Method of detecting provirus of cattle leukosis
SUBSTANCE: claimed invention relates to the field of biotechnology and deals with a method of detecting a provirus of cattle leukosis. The characterised method includes the detection of LTR (Long Terminal Repeat) fragment - a sequence of the leukosis provirus. Determination of the size of an amplified fragment of a nucleotide sequence by the electrophoretic separation in an agarous gel is performed. As primers applied are oligonucleotides: BL1.F 5-GAGTTAGCGGCACCAGAAGC-3 and BL1.R 5-ATAGAGCTCGCGGTGGTCTC-3. The product of synthesis constitutes 175 pairs of the nucleotides. A genomic DNA is extracted by a sorbent method with the following elution in TE buffer, amplification is carried out in the mode: 95°C for 3 minutes 1 time, 94°C for 20 seconds - denaturation, 66°C for 20 seconds - annealing of the primers, 74°C for 25 seconds - elongation, 45 times, 74°C for 3 minutes - chains completion. As a positive control of reaction proceeding applied is a preparation of a positive control of BLV antigen.
EFFECT: invention can be applied in the molecular-genetic diagnostics of animal diseases and scientific research in veterinary.
2 dwg, 1 tbl
METHOD of DETECTING DNA PROVIRUS LEUKEMIA
The present invention relates to the field of biotechnology, molecular genetic diagnosis of animal diseases, research in veterinary science, molecular biology.
The bovine leukemia is a chronic infectious disease that is common worldwide. Called disease virus BLV (Bovine Leukemia Virus) and belongs to the genus Deltaretrovirus. By nature, it is similar to the virus HTLV and affects the bulk of B-lymphocytes. The virus leukemia cattle cause significant economic damage to the livestock farms of the Russian Federation.
In the complex of measures on improvement of herds of cattle from leukemia, the leading position belongs to the diagnosis. Issue timely and effective diagnosis of BLV in connection with extreme unfavorable epizootic situation on this disease and the lack of means of specific prophylaxis remains a critical issue.
One of the first and the main forms of diagnosis are the traditional methods of serological diagnosis of BLV (RIA, ELISA), which is not always possible to identify infected animals, and in some cases are unacceptable, for example in the early diagnosis of calves from seropositive parents that require the use of them in combination with methods mol�about genetic, in particular PCR, with which is possible early detection of BLV in dysfunctional leukemia farms, as well as increasing the efficiency and reducing the period of the anti-leukemic activities.
Polymerase chain reaction (PCR) is an effective and highly sensitive method, which is based on a multiple increase (amplification) of the studied fragment of the DNA sequence (target DNA) to the ability of its identification in gels or by using fluorescent dyes in real-time.
The specificity of the methods (NDP) is in the selection of oligonucleotide primers-primers, forward F and reverse R primers that define the scope elongation thermophilic polymerase. It is also important the selection amplificative fragment of proviral DNA of bovine leukemia.
The known method (Patent - US 6646116 B1, 11.11.2003, C07/H 21/04) differentiation gene variant TAX virus Laosa cattle, consisting in carrying out a panorama using primers capable of identifying the full sequence of the gene for further detection of mutations encoded by this gene amino acid sequences and identifying options for the TAX gene of BLV. In particular we used the following primers:
In tax2:5 AG TST AGA GCT GTC GAC TST GTC TG 3
In tax3:5 ACC TCG AGA TGG CAA GTG TTG TTG GTT GG 3.
Also known a method of detecting DNA provirus leukemia� cattle method pnrm. in the mode of real time (RT-PCR) and nested pnrm (nested PCR) (Log - Journal of Veterinary Diagnostic Investigation (J Vet Diagn Invest). 2003, Vol.15, 72-76. USA).
The proposed primers complementary to the POL gene have the following structure:
To use the panorama in real time together with primers to add to the reaction mixture was additionally synthesized POL probe (molecular beacon) 5 - (FAM)-
fluorescent label, which is the detection signal.
The method of nested PCR (nested PCR) involves the use of two pairs of primers - outer (1 round) and nested (2nd round). When both methods PCR product size is 202 nucleotide pairs. However, the known methods of detecting DNA of BLV provirus: in the analogue did not provide a detection area conservative Pol gene provirus.
