SUBSTANCE: invention relates to biochemistry. Method of determining sensitivity of tumor cell growth to inhibition by EGFR kinase inhibitor is disclosed, which involves determining ERBB2 gene methylation status in tumor cell sample, reduced level of methylation ERBB2 gene means, that less than 50 % of possible methylation sites in part of ERBB2 gene are methylated, and indicates that growth of tumor cells is sensitive to inhibition by inhibitor EGFR. Method of detecting patient with malignant tumor is also described, favorable effect on which can have treatment with help of EGFR inhibitor. Method of selecting therapy for patient with malignant tumor is described. Invention extends range of methods for prediction of response to anticancer therapy.
EFFECT: method of determining ERBB2 gene overexpression in cell is disclosed.
31 cl, 8 dwg, 6 ex
SUBSTANCE: method includes treating a sample with a denaturing buffer solution, sorbing short RNA on a fibre glass sorbent in the presence of a chaotropic agent, followed by washing off non-bonded biopolymers and chemical reagents from the sorbent and eluting the end product. The biological fluid sample undergoes two-step denaturing, wherein at the first step two volumes of the denaturing buffer solution are added to the sample and at the second step two volumes of 96% ethanol and equal volume of chloroform are added to the obtained mixture, followed by stirring and separating the residue by centrifuging. Non-bonded biopolymers are washed off from the sorbent twice with the buffer solution and elution of the end product from the sorbent is carried out with the buffer solution.
EFFECT: simple method, short duration of the method and high output of short RNA.
3 cl, 3 tbl, 4 ex
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
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
FIELD: medicine, psychiatry.
SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.
EFFECT: more objective prediction of disease development.
FIELD: biotechnology, medicine, proteins.
SUBSTANCE: invention describes new polypeptide in isolated form relating to subfamily of superfamily human immunoglobulins (Ig-Sf). This polypeptide shows at least 70% of homology level with amino acid sequence of murine molecules CRAM-1 or CRAM-2 regulated by the confluence of adhesive (figures 3, 6 are represented in the claim). Also, invention relates to antibodies showing specificity with respect to the polypeptide. Antibodies and soluble polypeptide can be used for treatment of inflammation and tumors. Invention describes polynucleotide or oligonucleotide encoding the full-size polypeptide or its moiety and represents primer, probe, anti-sense RNA and shows the nucleotide sequence that is identical conceptually with human CRAM-1. Invention provides preparing new adhesive proteins from superfamily Ig-Sf that are regulated at the transcription level in endothelium by effect of tumors. Invention can be used for treatment of different diseases, in particular, inflammatory responses.
EFFECT: valuable medicinal properties of polypeptide.
19 cl, 33 dwg, 1 ex
FIELD: biotechnology, genetics.
SUBSTANCE: invention relates to methods used for detecting low frequency mutations occurrence in gene encoding cytochrome b. Method involves isolation of DNA from known fungi for constructing oligonucleotide probe or primer. Then polymerase chain reaction (PCR) is carried out for assay of binding the nucleotide probe with amplicon generated by this reaction, or the presence of amplicon is detected that is generated as result of PCR using indicated primers. Invention provides rapid and precise detection of mutations conferring resistance of fungus against fungicide.
EFFECT: improved diagnostic methods for detecting mutations.
24 cl, 18 dwg, 14 tbl, 18 ex
FIELD: genetic engineering, molecular biology, pharmacy.
SUBSTANCE: invention relates to methods of genome screening and can be used for identification of pharmacological agent in vegetable extract. Method is realized by treatment of cells with a vegetable extract, isolation of protein or RNA from these cells, identification of isolated protein or RNA wherein their concentration differs from that in untreated cells and detection of compound(s) in indicated vegetable extracts. Then cells are treated with the found compound(s), protein or RNA are isolated from cells treated with this compound(s) and compound(s) are identified that cause the stimulation or inhibition of expression of protein or RNA wherein their concentration differs from that in untreated cells. Invention provides carrying out the characterization of biological properties of vegetable extract and to detect the individual compound(s) that elicit unknown or disclosed biological property of this extract.
EFFECT: improved identifying method.
FIELD: medicine, molecular biology, polypeptides.
SUBSTANCE: invention describes homogenous polypeptide ligand mpI representing polypeptide fragment of the formula: X-hTPO-Y wherein hTPO has amino acid sequence of human fragments TPO (hML); X means a amino-terminal amino-group or amino acid(s) residue(s); Y means carboxy-terminal carboxy-group or amino acid(s) residue(s), or chimeric polypeptide, or polypeptide fragment comprising N-terminal residues of amino acid sequence hML. Also, invention relates to nucleic acid encoding polypeptide and expressing vector comprising nucleic acid. Invention describes methods for preparing the polypeptide using cell-host transformed with vector, and antibodies raised against to polypeptide. Invention describes methods and agents using active agents of this invention. The polypeptide ligand mpI effects on replication, differentiation or maturation of blood cells being especially on megacaryocytes and progenitor megacaryocyte cells that allows using polypeptides for treatment of thrombocytopenia.
