1225results about How to "Strong specificity" patented technology

The invention provides a Chinese cabbage clubroot susceptibility related gene BrSWEET14a, a construction method and application, belongs to the technical field of plant genetic engineering, and particularly relates to a Chinese cabbage clubroot susceptibility related gene BrSWEET14a. The CDS sequence of the Chinese cabbage BrSWEET14a is as shown in SEQ ID No. 1 (sequence identifier number 1). The gene is transformed into Col-type arabidopsis thaliana through an agrobacterium flower immersion transformation method, a BrSWEET14a heterologous expression arabidopsis thaliana strain is obtained, and the result shows that T-DNA insertion mutant of the BrSWEET14a homologous gene Atsweet14 is more resistant to clubroot than that of the Col-type arabidopsis thaliana, and heterologous expression of Chinese cabbage BrSWEET14a leads to reduction of disease resistance of arabidopsis thaliana to plasmodiophora brassicae. Therefore, the Chinese cabbage BrSWEET14a is closely related to the clubroot disease, and the gene and the homologous gene of the gene in other cruciferous plants can be applied to breeding of the cruciferous plants and other horticultural plants and have a good application prospect.

The invention discloses an anti-human hepatitis B e antigen monoclonal antibody and application thereof, and relates to the technical field of hepatitis B detection. In a heavy chain variable region of the monoclonal antibody, an amino acid sequence of a complementarity determining region CDR1-3 is as shown in SEQ ID NO.1-3 or has at least 95% of homology with the sequence as shown in SEQ ID NO.1-3; in the light chain variable region, amino acid sequences of complementary determining regions CDR1 and CDR3 are shown as SEQ ID NO.4 and SEQ ID NO.5 or have at least 95% of homology with the sequences shown as SEQ ID NO.4 and SEQ ID NO.5, and an amino acid sequence of a complementary determining region CDR2 is FAS. The monoclonal antibody provided by the invention can well recognize a natural hepatitis B e antigen, and the titer of a supernatant cultured by a B cell can reach 1: 10000, so that the monoclonal antibody can be completely applied to detection research of the human hepatitis B e antigen. The chemiluminescence reagent prepared by using the monoclonal antibody has the advantages of high specificity, strong anti-interference capability, high detection sensitivity, good stability and the like, almost has no leak detection, and can replace an imported reagent.

The invention provides an electrochemical biosensor based on a nanoparticle modified multilayer porous membrane and application of the electrochemical biosensor. The nano particle modified multilayer porous membrane is composed of a plurality of modified nano particles among membranes with nano pore channels. The electrochemical biosensor constructed based on the nanoparticle modified multilayer porous membrane has the following advantages: (1) good conductivity and stability; and (2) multiple capture probes are modified on the surfaces of the nano pore channels of the multi-layer porous membrane, so that multiple different targets can be respectively detected, and multiple different targets can also be detected at the same time. And (3) enzyme-free double-target sensitive normal-temperature amplification-free detection can be carried out. (4) preparation steps are simple, consumed time is short, and operability is high; and (5) the sensitivity and the specificity are high.

The invention discloses a camel source single-domain antibody GH12 for recognizing a conformation B cell epitope in clostridium difficile glutamate dehydrogenase and application of the camel source single-domain antibody GH12. The nucleotide sequence of the camel source single-domain antibody GH12 is shown as SEQ ID NO: 1, and the amino acid sequence of the camel source single-domain antibody GH12 is shown as SEQ ID NO: 2. The camel source single-domain antibody GH12 disclosed by the invention has the advantages of strong specificity, high sensitivity, good accuracy and the like, and it is proved that the camel source single-domain antibody GH12 can be used for detecting recombinant expressed CD-GDH and endogenous CD-GDH by enzyme-linked immunosorbent assay (ELISA), protein dot blot hybridization (DB) and other methods.

The invention discloses a production method of distiller's grain fermentation liquid fertilizer specially applied to roxburgh rose. The distiller's grain fermentation liquid fertilizer is prepared bythe step of evenly mixing 50 to 80 parts of distiller's grain fermentation extracting solution, 0.5 to 1 part of microelement, 5 to 10 parts of seaweed meal, 5 to 10 parts of natural active organism,2 to 5 parts of potassium bicarbonate, 2 to 5 parts of natural active organism and 2 to 5 parts of natural surface active agent. A preparation method of the distiller's grain fermentation extracting solution comprises the steps: evenly stirring 50 to 80 parts of distiller's grain, 50 to 150 parts of rice washing water, 1 to 2 parts of EM microflora fermentation liquid and 0.5 to 1 part of sugar, performing composting fermentation at the room temperature to obtain a distiller's grain fermentation material, stirring and extracting through a sodium carbonate water solution, filtering and performing evaporation and condensation on obtained filtrate until the density is 1.1 to 1.2 to obtain distiller's grain fermentation extracting solution. The distiller's grain fermentation liquid fertilizerdisclosed by the invention has a good use effect, meanwhile can effectively prevent pest and disease damage to the roxburgh rose and is high-efficiency environment-friendly liquid fertilizer.