A method of detecting DNA of bovine leukemia provirus Pol gene using the primers: RA 2: 5 - TGA ACG GAC AAA TGG ACT GCT C-3, P r2 5 - CCG ACA GAG AGC GAG GAG AG -3, giving the output product size 438 nucleotide pairs with subsequent visualization in 1.5% agarose gel in the presence of ethidium bromide as a dye (patent-2445370).
The drawback of all these methods is the requirement of a minimum number of copy number of sequences in the analyzed volume equal to 10 copies. The disadvantage is also the fact that while on study� persistent lymphocytosis titer of virus in the blood falls and may reduce the number of provirus copies below the claimed analytical sensitivity, which leads to false-negative results.
The object of the invention is to develop an effective method of detecting DNA provirus of bovine leukemia by PCR, allows for the increase of sensitivity for a selected region of DNA to detect the presence of the provirus in the diagnosis of Ural type BLV, with the ability to detect DNA provirus of bovine leukemia in the presence of a minimum number of copies of sequences in the analyzed volume (less than 10 copies), as well as enabling analysis at the stage of persistent lymphocytosis, at a lower titer of virus in the blood.
The task is solved in that using a straight F and reverse R primers for the detection of a fragment of the LTR (Long Termal Repeat) sequences of provirus bovine leucosis, amplification is carried out (building up) of the desired fragment, and determining the size amplificatoare fragment of the nucleotide sequence is carried out electrophoretic separation in an agarose gel, the primers are the oligonucleotides of the following structure: BL1.F 5-GAG TTA GCG GCA CCA GAA GC-3; BL1.R 5-ATA GAG CTC GCG GTG GTC TC 3, and the product of the synthesis of primers is 175 pairs of nucleotide and genomic DNA isolated sorbenty method, followed by elution in buffer TE (simple rapid method able� " s DNA), and amplification is carried out in solution in 80 mm Tris-HCL (pH 8.0), 0.1% Triton X-100, 24 mm (NH4)2SO4, 0.5 mm EDTA, 0.2 mm of each of the triphosphates, 0.5 mm of each primer and 1 unit of TAG polymerase and 40 ng of the analyzed genomic DNA of an infected animal, when the final volume of 20 µl solution, wherein the amplification is carried out in the regime: 95°C 3 minutes 1 time (process denaturation), 94°C - 20 seconds denaturation, 66°C - 20 seconds - annealing of primers, 74°C - 25 seconds - elongation, 45 times, completing circuits 74°C - 3 minutes, moreover, the detection of amplified fragments is carried out in 2% agarose gel in buffer DIDN, 0.5 mm with ethidium bromide, and as a positive control of PCR using the drug positive control antigen BLV.
Positive analysis of the confirm test systems domestic manufacturers.
In the process of the carried out works as a DNA target selected regions of the LTR (Long Terminal Repeat). The LTR region is found in all retroviruses and is required for integration of the provirus into the host genome region fenceroy provirus genome, thus increasing chopinot DNA target 5 two times, respectively, increases the sensitivity of the method that shows the obvious effect of the proposed method.
Then using the computer program Clustal W2 multi sequence alignment based on inform�tion is represented in the database of Internet resources NCBI GenBank were selected and analyzed 50, in our opinion, the most discordant genotype BLV to determine conservative areas of the virus. The selection of primers was performed using the program Lasergene V7.1 (USA). When conducting a computer analysis to the selected parcel LTR define basic criteria: a) the degree of homology of primers; b) lack semicomplete sites; b) the proximity of the melting temperature of the primers. We were picked up the following primers:
for the detection of LTR sequences of bovine leukemia provirus with the release of the product length 175 nucleotide pairs. Synthesis of oligonucleotide primers were ordered in the commercial center.
Fig. 1. presents the èlektroforegramme of amplification products M - token increments of 50 nucleotide pairs (50-500) " - "- healthy animals, " BL "+" - positive leukemia virus animals, With "-" and " + " for negative and positive controls.
Fig. 2. presents the èlektroforegramme of amplification products; M - marker increments of 50 BP (50-500), " BL "+" infected animals detected by the developed method, BLV "+" infected animals diagnosed using a diagnostic system of the Russian manufacturer, BIV test the specificity of the primers in the presence of virus DNA immunodeficita cattle, With "-" and "+" negative and positive controls.
For �technical confirmation of the possibility of early detection of bovine leukemia provirus in blood of infected animals were selected for the group.
A. Animals with an acute form of leukemia identified hematological method 28 goals.
Selected as the positive group.
