Method for making inactivated emulsified avian influenza vaccine and inactivated emulsified avian influenza vaccine

FIELD: medicine; veterinary science.

SUBSTANCE: method involves preparation of antigen material from avian influenza virus strain thereafter cleaned from ballast impurity, virus inactivation, mixing of antigen material and oil adjuvant with controlling the end product. The avian influenza virus strain is represented with the strain "Primorsky" of avian influenza virus referred to Orthomyxoviridae family, Influenzae virus avicum type of serotype A, subtype H5N2, collection Federal State Institurion VGNKI, serial №129 - "ДЕП". The vaccine made under the given method, contains cleaned and avirulent antigen material from strain " Primorsky" and oil adjuvant in ratio (wt %): antigen material - 30.0-40.0, oil adjuvant - 60.0-70.0.

EFFECT: decreased labour and power inputs in making the vaccine and improved quality of antigen material, high antigen activity of the vaccine and effective protection of susceptible poultry from epizootic virus of subtype H5.

14 cl, 5 tbl, 6 ex

 

Group of inventions relates to veterinary Virology and biotechnology and can be used in the manufacture of tools for the specific prevention of avian influenza, caused by a strain of H5N2.

Avian influenza is OSTROBRAMSKA infectious disease of chickens viral etiology, cause significant economic damage to the development of poultry industry in many countries of the world.

Virus H5N2causes a disease of young and adult birds of all kinds. The disease can occur in different forms and in varying degrees of severity. The waterfowl disease usually occurs in the intestinal form of mild severity. Other sensitive birds is the most common respiratory syndrome from inapparent fatal. In General, the virus is characterized as epitheliotropic, but some strains can cause generalized form of the disease with the defeat of almost all organs and systems.

Vaccine prevention of influenza birds occupies a leading place in the fight against this disease.

Known methods of producing vaccines against avian influenza through the use of genetic engineering techniques and vaccines obtained by these methods (see, for example, US 5916879 And, 29.06.1999 or FR 2751225 And 23.01.23). These vaccines are highly effective, but the method of their production is very time-consuming.

Known the s a method of manufacturing inactivated emulsified vaccine against bird flu and the vaccine inactivated emulsified against bird flu, contains the active substance in the form of antigenic material from strain A/Chicken/queretaro/14588-19/95 (H5N2) influenza virus in birds, received a 10-day-old embryos SPF chickens with infectious activity of 7.3 lg EDS50/1 ml and hemagglutinine activity 1:64, cleared of ballast impurities and inactivated with 2-bromethalin hydrobromide and the target additive in the form of an oil adjuvant at a ratio of 1:4 (see, for example, Swayne D.E. et al., Influents of virus strain and antigen mass on the efficacy of H5 avian influenza inactivated vaccines, Avian Pathol., 1999, 28, 245-255).

Most loved ones are the method of manufacture of inactivated emulsified vaccine against bird flu and the vaccine inactivated emulsified against bird flu, containing the active substance in the form of antigenic material from strain A/ Turkey/Minnesota/3689-1551/81(H5N2) influenza virus in birds, received a 10-day-old embryos SPF chickens with infectious activity of 8.7 lg EDS50/1 ml and hemagglutinine activity 1:1024, cleared of ballast impurities and inactivated with formalin, and the target additive in the form of an oil adjuvant in the effective ratio (see, for example, Swayne D.E. et al., Efficacy of vaccines in Chicken against Patogenic Hong Kong H5N1 Avian Influenza, Avian Dis, 2001, 45, 2, 355-365).

Technology of production of inactivated vaccines against avian influenza has drawbacks (for example, to obtain antigenic material use only what about chorioallantois liquid, received after the reproduction of the virus in chicken embryos, in which the antigen content often varies), the elimination of which remains a challenge and is the main prerequisite for studies on its improvement and protection of the poultry industry of our country from this dangerous disease.