The invention provides a fucosyl transferase polypeptide, which can be applied to the synthesis of human milk oligosaccharides (HMOs), and belongs to the technical field of enzyme engineering. The fucosyltransferase polypeptide provided by the invention comprises polypeptides with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% same amino acid sequences as polypeptides shown in amino acid sequences such as SEQ ID NO: 1-25. According to the transferase polypeptide disclosed by the invention, the specificity of a polypeptide catalytic substrate is also improved while the ability of the polypeptide as shown in SEQ ID NO: 1 to catalyze and synthesize 2 '-FL is improved.

The invention discloses a primer pair and a method for identifying a new Oudemansiella oosporoides strain HCS, and belongs to the technical field of specific molecular markers, the preservation number of the new Oudemansiella oosporoides strain HCS is CGMCC NO.41593, and the sequences of the primer pair Y6-F1 / Y6-R2 are as shown in SEQ ID NO.2 and SEQ ID NO.3. The specific method comprises the following steps: extracting DNA of an Oudemansiella oosporoides strain to be detected; carrying out PCR (Polymerase Chain Reaction) amplification by taking the extracted DNA of the sample to be detected as an amplification template and a molecular specific labeled primer consisting of the sequence shown by the primer as an amplification primer; and carrying out agarose gel electrophoresis detection on the PCR product. The primer pair has the advantages that the primer pair can be used for quickly identifying the strain HCS, the operation is simple and convenient, the specificity is strong, the accuracy is high, and a technical basis is provided for protecting a new variety of the strain HCS.

The invention discloses preparation of an antibody capable of detecting multi-subtype KPC and application of a kit of the antibody, and belongs to the field of biological medicine. According to the kit for detecting the common subtypes of KPC and the application of the kit, the mouse antibodies are prepared through immunization of conserved antigen epitopes of all the subtypes of KPC, in-vitro screening and pairing are carried out in multiple modes, then an antibody pair which is good in specificity and high in affinity and can be combined with multiple subtypes is obtained, the titer of the two antibodies can reach one million or above, and the kit is high in specificity, good in affinity and capable of being used for detecting the common subtypes of KPC. Amino acid sequences of a heavy chain variable region and a light chain variable region are obtained through sequencing, and rapid and stable expression can be realized after recombination. The product for detecting the carbapenem KPC is developed on the basis of an immunochromatography platform, and the kit has the advantages of high sensitivity, good specificity, good stability, capability of covering most epidemic KPC subtypes and rapidness and convenience in detection, and provides important help for clinical auxiliary diagnosis and medication guidance.

The invention belongs to the field of biomedicine, and particularly relates to application of ABCG1 in diagnosis or auxiliary diagnosis of HIV (human immunodeficiency virus) combined hyperlipidemia. Through detection, the expression of the ABCG1 in the blood of a patient with HIV combined with hyperlipidemia is obviously reduced, the ABCG1 is verified to have the characteristics of high sensitivity and high specificity, whether hyperlipidemia exists in an HIV infected group or not can be diagnosed or diagnosed in an auxiliary manner by detecting the expression of the ABCG1, guidance is provided for early intervention of hyperlipidemia in the HIV infected group, and the method is suitable for clinical application. Wide clinical application prospects are realized.

The invention relates to the technical field of molecular biological detection, and discloses a composition for detecting pathogenic microorganisms, a kit and application of the composition and the kit. The composition provided by the invention can simultaneously comprise specific primers aiming at a plurality of different pathogenic microorganisms and has excellent specificity and detection sensitivity to the specific pathogenic microorganisms, and different primers do not generate dimerization, so that the different pathogenic microorganisms can be simultaneously, rapidly and accurately detected.

