39 results about "Immuno pcr" patented technology

Devices, containers, and methods are provided for performing biological analysis in a closed environment. Illustrative biological analyses include high density nucleic acid amplification and detection and immuno-PCR.

The invention relates to methods and kits for detecting and/or monitoring biological molecules in a test sample. For example, the invention relates to methods and kits for detecting and/or monitoring HIV p24 antigen in human body fluid, biological toxins such as ricin or botulism in an environmental or biological sample, and prion protein from human, deer or bovine, such as PrP^SC, in a biological sample. The antigen detection signal is boosted by amplification of a polynucleotide linked to a detector molecule using methods for nucleic acid amplification technology.

Devices, containers, and methods are provided for performing biological analysis in a closed environment. Illustrative biological analyses include nucleic acid amplification and detection and immuno-PCR.

The present invention relates to a method for the highly sensitive Immuno-PCR detection of an analyte in a sample comprising the use of a nucleic acids containing sample dilution buffer for diluting the sample as well as methods for the preparation of the sample dilution buffer and the use thereof.

This invention relates to a reagent container for detection and/or quantitation of at least one biological or chemical analyte from a sample, said reagent container comprising an inner surface enclosing a volume, wherein volume analytical reactions of at least one analysis for detection and/or quantitation of at least one analyte take place, and at least two reagents of an analysis of an analyte have been dried onto said inner surface and at least a first said reagent has been dried onto a first area of the inner surface distinctly separate, i.e. without any overlap, from a second area of the inner surface onto which at least a second said reagent has been dried. Characteristic for the reagent container is that the first reagent and the second reagent form a pair wherein the first reagent is an enzyme and the second reagent is a substrate of said enzyme, and said pair consists of a nucleic acid polymerase and substrate thereof. This invention also relates to a method for stabilising onto an inner surface of a reagent container dried assay reagents for the detection and/or quantitation of a biological or chemical analyte from a sample, said method comprising the steps of dispensing at least two reagents, a first reagent and a second reagent, needed in the detection of the analyte onto the inner surface of the reagent container; and removing excess water from the reagents, wherein first reagent is dispensed onto a first area of the inner surface distinctly separate, i.e. without any overlap, from a second area of the inner surface onto which the second reagent is dispensed. Characteristic for the method is that the first reagent and the second reagent form a pair wherein the first reagent is an enzyme and the second reagent is a substrate of said enzyme, and said pair consist of a nucleic acid polymerase and substrate thereof. This invention further relates to the use of the reagent container to perform an assay selected from the group consisting of a polymerase chain reaction (PCR) assay, an assay that utilizes a reverse transcriptase, a reverse transriptase polymerase chain reaction, an immuno-PCR assay, a nucleic acid sequence based assay (NASBA), a proximity ligation assay, a ligase chain reaction (LCR) assay, a rolling circle amplification (RCA) assay, and a strand displacement amplification (SDA) assay.

The invention discloses a PCP (aptamer based polymerase chain reaction, A-PCR) based on an aptamer, which relates to a high-sensitivity technology for detecting a micro-target matter. In the technology, the high specificity shown in the integration of a nucleic acid aptamer and a ligand and the high sensibility of the aptamer reacted with a polymerase chain are organically combined; the character of the technology is a method which augments a section of DNA reporter molecule through PCP so as to amplify the combination rate of the aptamer and the ligand, thereby detecting a target ligand. The ligand PCR is one of the most sensitive detecting modes at the moment and can detect a single target molecule in theory. Compared with immuno PCR, the technology has the characteristics of simple operation and process, less affection factor, more reliable result, and the like; the sensibility is improved by 10<2> to 10<8> times than the existing ELISA method; the technology can be broadly applied to the micro detection of protein molecules and small molecules.

The present invention relates to biochemistry, medicine, and molecular biology, in particular to analytical biochemistry and immunochemical assay, and is concerned with a biological microchip for multiple parallel detection and quantitative determination of different compounds. The biochip comprises an array of three-dimensional hydrogel elements which have a predetermined volume, formed on a support by the method of photo- or chemically induced polymerization, and containing biological molecules of the same or different nature (ligands). The invention provides a method for detecting compounds on the biological microchip, comprising identification thereof by mass spectrometry techniques directly on the hydrogel element of the microchip and a method for detecting immunoassay results, consisting in carrying an immuno-PCR and registering the results thereof on the same microchip. The invention also relates to a method of multiple parallel immunoassay of compounds on a biological microchip. Biological microchips for immunoassay and a method of carrying out multiple parallel analysis of compounds can find application in analyzing a wide range of high-molecular and low-molecular compounds, in medicine, pharmacology, food industry, environmental protection, in research work, particularly in proteomics.

