Strain 02-ag ru 09ru3124 of human immunodeficiency virus type 1, recombinant subtype 02-ag used for diagnosing and studying efficacy of therapeutic and vaccine preparations

FIELD: medicine.

SUBSTANCE: strain is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare, No. V-416.

EFFECT: strain can be used for developing and improving HIV diagnostic techniques, studying the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, and for creating a national panel of HIV-1 strains.

3 dwg, 3 ex

 

The invention relates to a strain of human immunodeficiency virus first type belonging to the recombinant subtype 02_AG, and can be used in Virology, medicine and biotechnology. Strain is a convenient natural model and can be used to develop and improve methods of diagnosis of HIV infection, study of the effectiveness of therapeutic and preventive chemotherapy and vaccines, as well as to create a national panel of strains of HIV-1.

A high degree of genetic heterogeneity of the human immunodeficiency virus is a major challenge faced by researchers. The variability of the nucleotide structure of the virus contributes to the rapid emergence of isolates resistant to neutralizing antibodies, cytotoxic T lymphocytes and antiviral drugs. For the development of various vaccines need to take into account the high variability of the viral genome. Currently in Russia there is no representative of the Bank is characterized strains of HIV-1. Therefore, we need to work on the selection, molecular biological characterization of isolates of HIV-1 and create a collection of HIV-1, which is necessary to study the properties of the new vaccine and therapeutic drugs. This collection should include epidemiologically significant Varian is s virus, reflecting the heterogeneity of HIV-1, the most characteristic of the individual regions or at-risk groups (drug users, gay men, nosocomial infections in the hearth, and so on).

In the development of the epidemic process of HIV spread in the Russian Federation plays a Central role subtype a HIV-1. However, in 2008, registered a sharp increase in the proportion of recombinant variants 02_AG circulating among genetic variants of HIV-1. The rapid growth in the spread of new genetic variants of HIV-1 in Russia gives greater urgency to obtain and study of strains of this subspecies, as domestic isolates and strains of subtype 02_AG not described in literature.

A known strain hominis immunodeficit virus HIV-1/Russia GM-12-95 (EN-1295) subtype In subgroups lentiviridae family Retroviridae for the preparation of diagnostic and vaccine preparations (patent RF №2121502, IPC C12N7/00, publ. 10.11.1998)containing the following amino acid sequence region V3: CTRPNNNTRKRIHIGPGRAYFTGRIIGDIRQAYC.

However, because most widespread in the Russian Federation acquiring the human immunodeficiency viruses type 1, belonging to subtypes a and 02_AG, which found the majority of people infected with this strain is not suitable for diagnostic purposes and to examine the effectiveness of the treatment routine is fir and vaccines, used on the territory of Russia.

Known isolates of human immunodeficiency virus type 1, including a group of isolates belonging to the recombinant subtype 02_AG obtained in Thailand (Development of a Panel of Well-Characterized Human Immunodeficiency Virus Type 1 Isolates from Newly Diagnosed Patients Including Acute and Recent Infections//AIDS RESEARCH AND HUMAN RETROVIRUSES. Volume 25, Number 1, 2009), which are used in the panel for the study of antiviral drugs.

However, these isolates were not suitable for diagnostic purposes and to study the effectiveness of therapeutic and preventive vaccines used on the territory of the Russian Federation, as they differ in genetic characteristics of isolates 02_AG HIV-1 spread in Russia.

The closest analogue (prototype) are isolates of human immunodeficiency virus type 1, including a group of isolates belonging to the recombinant subtype 02_AG obtained also in Thailand, the same group of researchers (Biologic and Genetic Characterization of a Panel of 60 Human Immunodeficiency Virus Type 1 Isolates, Reprezenting Clades A, B, C, D, CRF02_AG, for the Development and assessment of Candidate Vaccines//Jornal of virology, Mai 2005, p.6089-6101), which are used in the panel for the study of antiviral drugs.