In the task of creating the present invention was to develop a method of manufacture of inactivated vaccine and receiving influenza vaccine birds using non-pathogenic strains with the antigenic formula H5N2 that guarantees the protection of bird flu, caused by serovariants H5, without endangering the worsening epidemic situation, and using the test systems on neuraminidase to differentiate antibodies to field strains from the vaccine. This vaccine should have a high immunogenic activity and emissions and to protect the poultry industry of the Russian Federation from virus GP, causing significant economic damage.

The technical result from the use of the invention is to increase the antigenic and immunogenic activity and safety of inactivated vaccines against avian influenza.

The task in terms of method of manufacture of inactivated emulsified vaccine against bird flu is solved in that in the method, including the preparation and digennaro material from a strain of the influenza birds purification of antigenic material from the ballast impurities, the inactivation of the virus, the connection antigenic material with an oil adjuvant and control of the target product, according to the invention, as a strain of the influenza birds use the strain "seaside" influenza of birds belonging to the family Orthomyxoviridae, genus Influenzae virus avicum, serotype a, subtype H5N2, a collection of FGI VGNKI, registration No. 129-DEPT.

To obtain the antigenic material used carcass 11-day-old chick embryos infected with the pathogen.

Antigenic material is used in the form vaccinated suspension, made of extraembryonic fluid and carcasses 11-day-old chick embryos infected with a strain of "seaside" virus of bird flu.

Use of antigenic material with biological activity is not lower than an 8.0 lg

EID50/cm3.

Purified antigenic material inactivate β-propiolactone.

β-propiolactone used in the form of a 10%aqueous solution.

β-propiolactone contribute to antigenic material to a concentration of 0.075%.

Inactivation of antigenic material β-propiolactone lead during 23-25 h at 24°C and pH 7.4 and 7.6.

Of the adjuvants used oil adjuvant Montanide ISA-70.

Oil adjuvant Montanide ISA-70 connect with antigenic material for emulsification in the amount of 60-70 wt.%.

<> The task in the part of the vaccine is solved by the fact that the vaccine inactivated emulsified against bird flu, containing the active substance and an oil adjuvant, according to the invention, as the active substance contains purified and avirulent antigenic material from strain "seaside", obtained as described above in an effective amount.

The vaccine contains purified and avirulent antigenic material from strain "seaside" and oil adjuvant at a ratio (wt.%):

Antigenic material30,0-40,0
Oil adjuvant60,0-70,0

The vaccine contains antigenic material with biological activity is not lower than an 8.0 lg EID50/cm3.

The vaccine of adjuvants include oil adjuvant Montanide ISA-70.

The original virus to obtain strain "seaside" is selected employees of the research Institute of Virology. Dijanoveckog in 2001 in the Primorsky region from wild ducks, who had no clinical signs of disease. Production strain "seaside" virus of bird flu with the antigenic formula H5N2obtained by multiple serial passages in chicken embryos. The strain Primorsk is th" deposited in the Public collections of microorganisms of the Federal state institution "all-Russian state Center for quality and standartizacii veterinary drugs and feed (FGI "VGNKI") under registration No. 129-DEPT.

The strain is non-pathogenic and is highly antigenic and immunogenic activity after inactivation. Experimentally confirmed the possibility of its use for the manufacture of inactivated vaccine.

Strain "seaside" virus GP is characterized by the following characteristics and properties.

Morphological properties

Strain "seaside" virus GP belongs to the family Orthomyxoviridae, genus Influenzae virus avicum, serotype a, subtype H5N2, has morphological features characteristic of orthomyxoviruses.

The virion has a spherical shape and a constant deposition 737 S. Virions polymorphic 80-120 nm is represented by two antigenic complexes. Outer V - antigen consists of hemagglutinin (H) and enzyme neuraminidase (N). The virion has a shell with small spines length of 10-12 nm, a diameter of 3-4 nm, which encompasses rolled in the ring nucleocapsid diameter of 8-9 nm. The chemical composition of the virion: the lipid - 23,5-25%, protein - 63,2%, carbohydrates - 7%, RNA and 1.8%. In the process of reproduction of the virus is the accumulation of the three types of particles: 1) Mature, complete with all the intrinsic properties of the virus; 2) "incomplete form", with only hemagglutinine activity; 3) in the form of soluble S-antigen, which has only complimentative activity.

