus Aphtovirus, type A, collection VGNKI A (Georgia) 1999 / No. 1721-DEPT, for the manufacture of diagnostic and vaccine preparations.

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