However, these isolates due to genetic differences are not suitable for diagnostic purposes and to study the effectiveness of therapeutic and preventive vaccines what's drugs, used on the territory of the Russian Federation.

The technical result of the invention is to obtain such a strain of HIV-1, which would be suitable for diagnostic purposes and to study the effectiveness of therapeutic and preventive vaccines used on the territory of the Russian Federation.

This technical result is achieved by obtaining a new strain 02_AG.RU.09RU3124 of human immunodeficiency virus type 1 recombinant subtype 02_AG, which is allocated on the territory of Russia from the patient 28 years old, infected with sexually transmitted, diagnosed with acute HIV infection. The strain intended for inclusion in the national panel of strains, and can also be used to study the effectiveness of therapeutic and preventive chemotherapy and vaccines, improved methods of diagnosis of HIV infection.

The inventive strain of human immunodeficiency virus HIV-1 02_AG.RU.09RU3124 deposited in the Collection of microorganisms fsri SRC VB "Vector" of Rospotrebnadzor number V-416 from 15.06.09.

Figure 1 shows the nucleotide sequence of the pol gene of HIV-1 strain 02_AG.RU.09RU3124. Figure 2 shows the dynamics of accumulation virusspecific protein P24 in the mixed culture of the IPC, infected with strain 02_AG.RU.09RU3124, and figure 3 - dynamics of accumulation virusspecific protein P24 in infic the level of strain 02_AG.RU.09RU3124 culture cells MT-4 and U-937.

Description of the proposed strain

The new strain has a number of virological characteristics, which allows to consider it as a convenient natural model for evaluating the effectiveness of new tools for the diagnosis and research of new therapeutic and preventive drugs. The strain has a high reproductive activity, the type of replication refers to the category of Rapid/high, easily propagated in neoplastic T-lymphocyte MT-4 and monocytic cells U-937. About the products of the virus can be easily judged by a pronounced cytopathic effect and accumulation virusspecific protein P24. The strains stored at - 70°C.

Identification of HIV-1 strain 02_AG.RU.09RU3124 implemented method of determining the nucleotide sequences of fragments of the genome of HIV-1 coding region of the gene pol and the main envelope protein (env gene), automated sequencing machine (ABI PRISM. Subsequent phylogenetic analysis and identification of the subtype of the virus strain was performed using the program presented on the website of Stanford University http://hivdb.stanford.edu). Determination of tropism to isolate the cellular chemokine used was performed using available on the web sites of the programs Geno2pheno (http://coreceptor.bioinf.mpi-inf.mpg.de and WebPSSM and (http://indra.mullins.microbiol.washington.edu/webpssm).

Cultural properties. The virus strain isolated from limpii the s peripheral blood of a patient with a diagnosis of Acute HIV infection. Upon receipt of the isolate used method cocultivation lymphocytes infected patient blood lymphocytes of a healthy donor, stimulated by the mitogen, and the method of infection of human neoplastic suspension transplantable cell lines MT-4 and LI-937.

A reproduction. The strain of the virus replicates in peripheral blood lymphocytes, lymphoblastoid cell cultures MT-4 and U-937. Virus reproduction is accompanied by a characteristic cytopathic effect of mixed type - the death of individual cells, the formation of entities of 5-10 cells and accumulation virusspecific protein P24. The HIV-1 strain 02_AG.RU.09RU3124 can be maintained for a long time when passirovannye in a mixed culture of peripheral blood lymphocytes of healthy donors, in the culture of cells MT-4 or U-937. After 4-7 days after infection of the cells of the vaccinated culture fluid of the infected cell suspension is centrifuged at 1500 rpm and frozen in solution for cryopreservation (70% bovine serum, 10% glycerol) at - 70°C. After thawing is brought in fresh cell suspension at a ratio of 1:10.

Reproductive characteristics of the isolate is determined by the method of enzyme-linked immunosorbent assay for the accumulation virusspecific protein P24 in the model above cell cultures(figure 2, 3).

