Asia-1-type foot-and-mouth disease virus strain for producing of diagnostic and/or vaccine preparations

FIELD: biotechnology, virology.

SUBSTANCE: claimed strain is obtained by passaging of epizootic isolate on sensitive hetero- and homologous cell cultures. Virus strain is reproduced in sensitive cell cultures and after incubation for 18-24 h 6.0-7.66 lg TCD50/ml is accumulated. In case of mass infection strain takes cytopathic effect over 5 hours, and retains starting characteristics for 10 passages.

EFFECT: strain of high biological, antigenic and immunogenic activity.

1 dwg, 7 tbl, 5 ex

 

The invention relates to the field of veterinary Virology and biotechnology, in particular to the strains of FMDV, which can be used as producers of specific antigens in the manufacture of diagnostic and vaccine preparations.

Foot and mouth disease is an acute contagious viral disease of cloven-hoofed animals, manifested by fever, vesicular (aphthous) lesions of the mucous membranes of the mouth, hairless skin of the head, udder, Corolla, Milpitas cracks and accompanied the traffic violation. For this agent tends to be widely distributed and epizootic period. The disease is accompanied by large losses of milk, meat and other animal products, it is difficult for commercial transactions and economic activities. Long experience shows that with endemic FMD the decline in revenues in dairy and beef cattle by 35-40%.

The virus belongs to the genus of aphthovirus, the family of picornaviruses. He has 7 antigenic types and many subtypes (1, 2).

The causative agent of FMD has significant antigenic variability of strains within the same serotype, which is revealed in the various time periods and in different areas and depends on the species composition of susceptible livestock, E. what about the immune status and many other factors. Antigenic variability of the virus of foot and mouth disease caused by substitutions of amino acids in the polypeptide fragments (antigenic epitopes)exposed on the surface of the capsid proteins.

The antigenic shifts the spectrum corresponding to the update of the structure of the new field strain can vary from minor, captured monoclonal antibodies to significant logged using conventional polyclonal immunoglobulins. Significant changes in the antigenic characteristics of the natural strain is likely to cause weakening of specific immunity induced by non-homologous antigen. They also cause difficulties statusbarisvisible diagnosis (2, 3).

In the result there is a need for new diagnostics and specific prophylaxis.

Known strains of FMD virus type Asia-1, which were registered on the territory of the USSR: Armenia in 1973 and 1984, in Tajikistan in 1974, in Turkmenistan in 1978 and in Georgia in 1984 and 2000, while the isolates in these regions was significantly different from the production strain Asia 1 No. 48 of FMD virus serological type Asia-1 (R was 38-58%).

Strain Asia 1 No. 48 was selected in February 1964 on the territory of the Moscow district of the Tajik SSR. Adapted to the body of mice piglets, 1-2-day kralica and Guinea pigs, to culture cells JV for 10-12 passages and accumulated in the title of 7.5 lg LD50/ml; 7,0-8,0 lg LD50/ml; 5,0 lg EID50/ml and 5.0 to 6.0 lg TCD50/ml, respectively. On the antigenic spectrum of this strain differed from the reference variant of this type of "Pakistan" 1/14 and "Israel" 3/63. The specified strain used as production in the manufacture of means of specific prophylaxis and diagnostics used on the territory of the Russian Federation and CIS countries (4).

The disadvantages of this strain are its low biological, antigenic and immunogenic. The vaccine of this strain provides protection against infection with homologous virus.

The closest to the proposed invention, the essential features is the strain Asia 1 No. 1737/Georgia/2000 FMD virus type Asia-1, selected in July 2000 in Bogdanovsky district of the Republic of Georgia from sick cattle (cattle) and obtained as a production by passage isolate the sensitive hetero - and homologous cell cultures. Strain Asia 1 No. 1737/Georgia/2000 is antigenically distinct from the production strain Asia 1 No. 48 and the reaction of complement fixation (R=32%).

The resulting strain deposited 26.01.2001, in the collection of strains of microorganisms UHF VGNKI under the registration name: the virus strain ASU is and serological type Asia-1 No. 1737/Georgia/2000-the DEPOT (5).

Disadvantages strain of the prototype are its low biological, antigenic and immunogenic. The vaccine of this strain provides protection against infection with homologous virus.

