Complex antigen of measles virus used as a component of immunoenzymometric test system for diagnostics of antibodies to measles virus
SUBSTANCE: complex antigen of measles virus used as a component of immunoenzymometric test system for diagnostics of antibodies to measles virus was produced from culture liquid containing measles virus stain Leningrad 16 with titer no less than 105.0 50% tissue cytopathic dose/ml, Edmonston - no less than 106.0 50% tissue cytopathic dose/ml, NovO/96 no less than 108.0 50% tissue cytopathic dose/ml by inactivation of its infectious activity by detergent and separation of protein from cellular lysate by chromatographic purification in amount no less than 2 mcg/ml with the purity no less than 70%. Culture virus-containing liquid was processed using separate cultivation of measles virus stains Leningrad 16, Edmonston and NovO/96 on monolayer of Vero cells culture with subsequent mixing of culture virus containing liquid in proportion 1:1:1 v/v.
EFFECT: Usage of invention provides for increased sensitivity and specificity of a complex antigen possessing the property to detect antibodies in blood serum.
2 cl, 4 tbl, 6 ex
The invention relates to a new complex antigen measles virus for use as a component of immunoassay diagnostic test systems and can be used in medical Virology and Microbiology.
Known antigen measles virus, derived from the strain of measles virus Leningrad 16 (L-16)is used as a component of a diagnostic test system (Guide to vaccine and serum case under the editorship of Academician of AMS Pinboliada. M: "Medicine", 1978. - s-223). Development of technology for antigen measles virus strain L 16 on the tissue culture of embryos of the Japanese quail (Vasiliev GA, Boychuk L.M. - "Specific prevention of measles". Materials of scientific-practical conference of VNIIEM them. Pasteur. - L., 1970. - s-210).
However, this antigen measles virus provides insufficient sensitivity and specificity of antibody detection using test systems based on it.
Known antigen measles virus, derived from the strain of measles virus, Edmonston (U.S. Patent No. 4211843, IPC A01N 1/02; AC 39/12, publ. 08.07.1980 year). Strain Edmonston deposited in the American Cell culture Collection (ATSS - VK-24, 4/92). In SRC VB "Vector" has been four passage on the culture of Vero cells. The authenticity of the installed strain by the method of positive polymerase chain reaction with specific pry the apostrophes. The strain belongs to genotype A. the Maximum titres reach 5-6 days after infection of a monolayer of Vero cells constitute 5×106TCPD50/ml [Agafonov A., et al.// Vopros., 1997, N.1, pp. 102-205]. Antigenic properties of strain: detects antibodies to measles virus in the sera of patients and vaccinated people.
However, this antigen measles virus provides insufficient sensitivity and specificity of antibody detection using test systems based on it.
The closest analogue (prototype) is the antigen measles virus, derived from the strain of measles virus NovO/96 and used as a component of an enzyme immunoassay system for the diagnosis of antibodies to measles virus (patent RF №2230785, IPC C12N 7/00, publ. 20.06.2004,). The virus develop on the monolayer of Vero cells, destroy cells by freeze-thawing, the lysate of the cells obtained by centrifugation and ultrafiltration. The antigen for ELISA clear of foreign proteins by ultracentrifugation in a density gradient of sucrose, which inactivate by heating at 56°C for 30 minutes
However, this antigen measles virus also has limited sensitivity and specificity of antibody detection using test systems based on it.
The technical result of the claimed invention is increased. is a major and specificity of complex antigen measles virus, providing detection of antibodies in serum.
This technical result is achieved by obtaining the complex of antigen measles virus used as a component of an enzyme immunoassay system for the diagnosis of antibodies to measles virus obtained from the culture fluid containing strains of measles virus Leningrad 16 with a titer of not less than 105.0TCD50/ml, Edmonston not less than 106.0TCD50/ml, NovO/96-not less than 108.0TCD50/ml by inactivation of its infectious activity of the detergent and the release of protein from the cell lysate by chromatographic purification of at least 2 μg/ml with a purity of not less than 70%.