The invention discloses a chromogenic culture medium for detecting gram-negative bacteria in bovine mastitis and a test plate thereof, the culture medium comprises a nutritional ingredient mixture, a composite inhibitor, a composite chromogenic enzyme substrate and cold water gel, the compound chromogenic enzyme substrate is red tetrazole, 5-bromo-6-chloro-3-indole-beta-D-galactoside, and 5-bromo-4-chloro-3-indole-beta-D-glucopyranoside, and the compound chromogenic enzyme substrate is a compound chromogenic enzyme substrate, and the compound chromogenic enzyme substrate is 5-bromo-6-chloro-3-indole-beta-D-glucopyranoside. The chromogenic culture medium and the test board prepared from the chromogenic culture medium can effectively inhibit the growth of non-gram negative bacteria, have the characteristics of strong specificity, high sensitivity, short detection period, easy observation and low detection cost, and have huge development potential and wide application prospects in microbiological detection.

The invention relates to the field of biological medicine, in particular to a nucleoside analogue and application thereof to reduction of off-target activity of oligonucleotide drugs. The nucleoside analogue provided by the invention can be used for preparing oligonucleotide drugs, and can be coupled to a seed region of siRNA, so that the specificity of siRNA is efficiently improved, and the silence effect of a target gene can be maintained while the off-target effect of siRNA is reduced.

The invention belongs to the technical field of medicine detection, and particularly relates to a high-sensitivity liquid chromatography method for detecting mutagenic impurities in atomoxetine hydrochloride key starting material 3-chlorobenzene propanol, the method adopts an HPLC (High Performance Liquid Chromatography) analysis method, a chromatographic column adopts octadecylsilane chemically bonded silica as a filler, and the composition of a mobile phase A is a buffer solution; the mobile phase B is methanol or acetonitrile; carrying out gradient elution on the mobile phase; the flow velocity is 0.4 to 0.8 ml / min; the detection wavelength is 230 to 250 nm; by adopting an HPLC analysis method, the content of mutagenic impurities in the key starting material 3-chlorobenzene propanol of atomoxetine hydrochloride can be effectively separated and measured, and the method is high in sensitivity, strong in specificity and good in accuracy and repeatability, and has extremely important significance on quality control of the key starting material 3-chlorobenzene propanol of atomoxetine hydrochloride.

The invention provides a primer probe group for simultaneously detecting American strains and European strains of porcine reproductive and respiratory syndrome viruses, a detection method and application, and particularly belongs to the technical field of porcine pathogen detection products. The invention provides a primer probe group for simultaneously detecting American strains and European strains of porcine reproductive and respiratory syndrome viruses. The primer probe group comprises universal forward primers PRRSV1-F and PRRSV2-F for detection, reverse primers PRRSV1-R and PRRSV2-R for detection and a universal probe PRRSV-P. A target gene detected by the primer probe group is a 3'non-coding region gene (3 'UTR) of a porcine reproductive and respiratory syndrome virus genome, and the primer probe group can realize efficient detection of American strains and European strains of the porcine reproductive and respiratory syndrome virus, and is simple and convenient in detection, high in sensitivity, good in specificity, high in speed, low in false positive and capable of realizing high-throughput detection.

The invention discloses preparation and application of an antibody capable of detecting various IMP variants, and belongs to the field of biological medicine. According to the invention, a conserved epitope sequence of IMP enzyme is obtained according to comparison of amino acid sequences of all IMP variants, and an antibody of a spatial epitope is prepared through structural analysis of the epitope. After a mouse is immunized with IMP immunogen protein, hybridoma cells secreting antibodies are obtained through multi-technology platform screening, specific antibody pairs MK35G9 and MK56G2 which are high in affinity and can be combined with multiple IMP variants are obtained after sequencing, and corresponding CDR is obtained through analysis. According to the present invention, the IMP enzyme detection product development is performed based on the immunochromatography platform, such that advantages of high sensitivity, good specificity, covering of a variety of IMP variants, and rapid and convenient detection are provided, and important help is provided for clinical auxiliary diagnosis and medication guidance.

The invention discloses a nested MIRA primer combination for detecting hantavirus HTNV type and hantavirus SEOV type. The nested MIRA primer combination comprises a peripheral primer, an inner primer and a probe for detecting the hantavirus HTNV type and the hantavirus SEOV type, the peripheral primers for the hantavirus HTNV type are as shown in SEQ ID NO: 1 and SEQ ID NO: 2, the inner primers are as shown in SEQ ID NO: 3 and SEQ ID NO: 4, and the probe is as shown in SEQ ID NO: 5; the peripheral primers for the hantavirus SEOV type are as shown in SEQ ID NO: 6 and SEQ ID NO: 7, the inner primers are as shown in SEQ ID NO: 8 and SEQ ID NO: 9, and the probe is as shown in SEQ ID NO: 10; the primer disclosed by the invention can complete nested multienzyme constant-temperature nucleic acid rapid amplification within 30 minutes at the temperature of 39 DEG C, is simple and convenient to operate, has lower requirements on instruments, equipment and professional skills of operators, and can achieve the lower detection limit of 14.2 copies / mu L.