Antigenic properties

Its ant the genetic properties of the strain "seaside" is identified as the virus of avian influenza type a, with the antigenic formula H5N2. The virus is stable neutralized homologous anticorodal.

The virus has an infectious, hemagglutinins, neuraminidase activity, agglutinate erythrocytes chickens, turkeys, Guinea fowls, pigeons, Guinea pigs, rats, mice, dogs, cows, pigs, cats, horses, and humans. When vaccination viral antigen induces the formation of specific antibodies detected in rtga in a dilution of 1:64-1:128.

Biotechnological characteristics

Strain "seaside" has a high biological activity in the native form, and demonstrate a high antigenic and immunogenic properties after inactivation. Strain "seaside" intended for use as raw material for the manufacture of inactivated vaccines and diagnostic Biologicals. Strain "seaside" reproducerea 11-day-old chick embryos. Within 48-72 hours of incubation the virus accumulates in extraembryonal fluid and the carcasses of chicken embryos in the title of 8.0 to 9.2 lg

EID50/ml. Strain "seaside" is stable.

Resistance to external factors

Hemagglutinin activity of influenza virus strain "seaside" is rapidly inactivated nitrous acid, but remains for a long time under the action of formalin, ether, trypsin, does not disappear when heated viral suspension to 56-60 is°. Exposure to the virus within 60 min temperature 60C°, ultraviolet rays, 1% sodium hydroxide solution, 2% solution of formaldehyde, nitrous acid, ether, chloroform completely inactivate it. It is rapidly inactivated at pH below 4.0 and above of 12.7. The virus reliably decontaminated with 1% sulfuric acid solution, 3% suspension of calcium hypochlorite, 2% sodium hydroxide solution, 5 % suspension of creoline, 5% phenol solution, 0.1% solution of mercuric chloride for 10-30 minutes In 50% glycerol strain "seaside" saves infectious properties for more than 5 months. Strain "seaside" does not lose infectious properties during freeze-drying.

Additional characteristics and properties

Immunogenic activity of 100%.

Pathogenicity is not defined.

Virulence is not defined.

Contagiousness is not defined.

Carcinogenesis is missing.

Based on the obtained data it can be argued that a strain of "seaside" antigenic and immunological spectra is original.

The invention is illustrated by examples of its execution.

Example 1

To obtain the vaccine from strain "seaside" matronym virus infect 11 - day chicken embryos derived from successful in infectious diseases farms. Virus-infected chicken embryos cultured for 72 hours at a temperature of up 37.0±0.5°. After 72 hours inkberrow the of infected chicken embryos are placed in the refrigerating chamber at a temperature of from 2 to 8C° and incubated for 16-24 hours. Chilled embryos under sterile conditions open, select extraembryonal liquid and carcass. Carcasses of infected chick embryos subjected to grinding in a colloid mill for 10-12 minutes, then vaccinated this mass is mixed with phosphate buffer solution, resulting in the 10-15%vaccinated suspension. Vaccinated suspension clear of ballast impurities in a known manner. The purified suspension is served in a sterile reactor and mixed with extraembryonal liquid. For virus inactivation using β-propiolactone. Pre-prepared 10%solution of β-propiolactone demineralised water. Then heated to 24°C vaccinated suspension make a 10%solution of β-propiolactone to a concentration of 0.075%. The mixture was mixed thoroughly and incubated at 24°C for 23-25 hours. After inactivation of the antigenic material is cooled to 2-8°C. To the cooled antigen with constant stirring add oil adjuvant as oil adjuvant use Montanide ISA-70 company "seppic" (France) or its analogs. Oil adjuvant for vaccine manufacturing sterilized according to the instruction supplied by the manufacturer.