B. a Group of youngsters at the age of 2-3 months after birth and is diagnosed on the subject of BLV infection by the method of REID (reaction immunodiffusion) showing a positive result.
Animals were selected on farms farming "Isimgready" Ishim district of the Tyumen region.
Both groups of subjects were studied by the method of the DAG, the data presented in table 1.
Animals in group B the number 4, which showed a negative result by PCR diagnosis were isolated and separated from the rest of the herd. Subsequent test analysis after 6 months confirmed prior and was negative. The presence of REED + result in young was the result of the influence kolostralnogo immunity twice and confirmed the test result pnrm analysis is considered in this case as the most accurate and informative method for early diagnosis of bovine leukemia provirus in calves.
This method is accurate, sensitive, with which you can more accurately in the early stages to identify a fragment of proviral DNA of leukemia virus of cattle in the biological material, in the presence of minimalno� the number of copies of the sequence in the analyzed volume at the stage of persistent limpieza, at low titer of virus in the blood.
A method for the detection of provirus bovine leucosis, including the identification of the fragment of the LTR (Long Terminal Repeat) sequences of leukemia provirus, determining the size of amplificatoare fragment of the nucleotide sequence by electrophoretic separation in agarose gel, the primers are the oligonucleotides of the following structure: BL1.F 5-GAGTTAGCGGCACCAGAAGC-3; BL1.R 5-ATAGAGCTCGCGGTGGTCTC-3, and the product of synthesis is 175 pairs of nucleotides, wherein the genomic DNA is isolated sorbenty method, followed by elution in buffer TE, and amplification is carried out in a solution of 80 mm Tris-HCL (pH 8.0), 0.1% Triton X-100, 24 mm (NH4)2SO4, 0.5 mm EDTA, 0.2 mm of each of the triphosphates, 0.5 mm of each primer, 1 unit of TAG polymerase and 40 ng of the analyzed genomic DNA of an infected animal, at finite solution volume of 20 μl amplification is carried out in the regime: 95°C 3 minutes 1 times, 94°C - 20 seconds denaturation, 66°C - 20 seconds - annealing of primers, 74°25 seconds elongation, 45, 74°C - 3 minutes - building up of the circuits, and detection of amplified fragments is carried out in 2% agarose gel DIDN buffer, 0.5 mm with ethidium bromide, and as a positive control of PCR using the drug positive control antigen BLV.
SUBSTANCE: invention relates to biochemistry. Described is method of detecting presence or absence of several serotypes of human papillomavirus (HPV) in biological sample by means of multi-channel analysis system, where said analysis system detects larger quantity of serotypes than the number of existing channels of detection. Method peculiarity consists in the fact that first and second sets of primers and probes, which are degenerate with respect to each other, are used; as well as the fact that each degenerate probe has signal grouping, each of which studies signal, detected on one and the same channel. Third set of primers and probes is not degenerate with respect to two other sets of primers and probes and is distinctive for third serotype, which does not belong to serotypes, to which mainly annealing of degenerate sets of primers and probes takes place, and where third set of primers and probes has signal grouping, which emits signal with wavelength, which is the same or differs from wavelength, emitted by signal grouping of degenerate probes. Respective primers and probes are presented.
EFFECT: invention makes it possible to detect larger quantity of HPV types than the number of existing channels of detection in analysis system.
11 cl, 3 dwg, 6 tbl
SUBSTANCE: inventions relate to the field of DNA-genealogy and deal with a method of determining haplogroups of human Y-chromosomes, a test-system and oligonucleotide primers. The characterised method is realised in two stages. At the first stage genetic typing is carried out by basic haplogroups R,N,J,I,Q,C,E,D,G,O of an Y-chromosome tree by multiplex PCR with the application of specific oligonucleotide primers of the first and second sets of the test-system. At the second stage, if mutation is detected, additional multiplex PCR for typing by subhaplogroups is carried out. If at the first stage mutation is not detected, multiplex PCR for typing by rare haplogroups A,C3,F,H,K,L,O3 and T is carried out. The area of annealing primers outside mutation zones within the limits of species specificity corresponds to sequences from the first set of the test-system. The area of annealing of the oligonucleotide primers, directly adjoining the mutation zone within the limits of species specificity corresponds to sequences from the second set of the test-system.
EFFECT: inventions can be applied for the determination of haplogroups of the human Y-chromosome.