The invention discloses a detection system for simultaneously detecting circle virus poultry source type 1 and circle virus human source type 1 and application of the detection system, and relates to the technical field of biological detection. The detection system comprises an RAA amplification primer pair of GyVg1, an RAA amplification primer pair of GyH1, a 12-crRNA-3 (12-crRNA-3), a 13-crRNA-2 (13-crRNA-2), an ssDNA (single-stranded deoxyribonucleic acid) probe, an ssRNA (single-stranded ribonucleic acid) probe, an LbCas12a nuclease, an LbuCas13a nuclease and a T7 transcriptase. According to the present invention, based on GyVg1 and GyH1 conserved genes, the specific crRNA and the RAA primer are respectively designed, and the RAA amplification technology and the CRISPR-Cas fluorescence detection system are combined so as to establish the detection system for simultaneously detecting GyVg1 and GyH1; the detection system can detect two different subtypes of circle viruses at the same time, the detection efficiency is improved, and a new technical means is provided for dynamic research of virus infection.

The invention discloses an ultrasonic molecular probe. The ultrasonic molecular probe comprises a targeted contrast agent and siRNA of fibronectin, wherein the targeted contrast agent and the siRNA are connected through G0-C14; the targeted contrast agent comprises gas vesicles covalently linked with annular polypeptide, and the annular polypeptide targets an atherosclerotic plaque region. The cyclic polypeptide can be combined with the outer structural domain B of fibronectin with high specificity, and the atherosclerotic plaque extracellular matrix is rich in the outer structural domain B of the fibronectin, so that enough targeted ultrasonic contrast agent can be quickly combined in the plaque region to generate strong signal enhancement in ultrasonic imaging, and on the basis, the targeted ultrasonic contrast agent can be quickly combined with the plaque region to generate strong signal enhancement in ultrasonic imaging. The ultrasonic molecular probe releases siRNA for atherosclerotic plaque treatment, and atherosclerotic plaque diagnosis and treatment integration is achieved. The invention further discloses a preparation method of the ultrasonic molecular probe and application of the ultrasonic molecular probe in preparation of atherosclerotic plaque diagnostic agents, drugs and drug delivery agents.

The invention provides a construction method and application of an angelica sinensis potion fingerprint spectrum, and belongs to the technical field of drug detection.The construction method includes the steps that firstly, an angelica sinensis potion is used for preparing a test solution, then 0.08-0.12 wt% phosphoric acid aqueous solution (A)-acetonitrile (B) serves as a mobile phase, and the angelica sinensis potion fingerprint spectrum is obtained through high performance liquid chromatography detection, a standard fingerprint spectrum is established and is used for quality evaluation or control in the whole process of research / development / production / clinical application of the angelica sinensis potion. According to the construction method of the fingerprint spectrum of the angelica sinensis potion, main medicinal material components in the angelica sinensis potion can be detected only through high performance liquid chromatography, so that the quality condition of the angelica sinensis potion is reflected more comprehensively.

The invention provides a construction method of fingerprint spectrums of katsumadai medicinal material, standard decoction and formula granules thereof, which comprises the following steps: preparing a mixed reference solution from alpinetin, pinocembrin, cardamonin and alnudinone; preparing a test solution from the alpinia katsumadai medicinal material, the standard decoction or the formula granules; respectively detecting the reference solution and the test solution by adopting a high performance liquid chromatography, and constructing fingerprints of the alpinia katsumadai medicinal material, the standard decoction and the formula granules of the alpinia katsumadai medicinal material; wherein the conditions of the high performance liquid chromatography are as follows: a mixed solution of acetonitrile and methanol is taken as a mobile phase A, a phosphoric acid aqueous solution with the volume concentration of 0.08%-0.12% is taken as a mobile phase B, and a gradient elution procedure is adopted. The method has the advantages of being simple, stable and high in specificity, and a comprehensive, effective and rapid evaluation method is provided for quality control of alpinia katsumadai medicinal materials, standard decoction and formula granules of the alpinia katsumadai medicinal materials.