The infectivity of the viral strain was assessed by the accumulation virusspecific protein P24 in the culture fluid of 7 and 15 days of cultivation mixed culture stimulated phytohemagglutinin (PHA) of the IPC from two seronegative donors after infection of these cells growth medium, taken at 7 and 15 days of cultivation allocated isolate by making the growth medium into the culture of the IPC (1:10).

Antigenic properties. HIV-1 belongs to the family retroviridae podgroup lentiviridae. Viruses of this subgroup are enveloped RNA viruses with a particle size of about 100-150 nm. In the composition of the particles find no less than 6 major structural antigens having immunogenic properties that can be detected in infected cells using various immunological and virological methods.

Analysis of the nucleotide sequence of the genome, phylogenetic analysis and identification of the subtype of the virus strain was performed using the program presented on the website of Stanford University http://hivdb.stanford.edu). Genotyping of the obtained strain of HIV-1 02_AG.RU.09RU3124 conducted by analyzing the nucleotide sequences of the pol gene. The choice of this genetic area was determined by the fact that their analysis allows to distinguish the main variants of HIV-1 circulating in the Ter is itoria of the former USSR. For the definition and analysis of the nucleotide sequence of the pol gene was used sets "Viroseq" ("Viroseq HIV-1 Genotyping System") of the company "Celera Diagnostics" (USA), and "AmpliSens HIV genotype" company "Interlabservice (Russia). Determination of tropism to isolate the cellular chemokine used was performed using available on the web sites of the programs Geno2pheno (http://coreceptor.bioinf.mpi-inf.mpg.de and WebPSSM (http://indra.mullins.microbiol.washington.edu/webpssm).

Example 1. Genotyping of HIV-1 strain 02_AG.RU.09RU3124. Genotyping of the obtained strain of HIV-1 02_AG.RU.09RU3124 conducted by analyzing the nucleotide sequences of plots genes pol and env. Data selection genetic plots was determined by the fact that their analysis allows to distinguish the main variants of HIV-1 circulating in the territory of the former USSR. For the definition and analysis of the nucleotide sequence of the gene ro use sets "Viroseq" ("Viroseq HIV-1 Genotyping System") of the company "Celera Diagnostics" (USA), and "AmpliSens HIV genotype" company "Interlabservice (Russia).

Virusspecific RNA extracted from serum samples. Virusspecific DNA obtained by the method of reverse transcription of viral RNA and used for subsequent amplification using nested polymerase chain reaction. Determination of the nucleotide sequences carried out automatically on the device model h firm "Aplaud Biosystems (USA).

The obtained nucleotide sequence that is midna sequence for gene RA genome of HIV-1 include a full nucleotide sequence of the gene of the protease and partial sequence of a gene reverse transcriptase - 1323 nucleotide residues (figure 1).

The obtained data show that HIV-1 isolate 02_AG.RU.09RU3124 is the original strain of HIV-1 and belongs to the subtype 02_AG. Conducted using Geno2pheno and WebPSSM analysis indicates that HIV-1 isolate 02_AG.RU.09RU3124 has the greatest trapnest to the cellular chemokine used CCRS-type.

Conducted using jpHMM-HIV (http://jphmm.gobics.de the analysis showed that the isolate 02_AG.RU.09RU3124 characterized by unique genetic organization of the locus pol. This locus has mosaic recombinant structure - from 2253 BP to 3106 P.N. (numbering according to reference strain NGV-2), homologous subtype G, and from 3168 P.N. up 3575 gel - subtype A1, and localization of the site of recombination in the field 3106-3168 P.N.