3 cl, 2 dwg, 7 tbl, 2 ex
SUBSTANCE: DNA is recovered from peripheral venous blood which is followed by a genetic typing of the APOE gene and detecting polymorphous alleles APOE*2, APOE*3, APOE*4; if the assays shows any genetic types containing alleles APOE*2, a high risk of endometrial cancer is predicted.
EFFECT: invention provides a highly specific criterion for predicting the risk of hyperproliferative diseases of the endometrium, including endometrioid adenocarcinoma in females with hyperplastic processes in the endometrium.
2 dwg, 10 tbl, 4 ex
SUBSTANCE: invention refers to medicine, namely to a method for the prediction of developing occupational hyperkeratosis. Substance of the method consists in recovering DNA from peripheral venous blood lymphocytes, genotyping TP53 gene rs1625895 polymorphism by polymerase chain reaction with restriction enzyme digest analysis. Finding G/A genotype and A allele ensured predicting a risk of developing occupational hyperkeratosis.
EFFECT: using the invention enables predicting developing occupational hyperkeratosis after the body has been exposed to occupational hazards.
2 tbl, 2 ex
SUBSTANCE: presented group of inventions refers to medicine, molecular biology and biotechnology. A method for determining lung cancer cell sensitivity to cisplatin involving determining MLH1, ERCC1, DDB2, AKR1B1, FTL genes expression patterns vs. cisplatin IC50 for known cell lines; constructing cisplatin IC50 for these cell lines vs. derived gene expression patterns calibration straight, and hybridising on a microchip. What is also presented is a set of oligonucleotide probes presented by SEQ ID NO: 9-13.
EFFECT: presented group of inventions provides effective aids and methods for determining lung cancer cell sensitivity to cisplatin.
2 cl, 1 dwg, 2 tbl, 3 ex
SUBSTANCE: invention relates to field of biochemistry, in particular to method of identifying changes in gene expression, characteristic of ageing, in selected tissue. Selected tissue represents cardiac, muscular, cerebral or adipose tissue. Method includes selection of one or several genes, differentially expressed in tissue of old subjects in comparison with young subjects, with application of two criteria. Said criteria are: change of expression in at least 50% of lines, breeds or ethnic groups of tested species at preliminarily determined significance level p<0.10, as well as at least partial reversion of changes in gene expression by restriction in caloric content.
EFFECT: invention provides obtaining reliable tissue-specific biomarkers of ageing.
7 cl, 11 tbl, 5 ex
SUBSTANCE: invention refers to molecular biology and can be used in diagnosing cardiomyopathies of various origin Presented is a set of synthetic oligonucleotides for identifying mutations of a coding part of desmin (DES) gene associated with cardiomyopathies. The mutations are identified by detecting a full sequence of the coding part of DES gene. The coding part of DES gene is amplified by means of 7 pairs of synthetic oligonucleotides at the same temperature and annealing time, and the prepared amplification products are sequenced by means of one pair of universal primers.
EFFECT: presented invention enables the sensitive and specific detection of DES gene mutations, with reducing the amplification reaction time, the number of manipulations, the time of agent application for the sequencing reaction, and reducing a probability of a reaction error.
3 cl, 1 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology, more specifically to AXL signalling pathway inhibitors, and can be used in medicine. What is produced is a soluble AXL polypeptide version free from AXL transmembrane domain, which contains at least one amino acid modification in position No. n, wherein n is specified in 32, 72, 87, 92 or 127 or a combination thereof, wherein n+7 is described by numbering SEQ ID NO: 1 that are wild-type AXL sequences, wherein the above modification increases a AXL polypeptide binding affinity to protein 6 specifically inhibiting the growth (GAS6), which is twice as strong as the wild-type AXL polypeptide affinity. The polypeptide can be fused with Fc fragment and used in a method of treating, reducing or preventing tumour dissemination and invasion in a mammalian patient.
EFFECT: invention enables inhibiting the AXL/GAS6 signalling pathways effectively.
8 cl, 15 dwg, 6 tbl, 3 ex
SUBSTANCE: invention relates to a set of oligonucleotide primers and fluorescently-labelled probe for identification of Burkholderia pseudomallei and differentiation of the actinobacillus mallei by the method of polymerase chain reaction with fluorescence detection.
EFFECT: invention enables in a short time with high sensitivity and specificity to detect the melioidosis agent and differentiate it from the actinobacillus mallei in samples of pure cultures and biological material.