Cultural vaccinated fluid obtained during separate cultivation of strains of measles virus Leningrad 16, Edmonston and NovO/96 on the monolayer culture of Vero cells with subsequent mixing of cultural vaccinated liquids in the ratio of 1:1:1 by volume).
Antigenic properties of complex antigen. Identifies all the specific antibodies to measles virus in the sera of patients and vaccinated people in higher titers than using each antigen separately (table 1).
On the basis of a complex antigen created immunosorbent is the main component of a set of reagents one-step ELISA for the detection of the antibodies is to measles virus. Reagents Anti-Measles Ig Resorbent" consists of:
- immunosorbent - complex antigen measles virus, adsorbed in the wells of polystyrene collapsible tablets;
- K1+ - positive control sample No. 1 serum human blood containing antibodies to measles virus and does not contain antibodies to the BEACH-1, HIV-2, HCV,
- Treponema pallidum and HBs-antigen, inactivated by heating at a temperature of 56°C for 3 h - 1 FL., 2 ml;
- K2+ - slabopolozhitelnym control sample No. 2 serum human blood containing antibodies to measles virus in the minimum reliably detectable quantity that does not contain antibodies to HIV-1, HIV-2, HCV, Treponema pallidum and HBs-antigen, inactivated by heating for 3 hours at a temperature of 56°C - 1 FL., 2 ml;
- - - Negative control sample serum human blood that does not contain antibodies to measles virus, HIV-1, HIV-2, HCV, Treponema pallidum and HBs-antigen, inactivated by heating at a temperature of 56°C for 3 h - 1 FL., 3 ml;
- RK - conjugate solution - a solution of monoclonal mouse antibodies to human Ig conjugated to horseradish peroxidase - 1 FL., 11 ml;
PX - Chromogen solution containing TMB 1 vial, 13 ml;
- FSB-T(×25) - 25-fold concentrate of phosphate-saline buffer solution with tween - 1 FL., 26 ml;
stop-reagent 1 vial, 6 ml.
Analytical sensitivity and analiticheskii what I specificity set were determined in ELISA with a comparative statement panel of sera, characterized in ELISA using kits for the quantitative determination of IgG antibodies to measles virus production company "Vector-best" and CJSC "Medical-biological Union" - "Octocore-IgG and Measles-IgG-DS", respectively. Comparative characteristics of the sets of reagents for one-step ELISA, including "Anti-Measles Ig Resorbent", obtained on the basis of the proposed complex antigen listed in table 2.
|Comparative characteristics of a set of reagents for one-step ELISA Anti-Measles Ig Resorbent"|
|The reagent kit||The name of the donor group sera analyzed panel|
|Not vaccinated, not ill N=89||Once vaccinated N=89||Twice vaccinated N=89||Recover from measles N=89|
|"Anti-Measles Ig Resorbent"||0/-||90/0,67||98/5,5||100/17,4|
|Note: table 2 shows the number of positive samples/average specific activity of positive specimens in IU/ml|
Table 2 shows that the analytical sensitivity and specificity of the test system "Anti-Measles Ig Resorbent", obtained on the basis of the proposed complex antigen, significantly higher than the known analogues.
Example 1. The method of obtaining complex antigen measles virus
Getting vaccinated liquid
The original Vero cells stored in liquid nitrogen and grown by serial subcultures. As the growth environment of the use of the solution Needle MEME with 8-10% of fetal serum of cattle. The cell culture is grown for 3-4 days at sowing cell concentration of 5-10×105cells/ml of medium and subcultured accepted way.
