Method of detecting of cell culture contamination with viruses by immunoperoxidase method

FIELD: biochemistry.

SUBSTANCE: invention relates to biochemistry. Method of detecting contamination with viruses of cell culture by immunoperoxidase method is described, involving reaction of antigen with labelled antibodies in clean cell culture and accounting of reaction results by color density of formed complex under microscope. Wherein reaction uses tablets with analyzed cell culture, contaminated with diarrhoea virus, and after adding into tablet wells of 0.10–0.12 ml of specific homologous serum diluted by 1:64 – 1:128 it is incubated, washed, then 0.10–0.12 ml of anti-specific immunoenzymometric conjugate is introduced, consisting of peroxidase marked antibodies against globulins of cattle blood serum, it is incubated, washed, substrate mixture is introduced consisting of 5-aminosalicylic acid and hydrogen peroxide, and 30–40 minutes later results are accounted under light microscope by formation of brown color in virus-containing cells. Invention improves control system of cell lines latent contamination with viruses used in laboratory practice and biological industry.

EFFECT: invention allows to prevent biological contamination and is extremely important stage in production of vaccines and diagnostic preparations; this method can be used for certification at absence of viruses in again coming cell cultures; invention is practically feasible, cheap method of monitoring of cell cultures for content of virus and may be recommended for extensive use both in scientific research and practical virological laboratories, and in production in making diagnostic preparations and vaccines.

1 cl, 2 ex

 



 

Same patents:

FIELD: biotechnology.

SUBSTANCE: strain of bacteria Rhodococcus sp. LER-12 having the ability to dispose quickly of crude oil and petroleum products (diesel fuel, motor oil, hydraulic oil, gas condensate) is deposited in the RNCM under the registration number Rhodococcus sp. RNCM Ac-2626D and can be used for purification of soils and water from contaminations with crude oil and petroleum products, in a wide temperature range from +4 to +30°C.

EFFECT: invention enables to improve the quality of purification of soil and water from crude oil and petroleum products.

3 tbl, 5 ex

FIELD: biotechnology.

SUBSTANCE: strain is deposited in the State collection of pathogenic microorganisms "SCIM-Obolensky" under the number F-11. The culture fluid centrate obtained as a result of culturing of said the strain on the liquid nutrient medium has high activity against anthrax causative agent. The zone of inhibition of growth of Bacillus anthracis is 20-25 mm and 35-45 mm when the culture fluid centrate of the said strain is used in an amount of 10 mcl and 100 mcl, respectively.

EFFECT: strain of micromycete Trichoderma hamatum has antibacterial activity against anthrax causative agent Bacillus anthracis.

4 tbl, 3 ex

FIELD: biotechnology.

SUBSTANCE: nutrient medium comprises sodium chloride, potassium chloride, magnesium sulphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, calcium chloride, sodium bicarbonate, L-arginine, L-glutamine, L-tyrosine, L-tryptophan, calcium pantothenate, pyridoxal HCl, thiamine, meso-inositol, nicotinamide, riboflavin, folic acid, glucose, enzymatic hydrolyzate of muscle proteins, lactalbumin enzymatic hydrolyzate, sodium salt of benzylpenicillin with activity of 0.09×105-1.1×105, kanamycin sulphate with activity of 0.09×105-1.1×105, nystatin with activity of 6×104 to 6.5×104 IU/l, choline chloride, succinic acid and distilled water in a predetermined ratio of components.

EFFECT: invention enables to improve the quality of the product obtained.

1 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. The strain of fungus Aspergillus niger B-6 is the producer of citric acid. The strain Aspergillus niger is deposited in the joint Russian Collection of Agrocultural Microorganisms RAAS (RCAM) under registration number RCAM02149 and can be applied in the production of citric acid.

EFFECT: invention makes it possible to increase the output of citric acid.

2 tbl, 6 ex

FIELD: biotechnologies.

SUBSTANCE: method of production of nanodisperse additive for concrete is offered. Cyanobacteria of the sort Pseudanabaena sp. 0411 or Leptolyngbya laminosa 0412 are cultivated in a nutrient medium at the temperature 23-25°C. The nutrient medium is Z-8 medium with adding of sodium silicate solution neutralized by 2 M HCl. Ratio of solution of silicate of sodium and Z-8 5:1 medium. Cultivation is carried out in the bioreactor at continuous lighting and mixing during 10 days with the subsequent removal of nutrient medium residue. Then the culture is poured by 30% hydrogen peroxide solution and heated up to 70°C. The produced biosilicated nanotubes are washed out by distilled water with subsequent treatment in a mechanical activator in the water medium of anion surface-active substance of naphthalene formaldehyde type at the ultrasound frequency 35 kHz and concentration of solid phase 5% up to the particle size 85-250 nanometres.