In recent years, on the territory of some States in the Middle East, Central Asia and Transcaucasia were recorded outbreak of foot and mouth disease caused by strains of the virus that is different from the production strain Asia 1 No. 48 indicators serological relationship. One of them was dedicated in 2004 in the Republic of Tajikistan.

The FGI ARRIAH laboratory studies aphthous material isolated from sick animals in the Gorno-Badakhshan Autonomous oblast of the Republic of Tajikistan was established FMD virus type Asia-1 having antigenic differences from the production strain Asia 1 No. 48 in RAC (R=27%).

This shows the inconsistency of the means of specific prophylaxis and diagnosis of FMD type Asia-1, used in Russia and CIS countries, epizootic virus circulating in the Republic of Tajikistan in 2004.

In this regard, there is a need to get a new production of the epidemic strain of FMD virus serological type Asia-1 to ensure the security of Russia and neighboring countries from this agent.

In task C is of the present invention was to obtain a new production strain of FMD virus serological type Asia-1, possessing high biological, antigenic and immunogenic activity in the native form and after inactivation and fabricate diagnostic and vaccine homologous epizootic virus that emerged in the countries of Central Asia.

The technical result from use of the present invention is to expand the Arsenal of industrial strains of FMD virus serological type Asia-1, with high biological, antigenic and immunogenic activity and suitable for the manufacture of means of specific prophylaxis and diagnosis of FMD type Asia-1, homologous epizootic virus circulating in the countries of Central Asia.

This technical result is achieved by obtaining strain Asia 1 /Tajikistan/2004(1960) of FMD virus type Asia-1 (author's name) for the manufacture of a diagnostic and/or vaccines. The strain is new, previously unknown. The original virus to obtain strain Asia 1/Tajikistan/2004(1960) selected in January 2004 from sick cows in the Gorno-Badakhshan Autonomous region of Tajikistan. Production strain Asia 1/Tajikistan/2004 (1960) of FMD virus type Asia-1 obtained by passirovannym epizootic isolate the sensitive hetero - and homologous cell culture.

The resulting strain deponirawe the August 11, 2004 at the Russian state collection of strains of microorganisms, used in veterinary medicine and animal production, Federal state institution "all-Russian state research Institute for control, standardisation and certification of veterinary preparations Center quality of veterinary drugs and feed" (FGI VGNKI) under the registration number (reference): a strain of FMDV Asia 1/Tajikistan/2004 (1960)-DEPT.

Compared with the prototype strain Asia 1/Tajikistan/2004 (1960) has a higher biological, antigenic and immunogenic activity in the native form and after inactivation. Experimentally confirmed the possibility of its use for the preparation of diagnostic products and inactivated vaccines. Strain Asia 1 /Tajikistan/2004 (1960) provides the receive diagnostic products, allowing for serological diagnosis of isolates of FMD virus type Asia-1, causing outbreaks in recent years in the countries of Central Asia, and inactivated FMD vaccines that creates the protection of susceptible animals against a specific pathogen.

The invention is explained on the dendrogram (see drawing)showing the phylogenetic relationships of strain Asia 1/Tajikistan/2004 (1960) with epidemic and vaccine strains of FMD virus type Asia-1 (dendrogram based on the comparison paniculately sequences of the VP1 gene), and in the sequence listing, in which:

SEQ ID N 0:1 is the nucleotide sequence of the gene VP1 strain Asia 1 /Tajikistan/2004 (1960) of FMD virus type Asia-1;

SEQ ID N 0:2 is the amino acid sequence of VP1 strain Asia 1 /Tajikistan/2004 (1960) of FMD virus type Asia-1.

Strain Asia 1/Tajikistan/2004 (1960) of FMD virus type Asia-1 is characterized by the following characteristics and properties.

Morphological properties

Strain Asia 1/Tajikistan/2004 (1960) of FMD virus type Asia-1 belongs to the family Picornaviridae, genus Aphtovirus, serotype Asia-1 and has morphological features that are specific to the causative agent of foot and mouth disease: a form of icosahedral virion, size 23-25 nm. The virion consists of a molecule of RNA enclosed in a protein shell. The protein shell consists of 32 capsomeres located in the cubic symmetry.