Accordingly measles virus (strains L-16, Edmonston, NovO/96) separately infect 1 litre culture vessels with monolayer culture of Vero cells (multiplicity of infection of 1:10). After incubation at 20-25°C for 40 minutes in the culture vessels add 200 ml of culture among the s Needle MEM, containing 4,0±0,1 ml bovine serum, 0.001% trypsin, 200 u/ml benzylpenicillin and 200 ng/ml of streptomycin. After incubation for 5-7 days at (36±1)°C pour nutrient medium. The titer of measles virus, strain L-16, CVG is not less than 105.0TCD50/ml, strain Edmonston not less than 106.0TCD50/ml, strain NovO/96 - not less than 108.0TCD50/ml.
Obtaining a lysate of Vero cells
Under cultivation the transfer of the virus from cell to cell is due to the formation of simpleton without departing from the main quantity in the last CVG, therefore, the accumulation of the virus occurs in CVG and inside cells. In a culture vessel with add 5-7 ml of 0.01 M Tris model HC1, pH=7.6 and the detergent Triton X-100 to a final concentration of 2-3%. The suspension is incubated for 1 h at (36±1)°C and stirring, is then treated with ultrasound 22 Hz 2 times in 20 seconds. The cell lysate is released from the cell detritus sedimentation QUE at 4-8°C for 8-12 h or low-speed centrifugation (rotor JA-14 centrifuge J2-21 ("Beckman, USA)at 10,000 rpm for 20 min at 4°C.
Example 2. Selection conditions chromatographic purification of the complex antigen
DEAE-pulp ("Whatman", Sweden) prepared according to the recommendations of the manufacturer. The column with DEAE-cellulose balance buffer 20 mm Tris-HCl, pH to 7.6. United su is pensio antigenic material three strains of measles virus to be applied on the column at a rate of 1 ml suspension of 1 ml of the sorbent. After application the column was washed with the same buffer. The elution of sorbed material is carried out by steps concentrations of NaCl in the same buffer solution to reduce the optical density at a wavelength of 280 nm in the output column with the solution to the "zero" values. Each faction examine the quality of the antigen sensitivity and specificity by ELISA (see table 3).
Example 3. Chromatographic purification of the complex antigen
The column with DEAE-cellulose balance buffer 20 mm Tris-HCl, pH to 7.6. Combined suspension antigenic material three strains of measles virus to be applied on the column at a rate of 1 ml suspension of 1 ml of the sorbent. After application the column was washed with the same buffer containing 0.2 M NaCl. The elution of the target complex antigen conduct 0.4 M NaCl in the same buffer. The target fraction is controlled to match the quality of antigen sensitivity and specificity by ELISA.
|The ELISA results in the formulation of positive and negative samples using immunosorbent made on the basis of different fractions of the chromatographic purification of complex antigen|
|The concentration of NaCl for elution fraction of the antigen with the number of the NCI||OF positive sample||OF negative sample||OP(+)/OP(-)|
|Rezorbiruetsa fraction (slippage)||0,117||0,256||0,5|
|0.1 M NaCl||0,075||0,179||0,4|
|0.2 M NaCl||0,187||0,120||1,6|
|0.3 M NaCl||1,459||0,121||12,1|
|0.4 M NaCl||2,453||0,163||16.2|
|0.5 M NaCl||1,469||0,204||7,2|
|0.7 M NaCl||0,552||0,192||2,9|
|1 M NaCl||0,340||0,184||1,8|
The output of high-purity complex antigen with 3 l KUR and lysate of cells is 15-20 mg
Example 4. Manufacturer immunosorbent assay
Immunosorbent prepared by the addition of 0.20±0.06 ml complex antigen measles virus after purification on column (see Example 3) 1,00±0,01 l R-RA sorption. The composition of R-RA sorption: 2,00±0.01 g of sodium carbonate, and 0.50±0.01 g of sodium azide, 0,400±0,012 ml of 2% phenolphthalein, purified water to 1 liter. Immunosorbent stirred on a magnetic stirrer for 1 h at a temperature of from 18°C to 24°C.