EFFECT: valuable medicinal properties of polypeptide.
21 cl, 92 dwg, 14 tbl, 24 ex
FIELD: medicine, biology, molecular biology.
SUBSTANCE: invention proposes a new method for differential diagnosis of representatives of family Chlamydiaceae. Method involves isolation of DNA of pathogen, amplification using real-time polymerase chain reaction (PCR) and primers CM1 and CM2 exhibiting specificity to 5'-terminal fragment of gene omp1 followed by post-amplification analysis of curves of PCR-products melting in the presence of nonspecific fluorescent dye SYBR Green I for separation of Chlamydia species and electrophoretic separation of PCR-products. Identification of species is carried out on the basis of differences in PCR-products melting point wherein melting point curves of all fragments of omp1 are characterized by the presence of two peaks reflecting two-stage dissociation of DNA chains in sites with different A/T-saturation degree. Proposed method provides carrying out the differentiation of all species of Chlamydia that are pathogenic for humans. Except for, method provides carrying out the differentiation of Chlamydiaceae causing diseases in animals and also method is simple, rapid and can be used for direct diagnosis of clinical material samples. Invention can be used in medicine, veterinary science and virology for differential diagnosis of representatives of family Chlamydia.
EFFECT: improved method for diagnosis.
FIELD: medicine, biology, molecular biology.
SUBSTANCE: invention proposes a new method for differential diagnosis of representatives of family Chlamydiaceae. Method involves isolation of DNA from pathogen, amplification of target using primers CM1 and CM2 showing specificity to 5'-terminal fragment of gene omp1 and electrophoretic separation of polymerase chain reaction (PCR) products. Electrophoresis is carried out in agarose gel with addition of sequence-specific DNA-ligand - bis-benzimide-PEG. The species belonging of PCR-products is determined by comparison of the migration rate of PCR-products in gel with electrophoretic mobility of control. Proposed method provides carrying out the differentiation of all species of family Chlamydiaceae and method is simple and rapid and can be used for direct diagnosis of clinical material samples also. Invention can be used in medicine and virology for differential diagnosis of representatives of family Chlamydiaceae.
EFFECT: improved method for diagnosis.
2 dwg, 2 ex
FIELD: molecular biology, medicine, biochemistry.
SUBSTANCE: invention proposes a method for assay of mononucleotide changes in the known sequences of nucleic acids. Method involves hybridization with PCR-amplified matrix DNA and the following ligation a tandem on its consisting of tetranucleotide that comprises the diagnosed change and two oligonucleotides of the size 8-10 nucleotides being one of that is immobilized on surface of a solid-phase carrier through a 5'-phosphate linker, and the second oligonucleotide is labeled by 3'-end with biotin label. Then tetranucleotide is hybridized with the matrix DNA chain between two oligonucleotides directly that can be ligated with immobilized oligonucleotide through the 5'-end and with non-immobilized oligonucleotide through the 3'-end. The ligation product is detected by its transformation to enzyme label through the complex with high-affinity enzymatic catalyst followed by development of enzyme label in the presence of chromogenic, luminogenic or fluorogenic substrates. Applying a method provides preparing the simple and highly selective agent used for detection of known changes in gene structure.
EFFECT: improved assay method.
8 cl, 3 dwg, 30 ex
FIELD: genetic engineering, biotechnology, biochemistry, agriculture, food industry, medicine.
SUBSTANCE: invention relates to the transformation of plant with nucleic acid encoding enzyme Δ6-desaturase in C. elegans that results to preparing a plant with enhanced content of gamma-linolenic acid and resistance to cold. Desaturase extracted from the plant can be used for preparing a drug used for treatment of disorder in body associated with deficiency of gamma-linolenic acid in it.
EFFECT: valuable biological properties of genes and desaturases.
36 cl, 9 dwg, 2 ex
FIELD: medicine, hematology.
SUBSTANCE: one should isolate DNA out of peripheral blood lymphocytes due to polymerase chain reaction (PCR) technique of DNA synthesis, carry out genotyping for polymorphism of promoter area of TNF-alpha and TNF-beta gene. While detecting genotype LT*22 being characterized by availability of gene TNF-beta mutation in homozygous state, detect persons predisposed to the development of chronic lympholeukosis, and at certain combinations of genotypes TNF*22/LT*22 or TNF*12/LT*11 in patients with chronic lympholeukosis on should predict an aggressive flow of this disease.
EFFECT: higher accuracy of prediction.
3 ex, 3 tbl