Antigenic properties

According to their antigenic properties of the strain Asia 1/Tajikistan/2004 (1960) refers to serotype Asia-1. The virus is stable neutralized homologous anticorodal. The virus does not manifest hemagglutinin activity (HA activity). The animals recover in the blood serum of antibodies are formed, detected in the reaction diffusion precipitation (RDP), the enzyme-linked immunosorbent assay (ELISA) and neutralization (PH). When vaccination of cattle vaccine of inactivated virus induces the formation of specificeskich antibodies detected in ELISA, RDP and PH. If hyperimmunization Guinea pigs concentrated inaktivirovannye antigen induces education virousspecificakih antibodies detected in the reaction of complement fixation (RAC) in dilutions 1:128-1:180 and ELISA at dilutions of 1:4000-1:6000,

Antigenic relatedness of strain Asia 1/Tajikistan/2004 (1960) with a production strain Asia 1 No. 48 and strains, previously isolated in Iran and Transcaucasia, studied in RAC. The results of these studies are presented in table 1.

The data in table 1 indicate that the strain Asia 1/Tajikistan/2004 (1960) differ from the production strain Asia 1 No. 48 (R=27%), from strain Asia 1 No. 1737/Georgia/2000 (R=42%), as well as from previously allocated epizootic strains of FMD virus type Asia-1. At the same time he is antigemmoroidalnym strains Asia-1 No. 1238/Georgia/84 and Asia-1/Pakistan 1/54.

Molecular-genetic characteristics

Method of nucleotide sequencing determined the primary structure of the gene VP1 (SEQ ID N 0:1) strain Asia 1/Tajikistan/2004 (1960) and from it is deduced amino acid sequence of VP1 protein (SEQ ID N 0:2).

A comparative analysis of the set of sequences showed that the primary structure of the gene and protein VP1 strain Asia 1/Tajikistan/2004 (1960) differs significantly from the Russian vaccine strain Asia 1 No. 48 and has close phylogenetic is some kinship with the epidemic isolates Iran/99, Georgia/2000, Georgia/2001 Armenia/2000. Phylogenetic relationships of strain Asia 1/Tajikistan/2004 (1960) with the previously studied isolates of FMD virus type Asia-1, isolated in different regions of the world over the past four decades, reflected in the dendrogram.

Biotechnological characteristics

Strain Asia 1/Tajikistan/2004 (1960) of FMD virus type Asia-1 shows high biological, antigenic and immunogenic activity as in the native form and after inactivation.

The FMD virus strain Asia 1/Tajikistan/2004 (1960) reproducerea in monolayer cell cultures of kidney pig (SP), transplantable cell cultures kidney Siberian mountain goat (PSGC-30) and IB-RS-2 for 18-24 hours, accumulating in the credits from 6.0 to 7,66 lg TCD50/ml. At high multiplicity of infection (1-10 TPD/cell) causes the JRC in 5 hours. Preserves the original characteristics when passirovannye in cell cultures for 10 passages (observation period).

The strain intended for diagnostic sera, antigen preparations, and also for the manufacture of inactivated FMD vaccines.

Chemo - and geotectonically feature

Strain Asia 1/Tajikistan/2004 (I960) is an RNA-containing virus with a molecular mass of 7×106D.

Nucleic acid represented by single-stranded linear mole who uloi molecular weight 2,8× 106D. the virion has a protein shell composed of four basic proteins VP1, VP2, VP3 and VP4. Lipoproteina shell is missing.

The major antigenic protein VP1 is. The virion contains approximately 31.5% of RNA and 68.5% protein. Firiona RNA is infectious and is involved in the formation of protein precursors in infected cells.

In turn, the precursors are decomposed with the formation of a more stable structural and non-structural proteins of the virus. Of the 6 non-structural polypeptides that accumulate in infected cells, one (VP66A) is an RNA-dependent RNA polymerase that is involved in the replication of RNA new virions.

Physical properties

The mass of the virion is 8.4×10-18, the sedimentation Coefficient 146S and sucrose gradient. Floating density of 1.45 g/ml

Resistance to external factors

The virus strain Asia 1/Tajikistan/2004 (1960) resistant to ether, chloroform, freon, acetone and detergents. Most stable at pH 7,2-7,6. Changes of pH in acidic and in the alkaline side lead to inactivation of the virus. Sensitive to formaldehyde, UV-irradiation, γ-radiation, and high temperatures.