Deposition and sorption immunosorbent assay for tablets: for sorption use tablets Nunc Medisorp - "Nunc a/S, Denmark, or similar. Filling volume holes - 100-105 mm. Sorption was carried out at a temperature from 17°C to 27°C, time of sorption - 18-20 hours
Example 5. The method of use of a set of reagents for one-step ELISA Anti-measles Ig Resorbent"
For the analysis used the serum or plasma of human blood. For analysis only 10 μl of the sample. For the detection of antibodies of class G to the measles virus, you can use the serum (plasma) blood of the person as freshly prepared and not stored for more than 7 days at a temperature of from 2°C to 8°C in the absence of microbial contamination or not more than 12 months at a temperature of minus 20°C. freeze serum or plasma to a temperature of minus 20°C not more than two times.
For the reaction to all wells contribute 90 ál R-RA is svedeniya sample (composition: 2,00±0.01 g of sodium carbonate, 0,50±0.01 g of sodium azide, 0,400±0,012 ml of 2% phenolphthalein, purified water up to 1 liter). The hole in the tablet A1 contribute 10 ál K1+in wells B1 and C1 make 10 ál K2+. In well D1 contribute 10 ál To. In the remaining wells contribute 10 µl sample serum (final dilution of specimens and controls - 10 times). The solution is mixed by pipetting and incubated for 40 min at 37°C. after incubation the contents of the wells is removed and the plate washed seven times working solution FSB So In every hole making 100 ál of R-RA chromagen and the plate is placed for 15 min in the dark place at a temperature of 37°C. the Reaction is stopped by introducing into each well of the tablet in 50 µl of stop solution.
The value of OP solutions in the wells immunosorbent assay measured on the spectrophotometer in a two mode: main filter 450 nm, reference filter in the range from 620 to 700 nm. The results of the test is considered positive if the OD test serum more APK+Ms. the lower limit of positive values corresponds to 0.28 IU/ml). The results of the test is considered negative if the OD test serum less OPK+cf - 20% (upper limit negative values corresponds to 0.12 IU/ml).
Psiv. calculated by the formula:
where Opseu. - the OD value of the test serum.
Asyv. (the titer of antibodies in IU) R is schityvat by the formula:
where a and b are constants specific to each series specified in the Instructions that accompany each set).
An example of the calculation data obtained are given in table 4.
|An example of the calculation of the titer of antibodies to measles virus in the test serum|
|(APK+AVG) is the average OD value in the wells A1 and B1||2,919 - range Applications|
|(APK+AVG) is the average OD value in the wells C1 and D1||(0,273+0,275):2=0,274 - range of Application|
|The lower boundary of the positive values||>(APK+cf)=0,274|
|The upper boundary of the negative values||<(APK+cf)-20%=0,220|
|The OCM - value OP in well E1||0,056<(APK+cf)-20%=0,220|
|The above values indicate the results of ELISA|
|Opsys. - the OD value of the test serum||1,920|
|and - the value of the constant Application||1,4|
|b - the value of the constant Application||2,1|
|(Psiv. - a):b||(1,752-1,4):2,1=0,168|
|Response: the specific activity of the serum 1,47 IU/ml|
Example 6. Using a set of reagents "Anti-Measles Ig Resorbent" on the basis of the inventive antigen in clinical practice
1. During outbreaks of infectious diseases, Blagoveshchensk in 2010, a set of reagents "Anti-Measles Ig Resorbent" was used for serological confirmation of clinical diagnosis of measles". In 2 patients 2-3 days after the onset of clinical signs of disease and in 2-4 weeks spent drawing blood and obtaining sera for serological confirmation of the diagnosis of measles." Increasing titers of specific antibodies in paired sera studied in ELISA using commercial kits "Melisa Measles-IgM production CJSC "MBS" and in parallel with experimental series "Anti-Measles Ig Resorbent", the main component of which is immunosorbent on the basis of a complex antigen measles virus. The diagnosis of measles" confirmed the Ali after PCR with specific primers for C-terminal part of the gene encoding the NP protein of measles virus, and sequencing amplification of the fragment.