The invention discloses a method for detecting the contents of VMA, HVA and 5-HIAA in urine by using liquid chromatography-tandem mass spectrometry. The method comprises the following steps: S1, removing VMA, HVA and 5-HIAA in urine to obtain a blank urine matrix; s2, weighing VMA, HVA and 5-HIAA solid powder, fixing the volume with methanol, mixing the dissolved liquids, diluting with methanol-water to obtain a VMA, HVA and 5-HIAA working solution, and finally diluting step by step with a blank urine matrix to obtain a calibration product; s3, respectively taking a urine sample to be detected, the calibration substance and the quality control substance, adding the mixed internal standard working solution of VMA, HVA and 5-HIAA isotopes, uniformly mixing, centrifuging, and taking supernate; s4, separating the supernate by using liquid chromatography; s5, the separated supernate enters a mass spectrum to be detected, and the content of VMA, HVA and 5-HIAA in the urine sample to be detected is obtained. The detection method disclosed by the invention is simple and rapid in pretreatment, high in specificity, low in reagent consumption cost, free of environmental pollution, small in required sample size, easy in high-throughput operation and suitable for being developed into a kit for conventional clinical detection.

The invention discloses a construction method and application of a ratio-dependent electrochemical luminescence (ECL) biosensor based on an antifouling red cell membrane, and belongs to the technical fields of photoelectric analysis, biosensing and interface antifouling. In the invention, the erythrocyte membrane with super-hydrophilicity and low membrane protein content is used as an efficient antifouling interface, and the good biocompatibility of the erythrocyte membrane avoids the influence of a chemical antifouling coating on the biological activity of the ECL sensor. Meanwhile, the electropolymerized polyaniline nanowire is embedded into the antifouling erythrocyte membrane, so that the chemical stability and the conductivity of the antifouling erythrocyte membrane are enhanced, and the application of the antifouling erythrocyte membrane in ECL analysis is expanded. C3N4 is introduced as an interior label for eliminating background signals and external interference, so that the detection precision of the biosensor is improved. On the basis, the constructed ECL antifouling biosensor realizes ultra-sensitive detection of human breast cancer cells MCF-7 in human serum, and the detection limit is as low as 3 cells / mL.

The invention discloses an SERS (Surface Enhanced Raman Scattering) sensor for detecting diabetic retinopathy serum biomarkers miR-21 and miR-152 as well as a preparation method and application of the SERS sensor, and belongs to the technical field of biomedicine. The preparation method comprises the following steps: self-assembling AuNHs through an oil-water interface to obtain an AuNHs array, and modifying an aptamer chain cDNA on the surface of the array to obtain an SERS (Surface Enhanced Raman Scattering) substrate; the method comprises the following steps: modifying a Raman signal molecule 4-MBA and a nucleic acid aptamer complementary chain H1 to the surface of AuNTs to obtain an SERS probe 1; a Raman signal molecule DTNB and a nucleic acid aptamer complementary chain H2 are modified to the surface of AuNTs, and an SERS probe 2 is obtained; and combining the prepared SERS probe with a complementary basic group in the nucleic acid aptamer chain cDNA on the SERS substrate, and fixing on the AuNHs array to construct the SERS sensor.

The invention discloses a detection method and application of whole genome 5-hydroxymethylcytosine, and relates to the technical field of biochemical detection. Comprising the following steps: 1, closing a 3 '-OH end of DNA to be detected; 2, preparation of glycosylated 5hmC DNA (Deoxyribose Nucleic Acid); 3, carrying out TdT mediated DNA (Deoxyribonucleic Acid) amplification; and 4, fluorescence measurement. According to the invention, a homogeneous and non-template dependent type signal amplification method for detecting the 5-hydroxymethylcytosine in the whole genome is constructed. The method is based on a specific enzyme catalytic reaction and a multiple signal amplification strategy, and has the characteristics of high sensitivity, good specificity, no need of a separation step, no sequence dependence in amplification and capability of detecting 5hmC at any position in a whole genome. In addition, accurate and sensitive detection of actual clinical samples can be realized.

The invention relates to the technical field of microbial identification, in particular to an identification method of a streptococcus thermophilus CICC 6063 strain and development and application of a strain level primer of the streptococcus thermophilus CICC 6063 strain. The invention provides an application of a nucleotide sequence as shown in SEQ ID NO: 1 in identification of a streptococcus thermophilus CICC 6063 strain. The invention provides an identification method of a streptococcus thermophilus CICC 6063 strain. The method comprises the following steps: extracting genome DNA of a to-be-detected strain, designing a primer aiming at a nucleotide sequence shown in SEQ ID NO: 1, amplifying the genome DNA, and judging whether the to-be-detected strain is the streptococcus thermophilus CICC 6063 strain or not by comparing the length of an actual amplification product with the length of a target amplification product and the condition of polymorphic site basic groups. The invention provides a simple and rapid method for identifying a streptococcus thermophilus CICC 6063 strain by utilizing a specific conserved gene sequence of streptococcus. The primer pair provided by the invention can be used for rapidly and accurately identifying the CICC 6063 strain on the strain level.