The invention belongs to the field of biological technical detection and particularly relates to an immune PCR reagent kit for detecting avian leukemia virus. The reagent kit comprises two antibody oligonucleotide probes, a coupled reaction solution, DNA ligase, protease, a Fast Master mixed solution and a Universal q PRC reaction solution. The immune PCR reagent kit uses a biotin-labeled avian leukemia virus preventing special antibody and a commercialized ortho-position connecting reagent, can detect avian leukemia virus antigen through immune PCR and can quickly detect the avian leukemia virus antigen in a high-throughout mode through the immune PCR without carrying out a complex nucleic acid extracting process on a sample. The immune PCR reagent kit for detecting the avian leukemia virus can be applied to clinical detection and purification of avian leukemia virus and will fill domestic and oversea correlated technique blank.

The invention discloses a construction method of an immunosensor for measuring the DNA (Deoxyribose Nucleic Acid) mark of melamine, belonging to the field of immune PCRs (Polymerase Chain Reactions). The method comprises the following steps of: preparing a DNA-melamine antibody conjugate; coating a PCR tube with a melamine coating antigen; and constructing a melamine immunosensor. The invention provides a DNA-melamine antibody conjugate which is subjected to immunological recognition and fluorescent quantitation PCR amplification of DNA, and an amplified fluorescence signal is used for detecting melamine. The method can be used for detecting melamine at ultrahigh sensitivity, and has the advantages of low detection limit, high detection sensitivity and high specificity; and an effective method is provided for the detection of a trace quantity of melamine medicaments.

The invention relates to a method for jointly characterizing exosome membrane markers and RNA based on aptamer immune PCR. The method mainly comprises the following steps: specifically recognizing andcombining markers on an exosome membrane through aptamers; and simultaneously amplifying the aptamers and the exosome RNA which are combined on the exosome through polymerase chain reaction (PCR), soas to realize the simultaneous characterization of the markers and the RNA on the exosome membrane. The method provided by the invention can realize the simultaneous high-sensitivity detection of themarkers and the RNA on the exosome membrane, saves samples, is simple to operate, and has very high clinical application value.

The invention relates to a detection method of MG7-Ag contained in serum, belonging to the technical field of biology. The detection method comprises the following steps of: accurately quantitating and detecting the content of the trace antigen MG7-Ag contained in the serum by combining a real-time fluorescence quantitative immune PCR detection method on the basis of antigen signal transformation through the identification and the capture of the MG7-Ag by double antibodies; combining a real-time fluorescence quantitative PCR technology with a specific antigen and antibody reaction system to establish a real-time fluorescence quantitative immune PCR technology, wherein the real-time fluorescence quantitative immune PCR technology identifies the MG7-Ag contained in the serum through the specificity of the double antibodies, then respectively combines biotinylated MG7 with biotinylated DNA through streptavidin, converts a content signal of the MG7-Ag contained in the serum into a nucleic acid signal and then enters a real-time fluorescence quantitative PCR expanding way, thereby accurately detecting the trace antigen MG7-Ag contained in the serum.

The invention discloses a biomarker for forecasting the new auxiliary chemotherapy curative effect of breast cancer and a fluorescence quantitative immune PCR kit. The biomarker is at least one of a CCL2 gene, a CCR1 gene, a CXCL10 gene, a CXCL11 gene and an IL2RG gene. The five genes of the biomarker are efficiently expressed in a pCR group; the IL2RG gene has an extremely obvious expression difference (P is less than 0.01) in the pCR group and the NpCR group, and the other four genes have an obvious expression difference (P is less than 0.05) in the pCR group and the NpCR group; the five genes are indicated to be biomarkers for forecasting the new auxiliary chemotherapy curative effect of the breast cancer, and the blindness of new auxiliary chemotherapy is avoided.

Methods and kits related to non-equilibrium, ultrasensitive two-site assays for detecting analytes are provided. In one aspect, a two-site assays for detecting analytes under non-equilibrium analyte binding conditions, using low concentrations of reporter specificity molecule (e.g., reporter antibody) and kits for performing the same is provided. In another aspect, methods for selecting antibodies or specificity molecules with low dissociation constants for use as reporter antibodies in non-equilibrium two-site immunoassays, including two-site immuno-PCR assays, and assays performed with those antibodies, are also provided.