The set of "Melissa Measles-IgM revealed specific IgM antibodies to measles virus in both patients. When setting ELISA kit Anti-Measles Ig Resorbent" these patients also noted the presence of specific IgM antibodies to measles virus. When determining the rise in the titer of specific antibodies in paired sera with Anti-Measles Ig Resorbent" in both patients was zaregistrirovan at least 4-fold increase of specific IgG antibodies to measles virus.
In samples of patients by PCR was detected genetic material (RNA) of the measles virus. Sequencing revealed the identity of the virus that caused the outbreak, to genotype N.
Thus, the set of reagents "Anti-Measles Ig Resorbent" can be used for early detection of antibodies to measles virus, as well as for the detection of specific antibodies in paired sera obtained from patients, and thus to identify, remove or confirm the diagnosis in the presence of a disease with clinical manifestations characteristic of measles.
2. In the period from 2005 to 2010, in the Novosibirsk region was carried out to study the titre of specific antibodies to measles virus in the blood serum of children age 6 years prior to the second vaccination against measles. The titers of specific antibodies was studied in the reaction of rtga (by the conventional method using erythrocytes monkeys Macaca mulatta) and ELISA (using the proposed set of reagents "Anti-Measles Ig Resorbent").
The results of the comparison of the ELISA and rtga to determine measles antibodies in the serum of children showed that using "Anti-Measles Ig Resorbent" specific antibodies were identified 235 sera from 237 investigated sera. A similar result (235 sera from 237) were obtained using rcga. Simple averages of the specific activity of measles antibodies defined by a set of "Anti-Measles Ig Trearment were higher than specified in RTCA (6,2 IU/ml and 3.3 IU/ml, respectively).
Thus, the set of reagents for one-step ELISA Anti-Measles Ig Resorbent" can be used to determine the strength of individual immunity to the disease "measles".
1. Complex antigen measles virus used as a component of an enzyme immunoassay system for the diagnosis of antibodies to measles virus obtained from the culture fluid containing strains of measles virus Leningrad 16 with a titer of not less than 105.0TCD50/ml, Edmonston not less than 106.0TCD50/ml, NovO/96 - not less than 108.0TCD50/ml by inactivation of its infectious activity of the detergent and the release of protein from the cell lysate by chromatographic purification of at least 2 μg/ml with a purity of not less than 70%.
2. Complex antigen according to claim 1, characterized in that the culture of the other vaccinated fluid obtained during separate cultivation specified in claim 1 strains of measles virus in monolayer culture of Vero cells with subsequent mixing of cultural vaccinated liquids in the ratio 1:1:1 by volume.
FIELD: chemistry; biochemistry.
SUBSTANCE: invention concerns medical virology and microbiology. Strain is deposited in culture collection of Federal State Research Institution State Research Centre of Virology and Microbiology "Vektor" of Rospotrebnadzor, under registration No VB-05. Strain features higher productivity. More sensitive immunoenzyme test system for hepatitis virus antibody diagnostics is created on the basis of this strain. Invention can be applied in virology.
EFFECT: production of more sensitive immunoenzyme test system for hepatitis virus antibody diagnostics.
2 cl, 1 dwg, 1 tbl, 4 ex
FIELD: medicine; ophthalmology.
SUBSTANCE: invention is intended for simultaneous verification of uveal melanoma and forecast of metastasis development. Immunohistochemical analysis of protein S-100 and melanin-A expression within tumour cell is accompanied with simultaneous estimation of S-100-positive cells number within visual field. Uveal melanoma is verified if reaction of S-100 and melanin-A is positive with forecasted high possibility of tumour dissemination in case number of S-100-positive cells number is less than 50.
EFFECT: simplified examination associated with high specificity and sensibility of simultaneously verified uveal melanoma and forecasted metastasis.
1 tbl, 1 dwg, 2 ex
FIELD: medicine, veterinary.