EFFECT: increase of mobility of concrete mix, curing acceleration, increase of strength and density of concrete, decrease of water absorption.

3 tbl, 1 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanovermiculite and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of nitrogen-fixing and phosphate-mobilising microorganisms.

1 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanobentonite and distilled water.

EFFECT: enhanced growth rate of phosphate-mobilising and nitrogen-fixing microorganisms.

1 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nano sapropel and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of phosphate-mobilising and nitrogen-fixing microorganisms.

1 tbl, 15 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanozeolite and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of nitrogen-fixing and phosphate-mobilising microorganisms.

1 tbl, 15 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology and concerns the strain Escherichia coli BL21(DE3)Gold/pETmin-CypA producing human recombinant cyclophilin A. The characterised strain is produced by cell transformation of the strain BL21(DE3)Gold by pETmin-CypA plasmid. The plasmid has a size of 5865 base pairs and contains XhoI-NdeI fragment of pET-22b(+) vector carrying ampR ampicillin resistance gene, a promoter and RNA-polymerase T7 phage terminator and a polylinker. A fragment produced by PCR with using the oligonucleotide primers 5'-TTATACATATGGTCAACCCGACCGTGTTCTTC and 5'-TTTCTCGAGTTATTCGAGTTGTCCACAGTCAGC of pCyPAwt/pGEX-2TK plasmid with the closed fragment of hrCpA (human recombinant cyclophilin A) gene having a size of 498 base pairs is cloned at XhoI-NdeI sites.

EFFECT: presented strain is high-producing; it requires no complicated clean-up system and can be used in cancer treatment.

2 dwg

FIELD: medicine.

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

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

FIELD: biotechnology.

SUBSTANCE: proposed primers comprise endonuclease cleavage sites, flanking genomic regions encoding the glycoproteins Gn and Gc, and the nucleoprotein N, for obtaining libraries of genes encoding glycoproteins Gn and Gc and N nucleoprotein of Rift Valley fever virus.

EFFECT: invention can be used in creating a bank of nucleotide sequences encoding immunodominant proteins of Rift Valley fever virus Gn, Gc and N, which can be used for creation of diagnostic and vaccine preparations based on recombinant technologies.

3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the field of biotechnology and can be used for the identification of heteromultimeric ubiquitins possessing an ability to bind with a ligand-antigen. The method includes the contact of a totality of heterodimeric modified ubiquitins, including two ubiquitin monomers, bound to each other by a head-to-tail scheme, with the potential ligand in a display way. Each of the said monomers is modified in a different way and contains 5-8 substitutions in positions 2, 4, 6, 8, 62, 63, 64, 65, 66 and 68 SEQ ID NO:1. After that, a heterodimeric modified protein, which has bound with the ligand with the binding affinity Kd in the range of 10-7-10-12 M and the monovalent binding activity. Claimed are DNA-libraries, responsible for obtaining a population of the said heteromultimeric ubiquitins, as well as libraries of proteins, obtained by the expression of the said DNA-libraries.

EFFECT: invention makes it possible to obtain the novel bonding proteins based on heteromultimeric ubiquitin, capable of specific high affinity binding with the selected ligands.

8 cl, 17 dwg, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: there are presented a composition and a method for producing it. The characterised composition contains: an effective amount of a viral antigen, which represents a live attenuated rotavirus pre-processed in 0.1% human serum albumin, and a pharmaceutically acceptable buffer. A method for producing a composition involves growing Vero cell culture pre-cultured in the presence of 5% foetal calf serum and 0.1% human serum albumin, infecting the above Vero cell culture with the live attenuated rotavirus, propagating the virus in the cell culture and adding a pharmaceutically acceptable buffer to the above virus.

EFFECT: presented inventions can be used to prevent rotavirus infection and/or rotavirus gastroenteritis.

11 cl, 17 dwg, 4 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: inventions deal with infectious molecule of nucleic acid, coding infectious porcine Torque teNO viruses (PTTV), which contains at least one copy of genome sequence, selected from the group, consisting of sequences, corresponding to genotypes or subtypesPTTV1a-VA, PTTV1b-VA, PTTV2b-VA and PTTV2c-VA, as well as to biologically functional plasmid or viral vector, containing such infectious nucleic genome sequence, and host-cell, containing such plasmid or vector. In addition claimed inventions include live, attenuated expressible with vector application and purified recombinant capsid subunit or killed viral vaccines for protection against PTTV infection, as well as methods of immunisation of pigs against PTTV viral infection by said vaccine introduction.