Additional characteristics and properties

Immunogenic activity - immunogene comprising inactivated vaccine.

Reactogenicity - reactive properties not on the fully.

The pathogenicity of the pathogenic for cloven-hoofed animals, newborn mice, rabbits, Guinea pigs.

Virulence - virulent for naturally-susceptible animals in contact, aerosol and parentelem infection.

Stability - maintains the original biological properties when passirovannye in sensitive biological systems within 10 passages (observation period).

Based on the obtained data, it can be argued that the strain Asia 1 /Tajikistan/2004 (1960) antigenic and immunological spectra is original, in taxonomic relation to new, previously unknown variant of FMD virus type Asia-1.

To reduce its epizootic risk need to provide timely laboratory diagnosis and prevention of emerging outbreaks. Which requires a highly active and specific and antigenic or antibody-based test diagnostics and vaccine derived from the use of this strain as a production.

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

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

Example 1.

Strain Asia 1/Tajikistan/2004 (1960) of FMD virus type Asia-1 was isolated from field material (examinations the No. 1960), received 23.01.2004, FGI ARRIAH of the Gorno-Badakhshan Autonomous oblast of the Republic of Tajikistan in the form of the epithelium aft from cattle suspected of disease foot and mouth, when the laboratory diagnosis of this disease and its differentiation from other vesicular diseases. When isolation of the virus used the complex biological, virological and biochemical methods stipulated by the guidelines for the detection and identification of strains of FMDV. Vladimir, 2002, 31 S.

These methods included intradermically inoculation material field isolates of cattle (1 passage) and the subsequent adaptation of the virus isolated from a diseased animal to cultures of primary and transplantable cell lines. We used cell culture JV, PSGK-30, IB-RS-2 and KSS-21. Primary and transplantable cell culture for the production of assay were grown in appropriate culture media in bottles with a capacity of 50 ml, washed from the growth medium and used to infect 10% suspension aphthous material (multiplicity of infection was 1-10 TCD50for a cell), prepared in Hanks solution with 0.5% GLA and antibiotics according to the standard recipe. Removal of microflora and ballast cellular components, the suspension was treated with chloroform (10%). After a 30 minute incubation at +37°With vials made is about 5-10 ml of maintenance medium and incubated at +37° With until JRS virus. In the presence of JRS (rounding of cells, increasing their optical density, degeneration and separation of the cells from the glass vials were subjected to freeze-thawing, cleaning the cell suspension chloroform and centrifugation at 3000 g for 15 minutes. Received vaccinated material used for the subsequent passages and research at the RAC for the presence of viral antigen, used a set of commercial typespecification sera and sera stored at the Museum of strains FGI ARRIAH.

The results of the adaptation of the virus to different cell cultures are presented in table 2.

The data in table 2 indicate a good adaptive activity of the strain Asia 1/Tajikistan/2004(1960) used cell cultures.

Isolated using the above methods virus preparation was investigated in RAC with a set of diagnostics on all types of FMD virus and vesicular stomatitis to identify typical facilities and control of purity. The results of typing of the virus in RSK are shown in table 3.

The data in table 3, indicate that the material examination No. 1960 identified complimentative antigen of FMDV type Asia-1 at a dilution of 1:2.

Example 2.

For hyperimmunization Guinea pigs using an the Egan from strain Asia 1 /Tajikistan/2004(1960) of FMD virus type Asia-1, obtained in suspension culture cells KSS-21. Vaccinated suspension clear of ballast impurities by the addition of 0.05% solution of guanidine (phmg). The purified virus is concentrated 100 times by adding 10% of polyethylene glycol (PEG) 6000 mm and inactivate aminoethylethanolamine (AAAI) at a concentration of 0.01 to 0.05%, pH 7.8 to 8.0.

Inactivated concentrate antigen mixed with an equal volume of oil adjuvant injected Guinea pigs in a volume of 1.0 ml intramuscularly. After 21 day of immunization and repeat after 10 days, animals bleed. Individual samples of blood serum test sample specificity and activity in the CBD. After that prepare a series by mixing typespecification individual serum samples of the same activity. Strain specificity of serial drug determine in single and duplex reactions with Homo - and heterologous antigens.