SUBSTANCE: specific antibodies to virus of goose enteritis are determined by IEA where virus received following injection of epizootic strain "P-75" of enteritis virus, is used as an antigen, and macro porous glass with diameter not less than 700 is used for purification of virus-containing material, the goose Ig G specific immunoperoxidase conjugate is used as an anti-specific agent, and the IEA results are calculated by use of the formula: titer = antiIg[2.02(lg S/P)+3.51], where S is for optical density of tested serum; P is for optical density of the positive control.
EFFECT: method reduces the test time and economical costs.
2 tbl, 2 ex
SUBSTANCE: method involves determining total IgA quantity using immunoenzyme method based on polyclonal IgA antibodies as binding antibodies on the micropanel and the same antibodies in conjugate with peroxidase for detecting bound IgA in sample before and after treatment with specific IgA1-protease. Total IgA1 and IgA2 content is determined before being treated with enzyme. IgA2 content only is determined after being treated with enzyme. IgA1 is not determined after being disintegrated with IgA1-protease. Kit has microplate containing absorbed polyclonal IgA antibodies, conjugate of the same antibodies with peroxidase, reference sample having given IgA concentration, IgA1-protease and substrate buffer.
EFFECT: simplified method and kit for determining IgA1 and IgA2; improved results reproducibility; no specific monoclonal antibodies being used.
2 cl, 1 tbl
SUBSTANCE: method involves absorbing myeloma IgA1 on micro panel, incubating solutions containing IgA1-protease and determining enzyme activity from IgA1 sorbate quantity reduction determined by conjugate with polyclonal IgA antibodies peroxidase. Determining IgA1-protease activity with inhibitors available in various concentrations enables one to calculate its inhibition constants. Kit has microplate containing absorbed myeloma IgA1 preparation, peroxidase conjugate with antibodies against human IgA and substrate buffer.
EFFECT: simplified method and kit for determining IgA1-protease activity and studying its inhibition process.
2 cl, 1 dwg, 1tbl
FIELD: veterinary virology.
SUBSTANCE: the present innovation deals with interaction of antibodies with an antigen, with antibodies labeled with horseradish peroxidase, addition of substrate mixture and registration of reaction results. Moreover, one should apply plotting boards with presorbed antibodies and general immunoenzymatic conjugate. The innovation enables to shorten terms for diagnosis and obtain more significant results of diagnostics.
EFFECT: higher efficiency.
FIELD: biotechnology, analytical chemistry.
SUBSTANCE: claimed method includes providing of complexes between antigen molecules and specific antibodies on carrier surface, wherein said complexes are disclosed by addition of enzyme label thereto followed detection thereof based on formation of enzyme reaction product. Enzyme label addition is carried out by two protein interaction, namely bacterial ribonucleaze barnase and barstare, which is inhibitor thereof, wherein either of the two is added to immunoreagent, and the other one is added to enzyme label. Abovementioned complexes have high affinity, specificity, and binding constant of 1014 M-1.
EFFECT: new method for antigen detection.
6 dwg, 2 ex
FIELD: veterinary and medicine.
SUBSTANCE: invention, in particular, relates to production and use of biological preparations intended for differential diagnostics of brucellosis and to a method of differentially diagnosing brucellosis. Method involves serologic analysis of sera using antigenic "IFK", which is horse-radish peroxidase-labeled electrophoretically purified polypeptide fraction of virulent strain B.arborus 54. Diagnosis of brucellosis is stated when anti-brucellosis antibodies in sick animal sera diluted to 1/100 and higher are revealed at CSP reaction intensity 2.1 and higher.
EFFECT: elaborated method increasing immuno-enzymatic test specificity and allowing performing differentiation of postvaccinal immunological reactions and postinfectious reactions induced by microorganisms having antigenic affinity with brucellas, especially Yersinia enteriocolitica.
2 cl, 4 ex
FIELD: medicine, biotechnology.