Oil adjuvant and vaccine antigen is mixed on the homogenizer for 5 min at a rotation speed of the screw 4000 on the/min and a temperature of 10-15°C. You get a homogeneous white emulsion of the type water-in-oil".

Tested antigenic activity of a vaccine against avian influenza inactivated emulsified manufactured according to the following recipe

wt.%:
Cleaned and avirulent antigenic material from strain "seaside" virus GP type And subtype H5N2, received on 11 - day-old embryos SPF chickens with infectious activity 8,55 lg EID50/cm3 and HA-activity 1:25630,0
Oil adjuvant Montanide ISA-7070,0

In the experience used SPF chickens at the age of 25 days of 12 goal. Randomly selected 2 SPF chicken served as the control group, the remaining 10 SPF chickens were experienced by the group. SPF chickens from the experimental group by intramuscular injection in the breast area was administered the vaccine at a dose of 0.5 cm3.

After 21 days. after vaccination of SPF chickens experimental and control groups were taken blood samples, obtained blood serum, which according to the standard technique explored in rtga for antibodies to the virus of bird flu.

For setting rtha as positive control used the serum to the irus of avian influenza subtype H5N3 produced by OJSC Pokrov plant of biological products" (goknow, Russia), as negative control - normative serum of chickens (OJSC Pokrov plant of Biologicals", goknow, Russia), and the antigen is an inactivated virus of avian influenza subtype H5N2 strain "seaside". The number of tested sera corresponds to the number of chickens in the experience.

The results are shown in table 1.

Presented in table 1 the results of a study of sera in rtga indicate that the level of antibodies to the virus of avian influenza of subtype H5N2 through 21 days after immunization vaccine made 6.13 log2the proportion of Chicks that have a protective antibody titer (1:16) to the virus of avian influenza was 100%.

Example 2

Tested antigenic activity of a vaccine against avian influenza inactivated emulsified manufactured according to the following recipe

wt.%:
Cleaned and avirulent antigenic material from strain "seaside" virus GP type And subtype H5N2, received on 11-day-old embryos SPF chickens with infectious activity of 8.25 lg EID50/cm3 and HA-activity 1:12835,0
Oil adjuvant Montanide ISA-7065,0

p> In the experience used SPF chickens at the age of 23 days in the amount of 12 animals. Randomly selected 2 SPF chicken served as the control group, the remaining 10 SPF chickens were experienced by the group. SPF chickens from the experimental group by intramuscular injection in the breast area was administered the vaccine at a dose of 0.5 cm3.

After 21 days after vaccination from SPF chickens experimental and control groups were taken blood samples, obtained blood serum, which according to the standard technique explored in rtga for antibodies to the virus of bird flu.

For setting RTTA as positive control used the serum to the virus of avian influenza subtype H5N3 produced by OJSC Pokrov plant of biological products" (goknow, Russia), as negative control - normative serum of chickens (OJSC Pokrov plant of Biologicals", goknow, Russia), and the antigen is an inactivated virus of avian influenza subtype H5N2 strain "seaside". The number of tested sera corresponds to the number of chickens in the experience.

The results are shown in table 2.

Presented in table 2 the results of a study of sera in rtga indicate that the level of antibodies to the virus of avian influenza of subtype H5N2 through 21 days after vaccination was 6.4 log2the proportion of Chicks, imediately antibody titer (1:16) to the virus of avian influenza was 100%.

Example 3

Tested antigenic activity of a vaccine against bird flu emulsified inactivated manufactured according to the following recipe

wt.%:
Cleaned and avirulent antigenic material from strain "seaside" virus of avian influenza type a, subtype H5N2, obtained in 11-day-old embryos SPF chickens with infectious activity 8,45 lg EID50/cm3 and HA-activity 1:25640,0
Oil adjuvant Montanide ISA-7060,0

In the experience used SPF chickens at the age of 28 days of 12 goal. Randomly selected 2 SPF chicken served as the control group, the remaining 10 SPF chickens were experienced by the group. SPF chickens from the experimental group by intramuscular injection in the breast area was administered the vaccine at a dose of 0.5 cm3.