After canning sodium azide (1:5000) and incubation at 14°C for 25-30 days the resulting serum is Packed in ampoules of 0.5 to 1.0 ml.

Was prepared 1 series hyperimmune serum, the characteristics of which are presented in table 4.

The data in table 4 indicate that the received diagnostic serum, specific activity meets the requirements of GOST 25384-82.

Example 3.

For the Holocene antigen for immunochemical reactions using the FMD virus type Asia-1 strain Asia 1/Tajikistan/2004 (1960), adapted to the cultures of transplantable cells lines PSGC-30 or VPK-21.

For adaptation used vaccinated material in the form of a 10% suspension of the aft cattle. Adaptation can be performed during 3-5 consecutive passages. The obtained matrix seed is obtained virus used for producing viral material. Contamination of cell cultures and collection of viral material is conducted according to conventional methods. Received vaccinated liquid concentrated 100-fold by adding 8-10% PEG. The obtained concentrate inactivate AAAI, Packed in vials and dried by sublimation under vacuum.

In this way was prepared 2 series diagnostic antigen, the characteristics of which are given in table 5.

The results of the studies are shown in table 5, indicate that the diagnostic antigen-specific activity which meets the requirements of GOST 25384-82.

Example 4.

For the manufacture of the adsorbate-vaccine FMD virus type Asia-1 strain Asia 1 /Tajikistan/2004 (1960) reproduce in suspension culture cells VPK-21.

As a supportive environment using the solution of the Earl without serum with the addition of FGMS, GBX and antibiotics at pH 7.4 and 7.6. Culture of cells infected by a virus from the calculation of 0.01-0.1 TCD50on the cell.

The virus cultivation is carried out at a temperature of 35-36,5°C. H is cut 8-10 hours of incubation shall count live and dead cells at colouring Trifanova blue. If the number of living cells is 15-20%, the incubation continued for another 1-3 hours. When you reach the number of dead cells 90-95% cultivation cease and vaccinated suspension control for sterility and content 146S+75S components. The number 146S+75S components in the suspension should be at least 0.5 μg/ml at the end of the cycle of reproduction of the virus, without temperature control, in vaccinated suspension add 15-20% solution AAAI, acidified with ice-cold acid to a pH of 8.0 to 8.5. The final concentration AEEI in vaccinated suspension must be equal 0,025-0,05%. Inactivation of the infectivity of the virus is carried out in 12-24 hours at 36-37°and a pH of 7.2 and 7.6 with stirring over 5-6 hours for 3-5 minutes. After inactivation of the antigen suspension is cooled to 4-8°C. To the cooled suspension is added 10% solution of pgmg to the concentration of 0,005-0,007% for ballasted flocculation of impurities and inactivation of possible contaminants. Flocculated ballast impurities is subjected to sedimentation, followed by decantation. The resulting antigen control for avirulence, content virousspecificakih protein and 146S+75S components of the virus, sterility. The necessary concentration of 146S+75S components in pravilnoy dose adsorbed vaccine is obtained by concentration of the antigen by gel hydroxide and uminia (GOA).

To a cooled suspension of antigen when operating the mixer add the calculated volume of the gel GOA 3% concentration. Mixing lead within 30 minutes. After sedimentation of the gel GOA merge estimated volume of the supernatant liquid or measure the volume of the remaining suspension. The final concentration of GOA should be in the range of 1.62+0,488 mg/ml, p<0,01 n=10, and the concentration 146S+75S components of FMD virus is not less than 2.0 µg/ml Then in the slurry, add an additional 10% solution of saponin to a final concentration of 0.075%. The resulting vaccine is filled into glass vials and conduct monitoring sterility according to GOST 28085-89 "biological Preparations. Method of biological control of sterility".

The avirulence and safety of the vaccine tested for 5 heads of cattle, introducing the vaccine first, under the mucous membrane of the tongue at a dose of 2 ml, and then subcutaneously at a dose of 10 ml of monitoring the clinical condition of the animals are 10 days. Avirulent, harmless and sterile vaccine tested for immunogenic activity in cattle or Guinea pigs.

The resulting vaccine is a liquid light amber with a loose white solid sorbent, which is formed on the bottom of the vial during storage and can be easily broken in a homogeneous suspension with shaking.