SUBSTANCE: the present innovation deals with elaborating diagnostic reagents for testing prionic protein in mammalian cerebral tissue due to IEA technique and refers to antibodies specifically reacting with prionic protein PrP or its fragment. The innovation includes polyclonal antibodies obtained to synthetic peptides including amino acid sequences of bovine prionic protein 143-168, 101-134 and 211-241, their conjugates with horseradish peroxidase and diagnostic reagents obtained upon their basis that enable to detect prionic protein in mammalian cerebral tissues.
EFFECT: higher specificity of detection.
15 cl, 8 ex, 8 tbl
FIELD: medicine, obstetrics.
SUBSTANCE: in pregnant women at pregnancy terms being after 39 wk in average portion of morning urine one should detect due to a solid-phase immunoenzymatic assay the concentration of pregnancy-associated protein-A (PAPP-A). At the level of PAPP-A being 768.2 ng/ml and more it is possible to conclude upon physiological mature pregnancy. The innovation is noninvasive and enables to increase accuracy in predicting true over-mature pregnancy.
EFFECT: higher efficiency and accuracy of diagnostics.
2 ex, 1 tbl
SUBSTANCE: group of inventions is referred to the area of medicine, namely to the area of virusology, and is related to virosomes containing hemagglutinin extracted from influenza virus produced in the cell lines, compositions, containing said virosomes, means of manufacturing and applications. The essence of the invention including virosomes containing hemagglutinin extracted from influenza virus produced in the bird cell lines, compositions, containing said virosomes, application of virosome as a vessel, set, methods of vaccination, methods of treatment and methods of virosome production.
EFFECT: production of virosomes having improved merging capability and increased immunogenicity.
21 cl, 3 tbl, 5 dwg
SUBSTANCE: what is described is a composition containing an ordered antigen pattern where antigen represents IL-1, mutein IL-1 or fragment IL-1. There is also offered a based vaccine. The compositions offered in the invention can be applied for producing vaccines for inflammatory diseases and chronic autoimmune diseases, transmittable diseases and cardiovascular diseases.
EFFECT: compositions effectively induce immune responses, particularly humoral immune responses; compositions are the most suitable for effective induction of autogenic immune responses.
46 cl, 2 dwg, 3 tbl, 14 ex
SUBSTANCE: strain was isolated from the pig which died in LLP "Gorkaya Balka" in Soviet district of Stavropol region during disease epizootic in 2008. Strain is new, earlier unknown, isolated on the territory of RF and is designated as strain "Stavropol 01/08". It possesses high infectious activity, accumulates in cultures of bone marrow cells of pigs and pigs' leukocytes in titre 6.0-7.0 lg HAU50 cm3. Virus titre on pigs constitutes 7.0-7.5 lg LD50 cm3. Death of infected pigs occurs with symptoms characteristic of acute form of ASP 3-6 days after manifestation of clinical symptoms of disease. Invention can be used in research institutions and diagnostic centres as reference-strain in carrying out virological, molecular-genetic and monitoring research.
EFFECT: high infectious activity of strain.
3 tbl, 3 ex
SUBSTANCE: species-specific virulent strain of bacteriophage Acinetobacter baumannii AP22 of Myoviridae family is isolated from clinical material and deposited in collection of microorganisms museum FSIS "State Research centre of applied microbiology and biotechnology" under number Ph-42. Bacteriophage possesses expressed lytic activity, lyses 68% of A. Baumannii strains, isolated from clinical material, and is used for identification of microorganisms opf said species in bacteriologic analysis of clinical material, as well as for elaboration of complex medications against A. baumannii-infections.
EFFECT: invention ensures wide spectrum of activity within said species.
2 dwg, 1 tbl, 4 ex
SUBSTANCE: strain "АС-21/07" of Aujeszky's disease virus was deposited in collection of VGNKI under registration number "АС-21/07"-DEP. Strain is obtained from original strain MK-25 by selection. In culture of cells PSGK-60 and PSGK-60S virus 19-20 passage is accumulated to 8.0-8.5 lg TCD50/cm3.