Example 2. For selection of primary isolates of HIV-1 in peripheral blood mononuclear cells (IPC) from the patient is cultured with a mixed culture stimulated phytohemagglutinin (PHA) of the IPC from two seronegative donors. The selection of the IPC carried out by centrifugation of whole blood on a Ficoll gradient. PHA stimulation donor IPC spend 2-3 days (5 μg PHA/ml). For cocultivation mix 4-6×106IPC cells of a patient with 8-12×106IPC donor cells in 10 ml growth medium (PC) of the following composition is RPMI-1640, 20% fetal bovine serum (red), 100 e.a./ml IL-2, 300 mg/ml glutamine, 150 μg/ml link is Mitina and 100 μg/ml of gentamicin. Every 3-4 days carry out the replacement of the culture medium. Selected culture fluid examined for the content of P24 antigen and are cryopreserved at - 70°C. For 7 and 15 day cocultivation after replacing the culture medium to the remaining infected cells add 4-6×106PHA-activated donor of the IPC. Virus replication was assessed by the accumulation of virusspecific protein P24. Virusspecific protein P24 detected in the culture fluid by the method of enzyme-linked immunosorbent assay using enzyme immunoassay system for the detection and confirmation of the presence of P24 antigen of HIV-1 "Vuckovic-1 P24-antigen-confirming test Fund 42-0117-5686-04", with a sensitivity of 5 PG/ml for CCA risk., or set of reagents for immunoassay to detect antibodies to HIV-1,2 and antigen P24 HIV-1 "Cambiaste-1.2 AG/at, THE 93 98-098-23 548172-2007".

The figure 2 presents the results of the accumulation of P24 in the culture fluid in the culture of HIV-1 strain in a mixed, activated mitogen culture IPC, reflecting the dynamics of the reproduction of the virus. The strain is characterized by extremely high rate of viral protein P24 and the protein concentration is already on the 11th day reaches the maximum value (208500 PCG/ml). The type of replication this strain can be classified as Rapid/high.

Example 3. The study on which opline virus in various cell cultures. The suspension of cells MT-4 or U-937 infect the culture fluid from an infected mixed culture IPC method joint incubation in the ratio of 5×106cells in 1 ml of culture fluid for 1 h at 37°C. Centrifuged at 1000 rpm for 5 min, remove supernatant, cells resuspended in a nutrient medium RPMI-1640 with the addition of 10% fetal serum, previously inactivated by heating at 56°C for 30 minutes, 300 mg/ml L-glutamine, 80 μg/ml gentamicin and 30 mg/ml of lincomycin to a concentration of 0.5×106cells in a milliliter. Vials with cell culture incubated in a thermostat at 37°C. Control of the cytopathic effect of the virus and sensitivearea carried out by the method of inverted microscopy of cell suspension in the culture bottles. Every 3-4 days carry out the replacement of the culture medium. Selected culture fluid examined for the content of P24 antigen and are cryopreserved at -70°C. On day 10 of cultivation after replacing the culture medium to the remaining infected cells add a suspension of uninfected cell cultures in growth medium with a concentration of 1×106cells/ml of virus Replication was assessed by the accumulation of virusspecific protein P24. Virusspecific protein P24 detected in the culture fluid by the method of tverdofaznogo the enzyme immunoassay using enzyme immunoassay system for the detection and confirmation of the presence of P24 antigen of HIV-1 "Vuckovic-1 P24-antigen-confirming test Fund 42-0117-5686-04", with a sensitivity of 5 PG/ml for CCA risk., or set of reagents for immunoassay to detect antibodies to HIV-1,2 and antigen P24 HIV-1 "Combivent HIV-1,2 AH/at, THE 9398-098-23548172-2007".

The figure 3 presents the results of the accumulation of P24 in the culture fluid in the culture of the isolate of HIV-1 in neoplastic T-lymphocytic cells MT-4 and monocytic U-937, reflecting the dynamics of the reproduction of the virus. The strain is characterized by different speed and dynamics of the accumulation of viral protein P24 in different cell cultures. The maximum concentration in macrophage culture U-937 is achieved on the 17th day of cultivation, and neoplastic T-lymphocytic cells MT-4 - on day 10. The observed concentration of the protein P24 in culture isolate on macrophage culture U-937, compared with cultivation on neoplastic T-lymphocytic cells MT-4, probably due to the fact that this isolate has the highest trapnest to the cellular chemokine used CCR5-type. The chemokine can be used CCR5-type, as used CXCR4-type provided on the surface of cells U-937, whereas on the surface of cells MT-4 are only used CXCR4-type.