Table 6 shows the results of comparative experiments on the cultivation of FMD virus strain Azi is-1/TJ/2004(1960) and virus production strain Asia 1 No. 48 in the cultivators of the COP-6, from which it follows that the FMD virus strain Asia 1/Tajikistan/2004(1960) has a shorter period of optimal reproduction that necessitated the selection of the optimal conditions for the cultivation of the virus and multiplicity of infection.

Example 5.

Comparative tests were performed on Guinea pigs immunogenic FMD vaccine, inactivated, adsorbed from strain Asia 1/Tajikistan/2004(1960), and FMD vaccine, inactivated, adsorbed from the production strain Asia 1 No. 48.

To determine the immunogenic activity of each drug used in 40 Guinea pigs, of which 8 were control.

The vaccine was diluted in phosphate buffer solution in a ratio of 1:3, 1:9, 1:27 and 1:81. For each dilution of the vaccine took 8 Guinea pigs. Each group of 8 Guinea pigs were kept in a separate cage, which was assigned the label indicating series of the vaccine, its cultivation, the date of commencement of trials and the number of Guinea pigs vaccinated this breeding.

Each vaccine was administered to 4 groups of Guinea pigs intramuscularly in the volume of 2 ml in the region of the right and left femur in 1 ml Control group of Guinea pigs were used to test the activity of the control serotypes of FMD virus.

For vaccinated pigs were observed for 15-17 days. On the Le of this all vaccinated and control animals were infected adapted to them the virus of homologous and heterologous strains intradermal in the plantar surface of one of the hind legs at a dose of 10 -4GD50/0,1 ml.

Analysis of infection was performed at 3, 5 and 7 days and was assessed by generalization yasunaga process on the front and one hind legs.

Control vaccine was considered valid if 8 control pigs generalized form of foot-and-mouth disease for at least 7 animals.

Serum from Guinea pigs, selected 30 days after vaccination were tested for the presence of neutralizing antibody in the reaction of microneutralization (RMN) with homologous and heterologous strains.

The results of these studies are presented in table 7, indicate that samples of sera from Guinea pigs immunized with FMD vaccine from strain Asia 1 No. 48, RMN with the homologous strain, detect higher antibody titers and less low with heterologous strain Asia 1/Tajikistan/2004(1960).

Serum from Guinea pigs immunized with FMD vaccine, inactivated adsorbed from strain Asia 1/Tajikistan/2004(1960), reveal a high antibody titers in RMN as with the homologous strain and heterologous.

Thus, the above information shows the implementation of the use of this invention the following cumulative conditions:

a strain Asia 1/Tajikistan/2004(1960) of FMD virus type Asia-1 embodying th is my invention, designed for use in agriculture, namely in veterinary Virology and biotechnology;

for the present invention in the form as it is described in the independent claim, confirmed the possibility of its implementation using the steps described in the application or known before the priority date tools and methods;

a strain Asia 1/Tajikistan/2004(1960) of FMD virus type Asia-1, obtained in accordance with the invention, it has high biological, specific, antigenic and immunogenic activity in the native form and after inactivation and suitable for the manufacture of a diagnostic and/or vaccines.

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

Sources of information

1. Rarer X. FMD. TRANS. with it. Gasartobi, Ed. and Annot. Kida. wet. Sciences Pivalate. - M., spike, 1971, 432 S.

2. Burdov A.N., Dudnikov A.I., Malaret PV and other FMD /edited Angov. - M, Agropromizdat, 1990, 320 S.

3. Syurin NR. and other Viral diseases of animals. - M, VNITIBP, 1998. - S-548.

4. Instruction for production and control of vaccines monovalent emulsion against FMD type a, type O, type C, type Asia-1 (from virus grown in cell VPK-21). Approved BS of the state Commission of the USSR food and is acwpcum 24.01.91.