After 21 days after vaccination from SPF chickens experimental and control groups were taken blood samples, obtained blood serum, which according to the standard technique explored in rtga for antibodies to the virus of bird flu.

For setting rtha as positive control used the serum to the virus of avian influenza subtype H5N3 production of JSC "Pok the native plant biologics" (goknow, Russia), as negative control - normative serum of chickens (OJSC Pokrov plant of Biologicals", goknow, Russia), and the antigen is an inactivated virus of avian influenza subtype H5N2 strain "seaside". The number of tested sera corresponds to the number of chickens in the experience.

The results are shown in table 3.

Presented in table 3 the results of a study of sera in rtga indicate that the level of antibodies to the virus of avian influenza of subtype H5N2 through 21 days after vaccination was 6.73 x log2the proportion of Chicks that have a protective antibody titer (1:16) to the virus of avian influenza was 100%.

Example 4

Control of inactivated emulsified vaccine against bird flu from strain "seaside" with the antigenic formula H5N2 was carried out by determination of sterility, completeness of inactivation, harmlessness, antigenic activity of the drug. In addition, the vaccine was tested on the stability of the emulsion. Check for sterility performed in accordance with GOST 28085-89 "biological Preparations. Methods of bacteriological control of sterility". The method consists in determining the absence of growth of bacterial and fungal microflora in crops samples of the vaccine or antigen on nutrient media. To test for sterility is Rob vaccine or antigen shake and carry out the planting. In crops should not be the growth of bacterial and fungal microflora. The resulting vaccine was sterile.

Complete inactivation of the virus of avian influenza was determined by injection of ten chicken embryos 11-day age of the average sample of the vaccine in the amount of 0.2 cm3in allantoin cavity. The control were 5 non-infected chicken embryo. Incubated the embryos within 96 hours at t +37,2°C. the Destruction of embryos after 24 hours was considered nonspecific. After incubation chicken embryos were cooled at t° C 2 to 8°C for 24 h and were collected aseptically extraembryonal liquid, individually from each embryo. Then extraembryonal liquid explored in the DSA. If the result is negative, then spent the second passage, using for infection mixture extraembryonal fluid taken in equal amounts from each embryo. DSA with extraembryonal fluid obtained from chick embryos after 2 consecutive passages, was negative. In the manufactured vaccine there is no active virus.

Example 5

Tested the safety of a vaccine against bird flu emulsified inactivated. In the experience used SPF chickens at the age of 26 days in the amount of 36 goal., which were divided into three groups, 12 the goal. in each one. In each of the three groups randomly GRT the early 2 SPF chicken served as control subgroup, the remaining 10 SPF chickens were experienced by subgroup. The average sample vaccine each series in a volume of 1.0 cm3subcutaneously two injections of 0.5 cm3entered 10 SPF chickens corresponding to the experimental subgroups. Birds were monitored for 21 days, after which they were killed. The vaccine is considered harmless, if at necropsy of carcasses SPF chickens experienced subgroups on the vaccine has not been pronounced inflammatory response.

The results are shown in table 4.

Table 4 results of evaluation of the safety of inactivated emulsified vaccine against bird flu from strain "seaside"
VaccineHarmlessness
Sample 1Harmless
Sample 2Harmless
Sample 3Harmless

When viewed from the introduction of inactivated emulsified vaccine against bird flu from strain "seaside" with the antigenic formula H5N2 local lesions were not identified, suggesting the safety of this vaccine for poultry with the introduction of volume, 2 times pravilnoy about JEM.

Example 6

Tests of the physical properties of a vaccine against bird flu emulsified inactivated. The relative viscosity of the vaccine was determined on the viscometer VPI-2 and expressed in mm2/s2.

The emulsion stability was determined by centrifuging a sample of a vaccine. Three vials of vaccine field of intense shaking was selected by 12 cm3emulsion and carried it in three glass centrifuge tubes. Tubes emulsion was centrifuged at 3000 rpm for 30 minutes and then measured with a ruler the height of the column of clear fractions in the upper part of the tube.