The inventive strain of HIV-1 02_AG.RU.09RU3124 used in the development of a national panel of genetically and biologically characterized strains of HIV, have epidemiological significance on the territory of Russia for the standardization of the assessment of the effectiveness of the developed vaccine and antiviral drugs. Panel to assess the effectiveness of the vaccine is created from strains isolated from patients in the acute stage of HIV infection. In addition, the panel should include genetically and biologically heterogeneous strains belonging to the same subtype of HIV-1, representing the maximum diversity possible circulating on the territory of viral variants that would allow a more adequate and correct evaluation of the antiviral effects of drugs on the model as close as possible to the real processes occurring in the body of an infected HIV-1 person.

The inventive strain 02_AG.RU.09RU3124 HIV-1 is supposed to be used also in the panel to assess the effectiveness of antiviral drugs. Panel to assess the effectiveness of antiviral drugs includes genetically and biologically different HIV strains isolated from HIV-1-infected patients at different stages of the disease and including patients taking antiviral drugs. The presence of such panels in Russia will allow standardized adequate assessment of promising antivirals for treatment of HIV infection, what is the actual problem for Ross and, because our country has the scientific potential for drug development, but not certified tool for screening drugs for their effectiveness against HIV-1 on the model of the live virus.

In addition, the claimed strain of HIV-1 can be used as a producer of antigens which are diagnostic kits and test systems, in particular antigens of HIV-1, required for the production of immunoassay diagnostic test kits for detection of HIV infection and kits for immunoblot confirming HIV infection in newly diagnosed patients.

Strain 02_AG.RU.09RU3124 of human immunodeficiency virus type 1 recombinant subtype 02_AG used for the diagnosis and study of the efficiency of therapeutic and preventive vaccines, deposited in the Collection of microorganisms fsri SRC VB "Vector" of Rospotrebnadzor number V-416 and having the nucleotide sequence of the gene l shown in figure 1.



 

Same patents:

FIELD: medicine.

SUBSTANCE: presented strain of human immunodeficiency virus HIV-1 A1.RU.09RU2225 is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare, No. V-391.

EFFECT: strain can be used for developing and improving HIV diagnostic techniques, studying the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, and for creating a national panel of HIV-1 strains.

4 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: presented strain of human immunodeficiency virus HIV-1 A1.RU.09RU2065 is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare, No. V-389.

EFFECT: strain can be used for developing and improving HIV diagnostic techniques, studying the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, and for creating a national panel of HIV-1 strains.

4 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: method of detecting anthrax pathogen in environment and biological fluids is based on functional detection of activity of one of anthrax toxin components, weak protease of anthrax lethal factor (LF). Applied principle of highly sensitive detection of LF proteolytic activity is based on the system of amplification of sugnal, originating from the act of substrate cutting, by means of present in substrate composition auxiliary enzyme of alkaline phosphatase of Escherichia coli (AP). In composition of recombinant LF substrate in addition to alkaline phosphotase and specific for LF substrate sequence RRKKVYPYPME, there is a peptide which is biotinilated in vivo and in vitro by means of biotin-ligase of E.coli and ensuring substrate immobilisation on solid phase (plaque surface) due to interaction with avidin and its derivatives, and sequence of six histidines for purification of recombinant substrate from periplasm of E.coli by means of metal-chelate resin.

EFFECT: method in accordance with the invention possesses high sensitivity, with application of fluorescein diphosphate makes it possible to detect up to 1 pM of lethal factor, rapidity of execution, high specificity, low risk of false-positive signals.