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SUBSTANCE: invention refers to medicine, molecular biology and biotechnology. What is presented is a method for determining the polymorphism of human GP6 gene coding glycoprotein VI by the polymorphous position rs 1613662 based on recording melting curves with fluorescence-labelled allele-specific oligonucleotide tests.
EFFECT: due to the more reliable determination of variable positions and a possibility to detect in one test tube with the use of standard equipment, the method can be effectively used to diagnose the individual's inherited predisposition with the recording the real-time PCR results.
SUBSTANCE: group of inventions relates to cell lysis. A method for selective lysis of animal cells and a device for detecting microorganisms are disclosed. A method for selective lysis of animal cells in a water sample with animal cells, containing or possibly containing microorganisms, includes steps of providing a water sample with animal cells, containing or possibly containing microorganisms, adding to said sample a nonionic detergent and a buffer solution to obtain a solution with pH of about 9.5 or higher, incubating said solution for a period sufficient for lysis of animal cells. The device for detecting microorganisms in the sample consists of a lysis chamber for receiving a water sample with animal cells, a vessel with an alkaline buffer solution with pH 9.5 or higher and a nonionic detergent, or a vessel with an alkaline buffer solution with pH of about 9.5 or higher and a vessel with a nonionic detergent, a filter connected to the lysis chamber for filtering the sample after lysis of the animal cells, an indication chamber for analysing presence of DNA of microorganisms.
EFFECT: present inventions enable to process a sample without considerable dilution thereof and without using enzymatic breakdown of DNA or heat treatment, and also enables to process considerably larger sample volumes and determine lower concentrations of pathogenic microorganisms in a sample.
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SUBSTANCE: characterised method comprises carrying out of PCR with use of specific primers to genes vc0497, vc0502 and vc0514 from the island of pandemicity VSP-II. The characterised test system comprises the components for isolation of DNA, the components for carrying out PCR, including, in particular, a primer mix VSPIIreg-F - 5'-TGGAAAGAAGAGCGTTACTGC-3', VSPIIreg-R - 5'-CCCTGTTGATGATGTGATTTG-3' to the gene vc0497, VSPIIpilin-F - 5'-CTGTGATTCGGGCTTTATCGG-3', VSPIIpilin-R - 5'-GCGTAAACTGAGCCAATAAGC-3' to the gene vc0502, VSPIIchem-F - 5'-CTTGATGGAGCGGAGAAAAC-3', VSPIIchem-R - 5'-CGATGAATAGCCTGTTGAAC-3' to the gene vc0514, taken in the ratio 1:1:1:1:1:1, respectively.
EFFECT: inventions enable to differentiate quickly and reliably the toxigenic genetically modified strains to genovariants with low and high epidemic potential.
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SUBSTANCE: urogenital mycoplasmosis is diagnosed by detecting Mycoplasma hominis in the clinical material, as well as for semi-quantitative determining the titre of the pathogen. The nutrient medium comprises PPLO broth, phenol red, brilliant blue, L-arginine hydrochloride, yeast extract, ceftriaxone, amoxiclav (sodium salt of amoxicillin/clavulanic acid potassium salt), vancomycin hydrochloride, clarithromycin, fluconazole and distilled water in a predetermined ratio of components.
EFFECT: invention enables to improve the accuracy and to reduce the time of detection of Mycoplasma hominis.
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SUBSTANCE: group of inventions refers to biotechnology. Versions of a nutrient medium for the selective collection of enteric bacteria contain fish-flour pancreatic hydrolysate, or meat peptone, or fish-flour pancreatic hydrolysate and meat peptone (in ratio 1:1), modified purified bile (MPB), sodium chloride, glucose, potassium monophosphate and brilliant green in certain ratio.
EFFECT: inventions enable providing the more effective collection of enteric bacteria, the selective properties of the medium and simplifying the method for producing the nutrient medium.
3 cl, 3 tbl, 9 ex
SUBSTANCE: invention relates to a set of oligonucleotide primers and probes for the identification and subtyping of a bacterial DNA of Pasteurella multocida of serotypes A, B, D, E, F by the PCR method in real time. The invention can be used in genetic engineering and veterinary practice for the detection of a genetic material (DNA) of the bacteria P. multocida of the said serotypes.
EFFECT: improvement of the method.