5. RF patent №2218397; 12N 7/00, AK 39/135 // (12N 7/00, 12R 1:93); 10.12.2003, (prototyp).

Table

Antigenic spectrum of strain Asia 1/Tajikistan/2004(1960) of FMD virus type Asia-1 according to the RSK
Compare strainsIndicators
r1r2R%
Asia-1 no 480,170,4627
Asia-1 No. 1737/Georgia/20000,390,4642
Asia-1 No. 1730/Iran 58/990,160,0913
Asia-1 No. 1731/Thailand 2/950,560,2537
Asia-1 No. 746/Tajikistan/740,241,049
Asia-1 No. 1238/Georgia/840,780,6470
Asia-1/Pakistan 1/540,820,7076
Asia-1/Iran 1/730,500,76 61
Asia-1 No. 674/Armenia/730,660,7168
Table 2

Biological properties of strain Asia 1/Tajikistan/2004(1960) of FMD virus type Asia-1
The system of reproduction of the virusThe time of manifestation of biological activity (hours)The number of adaptive passagesCharacteristics of the adapted material
Activity in RACThe titer of infectivity lg TCD50/ml
SP18-963one piece.-1:43,0-6,0
PSGC-3018-1203one piece.-1:42,5-6,33
IB-RS-218-723one piece.-1:84,5-7,66
KSS-211811:128,25

Table 3

Determining the type of virus in 33%suspension aphthous material (examination No. 1960/Tajikistan/2004)
g/and whey in your breedingExamination No. 1960Control
solid1:21:41:81:16And22O1With1Asia-1SAT-1SAT-2The SAT-3Sun IndianaSU NDK/s
A22No. 550------4---------
O1No. 194-------4--------
With1No. 564---- ----4------
Asia-1 no 4844-------4------
CAT-1 No. 96----------4-----
CAT-2 No. 325-----------4----
CAT-3--- ---------4---
Sun Indiana-------------4--
VS new Jersey--------------4-
Without saw.-----to/a----------
Note: 4-100% delayed hemolysis;

Table 4

Characteristic diagnostic sera obtained on the antigen from strain Asia 1/Tajikistan/2004/(1960) of FMD virus type Asia-1
Name of productVolume, mlActivity in RAC
with homologous diagnosticumwith the heterologous diagnostics Asia-1
Hyperimmune serum1001:80No. 48No. 1737No. 1238Asia-1 Pakistan 1/54
1:201:201:501:50

The virus strain Aphtae epizooticae, .Picomaviridae, genus Aphtovirus, type Asia-1, collection of the FGI " VGNKI " Asia-1/TJ/2004(1960)-the DEPOT, for the manufacture of a diagnostic and/or vaccines.



 

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9 dwg, 1 tbl, 2 ex

FIELD: virology, medicine.

SUBSTANCE: claimed strain is obtained by genetic reassortment of A/Wyoming/3/03 (H3N2) epidemic virus with cold adapted A/Leningrad/134/17/57 (H2N2) strain, which represents attenuation donor, followed by selection in presence of serum for A/Leningrad/134/17/57 (H2N2) virus. Obtained strain is temperature sensitive and cold adapted. Reassortant has taken over epidemic virus two genes encoding hemagglutinine and neuraminidase surface proteins and six genes encoding non-glycosilated proteins from attenuation donor.

EFFECT: new strain for production of living influenza intranasal vaccine for infants and adults.

1 dwg, 3 tbl, 2 ex

FIELD: veterinary virology.

SUBSTANCE: invention relates to 5 strains of II type porcine circovirus (PCV II) which represents causative agent of porcine post-wealing multy-systemic wasting syndrome (PMWS). Disclosed are various immunogenic compositions and vaccine based on said strains for PMWS prophylaxis and/or treatment. Also disclosed are vectors, viral preparations, cell extracts, cell culture supernatants, containing PCV II or nucleotide or protein components thereof; method for PCV II diagnosis, as well as diagnostic composition and kit.

EFFECT: new agent for treatment of porcine PMWS.

178 cl, 7 dwg, 5 tbl, 19 ex

FIELD: veterinary virology, biotechnology.

SUBSTANCE: claimed vaccine contains antigenic material from virus strain of bird infectious bursal disease (IBD), ethylene imine as inactivation agent; and oily adjuvant in effective ratio. Claimed strain is reproduced on 9-11-days embryos of SPF-hens with infective activity of 5.25-7.0 lg EID50/0.2 cm3. Vaccine represents homogenous white emulsion containing inactivated antigen of IBD virus emulsified in oily adjuvant and is administered to healthy bird being 110-120 days old in volume of 0.5 cm2, in single dose. Vaccine of present invention induces high levels of antibodies against IBD of parent bevy and provides anchored passive immunity in 15-25-day young birds.