1 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: assessment of the clinical efficacy of antibacterial preparations is three-staged; the first stage involves working up an antibiotic-sensitivity criteria scale for reference strains; at the second stage the antibiotic-sensitivity of an investigated material is evaluated, while the third stage consists in correlating the latter values with the derived scale for a given type of microbe; the first and second stages are simultaneous and based on the same technique by inoculation of a microbial suspension in the amount 0.3 ml (n·107CFU/ml) by a diffusion test and a dilution test on a nutrient medium with growth supplements. Thereafter, the inoculation is incubated at 37°C for 24-48 hours, and the results are analysed. The assessment of the clinical efficacy is enabled by correlating the border values of minimal inhibitory concentration and growth inhibition zone diametres with the relevant criteria scale. If the values of the investigated material fall within a permissible range of the criteria scale, an antibiotic is considered to be effective.

EFFECT: use of the method enables the reliable and immediate assessment of the clinical efficacy of antibacterial preparations for tularemic and brucellous infections.

7 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: substance of the invention involves the method for producing an erythrocytic antigen for indirect hemagglutination test (IHT), by preparing formalinised erythrocytes to be antigen-pulsed macrophage; after cultivation, the bacterial mass in amount 50-60 billion microbial cells in 1 ml is inactivated by processing by 2% concentrated secondary sodium alkyl sulfate detergent at 0.75-10 ml of the antigen in 1 ml of a 10% of an erythrocyte suspension and kept for 2 hours at 45°C.

EFFECT: improved specificity of a diagnosticum.

1 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: after provisional diagnosing of a vascular disease, patient's blood serum is analysed for beta2-glycoprotein 1 antibodies (B2GPab), endothelium antibodies (Eab), Clq complement factor antibodies (Clq ab), cryoglobulins (CG) in the form of cryocrit and activity of rheumatoid factor (RF). If B2GPab is equal to 5 standard unit/ml and more, Eab is equal to 1:10 and more, Clq ab is equal to 5 standard unit/ml and more, CG in the form of cryocrit is more than 0%, RF is equal to 1:20 and more, therapeutic intervention which involves four daily procedures of leucopheresis with klion autoleukocyte adsorption in a single dose 500 mg combined with oral administration of biseptol in a single dose 480 mg is executed. After 4-5 weeks from the beginning of therapy, control blood examination for infection agents with the use of culture and genetic diagnostic techniques follows. If observing bacterial agents, persistent latent bacterial infection with associating vessel wall involvement is diagnosed.

EFFECT: higher diagnostic accuracy of chronic latent bacterial infection with associating vessel wall involvement.

3 tbl, 3 ex

New virus of plants // 2411290

FIELD: agriculture.

SUBSTANCE: first plant or its part is exposed to infection dose of ToTV. Then plants with no or slight symptoms of disease are identified.

EFFECT: plants identified in such a manner as resistant to virus are used as donor ones to cross with recipient plants, and plants resistant to ToTV are chosen from descendant plants.

8 cl, 7 dwg, 5 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention can be used for obtaining diagnostic and/or therapeutic agents, in particular antibodies, which specifically interact with proteins of cell surface of target cells. Claimed is method of screening phage display library and including it method of isolation of library element, which is specific partner of binding of target cell surface protein, and method of isolation of unknown cell surface protein, specifically interacting with library element. Method of screening phage display library according to invention includes contact of said library with cells of interest with further separation of cells, which bound with one or several elements of expression library, from non-bound elements by separation through organic phase, and differs by introduction of additional stage, which consists in carrying out before said separation of at least one stage of material washing, and ensures essential increase of method efficiency.

EFFECT: higher efficiency.

30 cl, 7 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: essence of invention includes contact of liquid sample, taken from mammal organism, with one or several monoclonal antibodies to Lawsonia intracellularis antigen, secreted by cell lines of ECACC hybridomes, which have registration numbers. Invention also includes diagnostic test-kit, containing antibodies specific for Lawsonia intracellularis.

EFFECT: increased specificity of antigen detection.