EFFECT: characterised inventions can be used to prevent infection, caused by porcine Torque teNo virus.

23 cl, 53 dwg, 5 tbl, 24 ex

FIELD: biotechnology.

SUBSTANCE: characterised strain was isolated from diseased pigs and produced by serial passages on sensitive hetero- and homologous cell cultures and deposited in the collection of the FSBI "Federal Animal healthcare centre" under the registration number of FMD virus strain A No.2187/Kuti/2013 (production). The presented strain is reproduced in monolayer cell culture of porcine kidney (PK), passaged cell cultures of kidney of Siberian mountain ibex (SMIK-30), VPK-21 and IB-RS-2. During 18÷24 hours of incubation the virus yield in the said cell cultures reaches the values of 6.0÷7.0 lg TCD50/cm3. At high multiplicity of infection (1÷10 TCD/cell) causes TCID50 after 5 hours, while maintaining the original characteristics when passaging in cell cultures for 5 passages.

EFFECT: invention can be used to monitor the antigenic and immunogenic activity and for producing biological products for diagnostics and specific prophylaxis of FMD of type A.

6 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to oncology. The subject of the invention is a new strain of the Sendai virus Sen293nsk1 adapted to effective replication in the human cell culture HEK293. The produced strain of the Sendai virus possesses lower virulence for laboratory mice and higher ability to destroy human tumour cells. The strain is supposed to be used experimentally as a therapeutic oncolytic preparation for treating malignant diseases. The invention can be used in treating oncologic diseases.

EFFECT: invention enables providing higher clinical effectiveness in oncologic diseases by using the murine Sendai virus non-pathogenic for humans, possessing an increased oncolytic activity and intensifying anti-tumour immunity.

1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to such compositions and pharmaceutical compositions, which include poxviruses, and namely to those, which include extracellular enveloped viruses. Claimed invention also relates to such method, which is intended for production of poxviruses, as well as poxviruses, obtained in accordance with claimed invention. In addition, claimed invention also relates to application of claimed poxviruses and said composition for medication preparation.

EFFECT: obtaining pharmaceutical compositions, which include poxviruses.

11 cl, 3 dwg

FIELD: biotechnology.

SUBSTANCE: method of detection and differentiation of a genome of the vaccine strain B-82 of the myxoma virus of rabbits from the field isolates comprises: extraction of the DNA from the biological material, posing a multiplex polymerase chain reaction using synthesised primers complementary to regions of genes M130R and M151R of the myxoma virus of rabbit, and having the following nucleotide composition: MF 5'TGG-AGC-TTT-TCA-AGC-ATT 3', MR 5'ATA-TCT-CGG-CTC-TAG-GGC-GAG 3', MZ 5' [FAM]-AG-CGT-CGG-ACG-TCT-TCG-TT-[RTQ1] 3', VF 5'AGC-CCT-ATA-AAC-CCG-TAG-ACG-AAC 3', VR 5'CAA-GCT-TTT-TTT-TAT-CCT-CGT-CCG 3', VZ 5' [R6G]-TCG-ACG-GTT-TCG-TCC-GCC-TTC-TTG-[BHQ2] 3', DNA amplification of the virus and evaluation of the reaction. The present inventions may be used in veterinary virology for the detection and differentiation of the vaccine strain B-82 of the myxoma virus of rabbits from the field isolates.

EFFECT: increase in accuracy.

2 cl, 6 tbl, 6 ex

FIELD: biotechnology.

SUBSTANCE: method comprises applying to fermented milk products by mixing of liquid culture of strain of propionic acid bacteria Propionibacterium freudenreichii R13-slg RNCIM B-11325 in the amount of 0.5-2.5 vol. % or freeze-dehydrated powder of this culture in an amount of 0.04-0.10 wt %, after which the product storage at 0-4°C for up to 2-3 months, at 20-22°C for up to 30-35 days and at 30-35°C for up to 2-8 days.

EFFECT: enrichment of fermented milk products with viable probiotic cells and vitamin B12 and safety of fermented milk products, which is recorded by the absence of fungal growth, the absence of change of taste, smell, formation of bubbles.

2 dwg, 1 tbl, 23 ex

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