The invention discloses a specific antibody-coated PCR reaction tube and immune PCR detection kit for detecting campylobacter jejuni. A monoclonal antibody which can specifically enrich the campylobacter jejuni in samples pre-coats the PCR reaction tube. The detection kit has the advantages that the detection kit can fast, accurately and sensitively detect the campylobacter jejuni from the samples such as food, the sensitivity of the detection kit can reach 103-104cfu/ml, and the sensitivity of the detection kit is 10-100 times higher than that of conventional PCR; the detection kit organically and integrally combines immunology and molecular biology methods, enrichment and detection of the campylobacter jejuni in the samples can be realized in one PCR tube, and the detection kit is simple to operate, low in cost, fast in detection and accurate in result.

The invention provides a poly-brominated biphenyl homolog semi-antigen and a preparation method thereof. The structural formula of the semi-antigen is represented in the description. The preparation method comprises a step of reacting TBBPA with bromo-acetic acid in an alkaline environment to introduce a carboxyl group into the terminal end of the phenolic hydroxyl group of TBBPA so as to obtain the semi-antigen. The provided semi-antigen preparation method has the advantages of simpleness, high stability, low cost, and easiness for industrial production. TBBPA semi-antigen can be coupled to carrier protein to prepare TBBPA artificial holoantigen through a carbodiimide method, a mixed acid anhydride method, or an activated ester method. The holoantigen can be used to prepare antibody having the advantages of high specificity and high titer and establish an immuno-PCR biological bar-code method.

The invention belongs to the field of measurement of plant hormones, and provides a measuring method of immune PCR of abscisic acid. The method comprises the steps of usingglutaraldehyde to pretreat the inner surface of a PCR tube/ plate, using a single anti-abscisic-acid antibody to cover the surface of the treated PCR tube/ plate to synthesize a probe mixture, adding the probe mixture into the treated PCR tube/ plate, then adding an abscisic acid sample to be measured into the PCR tube/ plate to be reacted, conducting amplification in real-time PCR equipment, and according to the amplification result, determining the content ofabscisic acid. A comparison experiment shows that by utilizing glutaraldehyde to pretreat the inner surface of the PCR tube/ plate, the antibody covering capability can be improved by 20-30%; the three substances of a biotinylation ABA polyclonal antibody, biotinylation DNA and avidin are further utilized to synthesize the probe mixture, the repeatability of ABA measurement is improved, the detection lower limit of the immune PCR method can reach 10 ng/L, close to the detection result of liquidchromatography mass, and the problem that organism in-vitro plant hormones aredifficult to directly react with biomacromolecules is solved.

The present invention belongs to the field of immunological detection methods. A purpose of the present invention is to provide a detection kit and a detection method using antibody to modify an immuno-PCR reaction. The detection method comprises synthesis of DIBO-labeled ssDNA, biotin modification of Insulin monoclonal antibody, preparation of ssDNA-modified immuno-PCR micro-pore plate of Insulin monoclonal antibody, establishment of Immuno-PCR standard curve of Insulin, and other steps. According to the present invention, with the application of the antibody modification technology in the immuno-PCR reaction, the detection on the ultra low concentration biotin marker can be performed, the sensitivity of the detection method of the present invention can exceed 1000 times the sensitivity of the conventional ELISA method, and the extremely-low concentration biomarker can be detected.

The invention discloses a specific antibody coated PCR (polymerase chain reaction) tube and an immune PCR detection kit which are used for detecting Listeria monocytogenes. The PCR reaction tube is precoated with monoclonal antibodies which can specifically enrich Listeria monocytogenes in samples. The detection kit can quickly, accurately and sensitively detect the Listeria monocytogenes from the samples like food, and the sensitivity can be 10<3>-10<4>cfu/ml and is increased by 10-100 times compared with the sensitivity of the conventional PCR. The PCR reaction tube and the immune PCR detection kit have the advantages that immunology and a molecular biological method are organically combined into a whole, enrichment and detection of Listeria monocytogenes in the samples can be realized in one PCR tube, operation is easy, cost is low, detection is quick and results are accurate.

The invention discloses a PCR (polymerase chain reaction) tube coated by a specific antibody and an immune PCR (polymerase chain reaction) detection kit, which are used for detecting staphylococcus aureu. A monoclonal antibody capable of specifically enriching staphylococcus aureu in a sample is pre-coated on the PCR tube, the detection kit disclosed by the invention is capable of fast, accurately and sensitively detecting staphylococcus aureu from samples such as food, the sensitivity can achieve 10<3>-10<4>cfu/ml, and is improved by 10-100 times in comparison with that of the conventional PCR. The immunology and a molecular biology method are organically combined to enrich and detect the staphylococcus aureu in the sample in the PCR tube, the operation is simple, the cost is low, the detection is fast and the result is accurate.