11 cl, 5 ex, 5 dwg

FIELD: medicine.

SUBSTANCE: kit includes polyclonal antibodies against protein of said viral envelope and a pair of primers: upstream 5'-primer with ID 1: CTGCAGATGGTTTGCCGAATTTGCAA sequence and downstream 3'-primer with ID 2: GCTCTAGACTAGATCTCAAGCAGGTC sequence.

EFFECT: kit allows detecting Prunus necrotic ring spot virus in plants.

17 cl, 4 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: presented strain of human immunodeficiency virus HIV-1 A1.RU.09RU2225 is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare, No. V-391.

EFFECT: strain can be used for developing and improving HIV diagnostic techniques, studying the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, and for creating a national panel of HIV-1 strains.

4 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: presented strain of human immunodeficiency virus HIV-1 A1.RU.09RU2065 is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare, No. V-389.

EFFECT: strain can be used for developing and improving HIV diagnostic techniques, studying the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, and for creating a national panel of HIV-1 strains.

4 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: offered is a vaccine containing dimerethylenimine inactivated goose parvovirus antigen of Goosa Parvovirus Strain, VGNKI No.71, clone 6 of the activity not less 6.5-7.5 lg "ТЦД"50/cm3 mixed with the oil adjuvant Montanide ISA-70, taken in the ratio 1:1-1:4; the dimerethylenimine concentration is not less than 0.1%. A method of goose parvovirus infection vaccination includes the intramuscular introduction of the vaccine preparation to a bird of a parental flock in a single dose 2.5·104-2.5·105 "ТЦЦ"50/cm3 30 days prior to the expected oviposition.

EFFECT: vaccine preparation is harmless for the birds, exhibits the evident antigenic and immunogenic activity, invokes an intense long immune response in geese for the entire reproductive period; the single application of the inactivated preparation is more technological and saving, reduces immune system stresses and loads in the birds.

2 cl, 2 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: it is described that immunogenicity of hemagglutinin (HA) molecule of influenza virus can be increased by substituting amino acids in the HA sequence. Substituting specific residues in HA such as asparagine introduction in position 223 in HA H5 allows ensuring more sensitive hemagglutination inhibition (HI) test provided by changing receptor specificity and/or ability to antibody-antigen linkage. The HA molecules having such substitutions can find application in creating diagnostic prototype viruses.

EFFECT: improved influenza virus vaccines.

9 cl, 3 dwg, 7 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine and deals with method of obtaining lice culture influenza vaccine. Essence of the invention includes method of obtaining virus-containing substance by cultivation of one of cold-adapted influenza virus reassortants with enoculation dose, with multiplicity of infection not lower than 0.0001 EID50/cell in MDCK cell culture on micro-carriers, which have concentration not less than 1 g/l, with application as micro-carrier material of porous polypropelene, in supporting serum-free nutritional medium, containing proteolytic enzyme in amount 0.25-50.0 mcg/ml, and stabilising additive, which includes sorbitol, or sucrose, or peptone from soya in concentration 0.5-4.0 wt%, collection of virus-containing liquid after cultivation is carried out at least 2 times when specific influenza virus activity before each collection of virus-containing liquid reaches at least 7.0 Ig EID50/ml, concentration and purification of virus substance from ballast admixtures, introduction into purified substance before drying of stabilising additives, with application as such of either proline, glycene, lactose, glutamine-acidic sodium, sucrose, gelatins in final concentration (1.5-5), (1.5-5), (1.5-10), (1.5-5), (5-30) and (1-10) wt % respectively, or sucrose, gelatose and soya peptone in final concentration (1-8), (1-8) and (1-8) wt % respectively, or sorbitol and gelatose in final concentration (3-8) and (3-8) wt % respectively.

EFFECT: obtaining more thermostable vaccine with high output.

2 cl, 6 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: strain is deposited in the collection of VGNKI, registration name Belgorod-03 - DEP rabbit hemorrhagic disease virus. The strain is used for preparing vaccines and diagnostic products.