EFFECT: safe vaccine against bird infectious bursal disease.

8 cl, 5 tbl, 6 ex

FIELD: veterinary virology, biotechnology.

SUBSTANCE: claimed vaccine contains antigenic material from virus strain of bird infectious bursal disease (IBD), ethylene imine as inactivation agent; aluminum hydroxide and saponin as adjuvant in effective ratio. Claimed strain is reproduced on 9-11-days embryos of SPF-hens with infective activity of 5.25-7.0 lg EID50/0.2 cm3. Vaccine represents rose-brown liquid and is administered to healthy bird being 110-120 days old in volume of 0.5 cm2, in single dose. Vaccine of present invention induces high levels of antibodies against IBD of parent bevy and provides anchored passive immunity in 15-25-day young birds.

EFFECT: safe vaccine against bird infectious bursal disease.

13 cl, 5 tbl, 6 ex

FIELD: veterinary virology, biotechnology.

SUBSTANCE: the suggested vaccine contains avirulent and purified antigenic material out of strain A (Georgia) being homologous to infection agent 1999/N1721-DEP obtained in passaged cell culture VNK-21 being the suspension that contains, predominantly, 146S and 75S immunogenic components of foot-and-mouth disease virus, adjuvants aluminum hydroxide with saponin and maintenance medium in efficient ratios. The vaccine is of high immunogenicity and is capable to provide efficient protection against homologous infection agent circulating in Transcaucasian countries and those of Central Asia, Near and Middle East.

EFFECT: higher efficiency.

10 cl, 1 dwg, 4 ex, 10 tbl

FIELD: veterinary virology, biotechnology.

SUBSTANCE: the suggested vaccine contains avirulent and purified antigenic material out of strain A (Georgia) being homologous to infection agent 1999/N1721 obtained in passaged cell culture VNK-21 being the suspension that contains, predominantly, 146S and 75S immunogenic components of foot-and-mouth disease virus. Moreover, the vaccine contains maintenance medium and butyric adjuvant in efficient ratios. The strain has been deposited in collection of FGU VGNKI under registration number - industrial culture strain of foot-and-mouth disease virus A (Georgia) 1999/N1721-DEP of serotype A. As maintenance medium it is necessary to apply serum-free Earle's solution at addition of FGMS, GBCS and antibiotics at pH being 7.4-7.6. Out of butyric adjuvants the vaccine contains butyric adjuvant of the All-Russia Research Institute of Animal Protection (VNIIZZH) or butyric adjuvant of Montanide ISA-70 or Montanide ISA-260 marks by "Seppic" (France). The vaccine provides efficient protection against homologous infection agent circulating in Transcaucasian countries and those of Central Asia, Near and Middle East.

EFFECT: higher efficiency.

11 cl, 1 dwg, 5 ex, 8 tbl

The invention relates to veterinary Virology and biotechnology

The invention relates to the field of veterinary Virology and biotechnology

The invention relates to veterinary Virology and biotechnology

The invention relates to veterinary Virology and biotechnology

The invention relates to biotechnology

FIELD: veterinary virology, biotechnology.

SUBSTANCE: the suggested vaccine contains avirulent and purified antigenic material out of strain A (Georgia) being homologous to infection agent 1999/N1721 obtained in passaged cell culture VNK-21 being the suspension that contains, predominantly, 146S and 75S immunogenic components of foot-and-mouth disease virus. Moreover, the vaccine contains maintenance medium and butyric adjuvant in efficient ratios. The strain has been deposited in collection of FGU VGNKI under registration number - industrial culture strain of foot-and-mouth disease virus A (Georgia) 1999/N1721-DEP of serotype A. As maintenance medium it is necessary to apply serum-free Earle's solution at addition of FGMS, GBCS and antibiotics at pH being 7.4-7.6. Out of butyric adjuvants the vaccine contains butyric adjuvant of the All-Russia Research Institute of Animal Protection (VNIIZZH) or butyric adjuvant of Montanide ISA-70 or Montanide ISA-260 marks by "Seppic" (France). The vaccine provides efficient protection against homologous infection agent circulating in Transcaucasian countries and those of Central Asia, Near and Middle East.

EFFECT: higher efficiency.

11 cl, 1 dwg, 5 ex, 8 tbl

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