The emulsion was considered stable if after centrifugation in each of the three tubes in the process of visual inspection did not find any content changes, or if the height of the column of clear fractions formed in the upper part of the tube, does not exceed 10 mm Not acceptable appearance after centrifugation clear fractions at the bottom of the tube or complete separation of the emulsion, when the drug loses its white color, decomposing into components: top oil and lower water fraction with the visible interface of two phases. The results are shown in table 5.

Table 5. pokazateli relative viscosity and stability of the emulsion inactivated emulsified vaccine against SE from strain "seaside"
VaccineViscosity,mm2/s2Centrifugation
Sample 147,5And 2% 98%
Sample 2to 43.1And 2% 98%
Sample 346,4A3% 97%

Note: fractions of emulsion: A - transparent oil liquid emulsion. With emulsion D - dense emulsion, E - water phase.

The results of testing the vaccine against avian influenza inactivated emulsified (series No. 1, 2, 3) on the physical properties indicate that the vaccine has a high emulsion stability and low relative viscosity from 43,1 to 47.5 mm2/s2.

Thus, the vaccine against avian influenza inactivated emulsified made on the basis of strain, "seaside", has a high antigenic activity and is able to ensure effective protection of susceptible birds from epizootic virus subtype H5.

Additional technical result from the use of the proposed method is due to the fact that inactivation of antigenic material is carried out by β-propiolactone that the call is authorized to significantly reduce labor and energy costs for the production of vaccines and to improve the quality of antigenic material.

1. The method of manufacture of inactivated emulsified vaccine against bird flu, including the preparation of antigenic material from a strain of the influenza birds, purification of antigenic material from the ballast impurities, the inactivation of the virus, the connection antigenic material with an oil adjuvant and control of the target product, characterized in that as the strain of the influenza birds use the strain "seaside" influenza of birds belonging to the family Orthomyxoviridae, genus Influenzae virus avicum, serotype a, subtype H5N2, a collection of FGI VGNKI, registration No. 129-DEPT.

2. The method according to claim 1, characterized in that to obtain the antigenic material used carcass 14-day-old chick embryos infected with the pathogen.

3. The method according to any one of claims 1 and 2, characterized in that the antigenic material used in the form vaccinated suspension, made of extraembryonic fluid and carcasses 14-day-old chick embryos infected with a strain of "seaside" virus of bird flu.

4. The method according to claim 3, characterized in that the use of antigenic material with the biological activity of not less than 8,0lg EID50/cm3.

5. The method according to claim 1, characterized in that the purified antigenic material inactivate β-propiolactone.

6. The method according to claim 5, characterized in that the β-propiolactone used in the form of 10%in the aqueous solution.

7. The method according to any of pp.5 and 6, characterized in that the β-propiolactone contribute to antigenic material to a concentration of 0.075%.

8. The method according to claim 7, characterized in that the inactivation of antigenic material β-propiolactone lead during 23-25 h at 24°C and pH 7.4 and 7.6.

9. The method according to claim 1, characterized in that the adjuvant use of an oil adjuvant Montanide ISA-70.

10. The method according to claim 9, characterized in that the oil adjuvant Montanide ISA-70 connect with antigenic material for emulsification in the amount of 60-70 wt.%.

11. Vaccine inactivated emulsified against bird flu, containing the active substance and an oil adjuvant, characterized in that the active substance contains purified and avirulent antigenic material from strain "seaside", obtained according to any one of claims 1 to 10, in an effective amount.

12. The vaccine according to claim 11, characterized in that it contains purified and avirulent antigenic material from strain "seaside" and oil adjuvant at the ratio, wt.%:

Antigenic material30,0-40,0
Oil adjuvant60,0-70,0

13. The vaccine according to claim 11, characterized in that it contains antigenic material with the biological activity of not less than 8,0lg EID50 /cm3.

14. The vaccine according to claim 11, characterized in that adjuvants include oil adjuvant Montanide ISA-70.



 

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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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