EFFECT: high infectious, antigenic and immunogenic activity.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: disclosed is a plasmid for producing a viral vector transporting multiple expression cassettes to a target. The plasmid contains genome nucleotide sequences packed into a polyvalent capsid and a number of cassettes to be transported. A method for producing a viral vector, a viral vector and an immunogenic composition are described besides.

EFFECT: group of inventions can be used for wide-ranging expression of target antigens.

23 cl, 3 dwg

FIELD: medicine.

SUBSTANCE: shown is antigen and immunogenic efficacy of a new Manichino-09 strain used for preparing an inactivated RHDV vaccine. The vaccine contains an antigen material of the Manichino-09 strain of rabbit hemorrhagic disease virus, formalin, sodium metabisulphite, aluminium hydroxide gel in the effective proportions. The strain is deposited in the collection of VGNKI, registration name Manichino-09 - DEP rabbit hemorrhagic disease virus. The virus is reproduced in vivo in a European rabbit (Oryctolagus cuniculus) of 1.5 month and older, free from the disease antibodies or having a RHDV antibody titre lower than 1:4. An inactivant is 4% formalin (i.e. 1.1-1.4% formaldehyde). The sorbed vaccine is prepared of adjuvants with using aluminium hydroxide gel (AHG).

EFFECT: vaccine exhibits high antigen and immunogenic activity.

3 tbl

FIELD: medicine.

SUBSTANCE: vaccine strain of A/17/Brisbane/07/28 (H1N1) influenza virus is a reassortant prepared by cross breeding an epidemic A/ Brisbane /59/07 (H1N1) virus and a cold-adapted thermally sensitive A/Leningrad/134/17/57 (H2N2) virus being a human-safe attenuation donor. The A/17/Brisbane/07/28 (H1N1) strain replicates actively in growing chicken embryos at optimum temperature 32°C. The reassortant has inherited the genes coding surface virus antigens - hemagglutinin (HA) and neuraminidase (NA) from an epidemic parent virus, and six genes coding internal non-glycosylated proteins from the attenuation donor. The A/17/Brisbane/07/28 (H1N1) strain is areactogenic for adults and children in intranasal introduction. In reactogenecity, the vaccine strain of A/17/Brisbane/07/28 (H1N1) influenza virus meets the requirements specified in the Manufacturer's Monograph 42-0417-4097-03 to vaccine strains for live dry allantoic influenza vaccine for intranasal application.

EFFECT: strain is characterised by thermal sensitivity and cold adaptation.

1 dwg, 3 tbl

FIELD: medicine.

SUBSTANCE: A/17/Brisbane/07/1 (H3N2) strain replicates actively in growing chicken embryos at optimum temperature 32°C. A reassortant has inherited the genes coding surface virus antigens - hemagglutinin (HA) and neuraminidase (NA) from an epidemic parent virus. The other six genes coding internal non-glycosylated proteins have been inherited from an attenuation donor. The A/17/Brisbane/07/1 (H3N2) strain is areactogenic for adults and children in intranasal introduction. The invention can be used in practical health services for preventing seasonal influenza incidence in adults and children by a live influenza intranasal vaccine of the A/17/Brisbane/07/1 (H3N2) strain.

EFFECT: strain is characterised by thermal sensitivity and cold adaptation.

1 dwg, 3 tbl

FIELD: medicine.

SUBSTANCE: presented strain of human immunodeficiency virus HIV-1 A1.RU.09RU2225 is deposited in the Collection of Microorganisms of Federal State Research Institution State Science Centre Vector of Federal Service for Supervision of Consumer Rights and Human Welfare, No. V-391.

EFFECT: strain can be used for developing and improving HIV diagnostic techniques, studying the efficacy of therapeutic and preventive chemotherapeutic and vaccine preparations, and for creating a national panel of HIV-1 strains.

4 dwg, 3 ex

Up!