2 dwg, 5 tbl, 2 ex
SUBSTANCE: probes included in the set have a structure of a "pin" with fluorophore and fluorescence extinguisher at the ends and have complementarity to products of an amplification reaction with primers. The developed set of hybridisation probes can be used for the fluorescent detection of the results of typing the actinobacillus mallei by a method of amplification of differentiating fragments of a genome in the implementation of the epidemiological surveillance of the glanderous infection.
EFFECT: possibility of simultaneous analysis of nine DFR-loci.
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FIELD: food industry.
SUBSTANCE: method involves components selection and preliminary preparation by way of sterilisation with further mixing of the components in equal quantities. The first component of the nutrient medium is a nutrient base containing peptone, sodium chloride, agar and a blue alcohol-based colouring agent, all dissolved in water, at the preset ratio. The second component of the nutrient medium is a fat emulsion sterilised at the preset parameters and consisting of Twin 80 cocoa butter substitute and water mixed in the preset quantities. The nutrient medium and the fat emulsion are mixed after sterilisation either immediately before usage, in equal quantities, with addition of the blue alcohol-based colouring agent in the preset quantity.
EFFECT: invention allows to increase accuracy of lipase-producing microorganisms detection.
SUBSTANCE: electrochemical method for determination of gram-negative pathogenic bacteria in analysed medium suggests using as signal tag of electroactive magnet nanocomposite particles, which are produced before conjugation stage by creation of transition metals or their compounds for electroactive polymer coating on the surface of magnet nanoparticles. Concentration of pathogenic microorganisms is defined by electrochemical response registered directly in result of conversion of electroactive polymer coating.
EFFECT: analysis simplification, increased sensitivity, rapidness, reproducibility, widening the range of electrochemically active tags, using of polymer coating allows reaching high sensitivity and reproducibility of the analysis.
7 dwg, 6 ex
SUBSTANCE: invention relates to medicine and can be used for detection of urease-producing microorganisms, in particular Helicobacter pylori. For this purpose express-analysis is carried out on diagnostic discs to determine urease activity of samples. Discs are made from filter paper, on which urea, acid-base indicator, muromidase and conditioning substances are preliminarily applied. In case if urease activity is present in sample colouration of disc takes place.
EFFECT: invention provides increased sensitivity of method of Helicobacter pylori diagnostics.
SUBSTANCE: invention proposes a method for obtaining a standard specimen of turbidity of bacterial suspensions, a standard specimen of turbidity of bacterial suspensions, use of the standard specimen of turbidity of bacterial suspensions, as well as a set containing the standard specimen of turbidity of bacterial suspensions. The method involves shuttling of chemically neutral borosilicate glass without any carbon dioxide bubbling in presence of thiomersal during 10-70 days. Shuttling is performed in a shaking mode with at least 50-60 oscillations per minute for 5-6 hours a day at oscillation amplitude of at least 5 cm. After shuttling is completed, the obtained suspension is subject to sedimentation for removal of particles with the size of more than 3.5 mcm and less than 0.5 mcm. The standard specimen of turbidity of bacterial suspensions includes a micro suspension of chemically neutral borosilicate glass with the particle size of 0.5-3.5 mcm; with that, content of particles of this fraction is at least 95%.
EFFECT: inventions allow simplifying a method for obtaining a standard specimen of turbidity of bacterial suspensions and simultaneous improvement of stability of the standard specimen.
4 cl, 2 dwg, 2 tbl, 8 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: inventions concern the influenza virus strains A/PR/8/59/M2 (H1N1), A/California/1/66 (H2N2) and A/Tokyo/3/67 (H2N2). The vaccinal strains A/59/M2/California/66/2211 (H2N2) and A/59/M2/Tokyo/67/22111 (H2N2) are reassortants prepared by crossing epidemic viruses A/California/1/66 (H2N2) and A/Tokyo/3/67 (H2N2) with cold-adapted heat-sensitive virus A/PR/8/59/M2 (H1N1). The strains A/59/M2/California/66/2211 (H2N2) and A/59/M2/Tokyo/67/22111 (H2N2) are characterised by heat sensitivity, cold adaption, safety and immunogenicity for laboratory animals. The reassortants has inherited two genes coding surface virus antigens (hemagglutinin and neuraminidase) from the epidemic virus and the rest six genes coding non-glycated proteins, from the attenuation donor A/PR/8/59/M2 (H1N1).
EFFECT: presented inventions can be used in preparing a live influenza intranasal vaccine for adults and children.
3 cl, 2 dwg, 4 tbl, 1 ex