EFFECT: strain is attenuated for pigs, sheep, rabbits and possesses high biological, antigenic and immunogenic activity in native form and after inactivation.
2 dwg, 3 tbl, 4 ex
SUBSTANCE: strain of bird flu of A/Herring gull/Mongolia/454/08/ H13N8-subtype was deposited in Collection of microorganisms of Federal State Institution of Science "State research centre of virology and biotechnology "Vector" Rospotrebnadzor under registration number V-436 and is used in obtaining antigen-containing substrate and serum for diagnostic purposes. Claimed strain can be used for serodiagnostics of influenza of H13-subtype in RHAI, as component of panel of polyclonal serums to different subtypes of influenza A virus (H1-16) for typing versions of influenza virus newly isolated from nature in reaction of hemagglutination inhibition (RHAI), as well as for identification of antibodies to virus of H13-subtype influenza in serums of wild animal blood for estimation of population immunity to virus of said subtype influenza in RHAI.
EFFECT: improvement of strain properties.
2 tbl, 3 ex
SUBSTANCE: group of inventions refers to veterinary science, and concerns inactivated chimeric vaccines and related methods of application. Substance of the inventions involves an inactivated chimeric flavivirus containing a first flavivirus representing a yellow fever virus in which nucleotide sequences coding pre-membrane and membrane proteins are substituted by nucleotide sequences coding pre-membrane and membrane proteins a second flavivirus representing a West Nile fever virus. The inventions also involve an immunogenic composition and a vaccine for protection of animals from the flavivirus infections, involving the inactivated chimeric flavivirus.
EFFECT: development of the advanced immunogenic composition and the vaccine on the basis of the inactivated chimeric flavivirus.
14 cl, 3 ex, 5 tbl
SUBSTANCE: nucleic acid contains a gene segment of a influenza virus and a bacteriophage polymerase promoter or a complementary chain of said nucleic acid. What is described is a composition containing a cell or a material produced of a cell according to this invention, or a virus, or a material produced of a viral particle according to this invention. The invention can be used in medicine.
EFFECT: nucleic acid allows producing replicative viral particles without the use of an assistant virus.
26 cl, 6 dwg, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to field of biotechnology of veterinary medications. Vaccine against rabies virus represents allantoic fluid of chicken embryos. Fluid contains glycoprotein of rabies virus, as well as mixture if recombinant adenoviruses of birds, which carry gene of surface glycoprotein of rabies virus, one of which contains secreted form of surface glycoprotein of rabies virus, and second - membrane-bound one. Interaction of glycoproteins with animal organism is reached by peroral introduction of medication.
EFFECT: invention can be applied in veterinary.
3 dwg, 1 tbl, 6 ex
SUBSTANCE: strain is produced by cross breeding of sterile equine influenza virus A/horse/Prague/1/1956(H7N1) and cold-adapted vaccine strain A/17/California/09/38(N1) on the basis of an attenuation donor A/Leningrad/134/17/57(H2N2) and contains neuraminidase of pandemic influenza virus A//California/07/09(H1N1) and hemagglutinin of equine influenza virus A/horse/Prague/1/1956(H7N7). The strain RN1/09-swine A(H7N1) is deposited in the State collection of viruses of Institution of Russian Academy of Medical sciences of D.I. Ivanovsky Institute of virology of Russian Academy of Medical Science, No. GKV 2473 and can be applied for influenza virus neuraminidase N1 antibody assay.
EFFECT: higher assay accuracy.
2 dwg, 4 tbl, 1 ex
SUBSTANCE: claimed group of inventions relates to medicine, namely to oncology, and can be applied for treatment of neoplasm, as well as for producing medication for it treatment. For this purpose virus of Newcastle disease in combination with gluoropyrimidine and camptothecin compound during one or more cycles in total quantity, effective for subject treatment.
EFFECT: application of claimed inventions results in stable regression of tumour due to synergetic anti-tumour effect from application of all three agents.
30 cl, 4 dwg, 7 tbl, 5 ex