23 results about "Enzyme free" patented technology

The utility model discloses automatic enzyme-linked immunoassay detection equipment which comprises a processing module and a washing liquid module, and the processing module comprises a pipetting module, a clamping jaw module and an operation processing area, the operation treatment area is provided with a reaction container, a cleaning position, a cleaning head matched with the cleaning position, an oscillation incubation area, a sample adding area, a consumable loading area, a reagent loading area and an enzyme immunoassay detection area, the cleaning head comprises a liquid injection needle and a liquid suction needle, and the pipetting module and the clamping jaw module can be driven to move along the operation treatment area. The pipetting module can transfer a sample liquid or a reagent to the reaction container, the clamping jaw module can move to the reaction container to perform cover opening and closing operation and can transfer the reaction container into a cleaning position, and the washing liquid module can be in fluid communication with the liquid injection needle and the liquid suction needle to complete cleaning of the reaction container. The detection equipment provided by the utility model can realize a series of full-automatic enzyme immunoassay detection such as sample transfer, reagent transfer, enzyme immunoassay reaction, cleaning and enzyme immunoassay detection.

The present invention is applicable to the field of electrochemical sensors and provides a method for preparing an enzyme-free electrochemical sensor with a modified electrode and its application, comprising the following steps: drawing a capillary quartz tube into a needle-shaped nanoquartz tube, uniformly distributing carbon nanoparticles on the tip of the nanoneedle tip to form a carbon nanotube (CNE); dissolving HAP, sodium hexametaphosphate, and potassium chloride in deionized water at a concentration ratio of 0.01 to 0.1:1:1, and ultrasonically forming a dispersion; placing the CNE as a working electrode, Ag / AgCl as a reference electrode, and a platinum wire as an auxiliary electrode in an electrolytic cell containing a HAP suspension solution, applying an external positive voltage to the working electrode CNE, and completing constant potential deposition to prepare an nHAP-CNE modified sensor. The sensor prepared based on the nHAP modified electrode has the advantages of high sensitivity, low sample requirement, high biological activity, and suitability for in vivo cell detection in detecting ascorbic acid.

The invention belongs to the technical field of nucleic acid detection and mass spectrometry, and particularly relates to a nucleic acid multiple analysis and detection platform based on full DNA mass coding and normal pressure mass spectrometry detection, and the platform constructs a non-enzyme cascade amplification system completely composed of DNA molecules. A group of DNA hairpin structure chains capable of responding to a target miRNA to be detected to trigger a hybridization chain reaction are designed, a DNA enzyme core with catalytic activity is formed in a product obtained by the hybridization chain reaction of the DNA hairpin structure chains and the target miRNA to be detected, the DNA enzyme can break a substrate chain and release a DNA fragment with characteristic quality coding, and a signal is directly read by virtue of a normal pressure mass spectrum, so that the target miRNA to be detected can be detected. Parallel recognition and quantitative detection of multiple miRNAs are realized; the reaction can be spontaneously completed in a homogeneous aqueous solution system, parallel recognition and accurate quantification of multiple miRNA molecules are realized, and a novel detection method with high compatibility and high sensitivity is provided for high-throughput nucleic acid detection and multi-marker diagnosis.

The invention relates to a Na-doped nickel-based phosphate / graphene composite material, an enzyme-free glucose electrochemical sensor, a preparation method and application of the Na-doped nickel-based phosphate / graphene composite material and the enzyme-free glucose electrochemical sensor in glucose detection, and solves the technical problems that an existing enzyme-free glucose sensor prepared on the basis of Ni salt is low in detection sensitivity and slow in response speed. The preparation method comprises the following steps: firstly, sequentially adding nickel salt, sodium salt and phosphate into a graphene dispersion liquid to form a mixed solution, then carrying out hydrothermal reaction on the mixed solution, and then carrying out calcination post-treatment. The method can meet the requirements on rapid and accurate detection of glucose in the fields of clinical diabetes diagnosis, food detection and the like, and has a wide application prospect.

The present application relates to the field of bioengineering and microbial technology, and relates to a yeast surface display construction method of a double-enzyme system. AGA1 The gene promoter is replaced by a strong constitutive promoter TEF1 and a glucose-responsive promoter HXT7 ; a double-enzyme free expression vector containing a galactose-inducible promoter GAL1 driven detoxification enzyme gene is constructed, and is transformed into a modified chassis cell to obtain a double-enzyme co-display engineering bacterium; the chassis protein AGA1 is expressed in a culture medium, and YtdepA and YtdepB are induced in a galactose-induced culture medium, so that the YtdepA and YtdepB are co-displayed on the yeast surface through the coupling mechanism of AGA1-AGA2. The strong constitutive promoter TEF1 and the glucose-repressed promoter HXT7 are coupled with the galactose-inducible promoter GAL1 respectively, so that the chassis protein and the detoxification enzyme are matched in the expression timing and intensity, and the detoxification enzyme is displayed on the yeast surface.

The present application relates to aldose reductase activity analysis technical field, specifically relates to a kind of aldose reductase activity and its inhibitory effect quantitative analysis method.Local substrate concentration regulation is constructed by dividing multiple detection segments, combining bypass sampling channel, electrochemical detection cavity and compensation injection point, and the substrate environment of each detection segment is maintained consistent;Enzyme-free reference channel parallel to microfluidic reaction channel is set, and the total absorbance change amount background is separated, to obtain the real enzymatic absorbance change amount;Based on sliding time window, extract local dynamic enzyme reaction rate, combined with wall adsorption correction to eliminate the influence of enzyme activity attenuation along the way, identify stable reaction interval through local fluctuation coefficient of variation, determine local half response time and local onset time;Local onset time is used as the integration starting point, to calculate comprehensive inhibition intensity, realize inhibitor onset order and inhibition strength spatial quantitative analysis, improve the accuracy, stability and spatial comparability of detection result.

The invention belongs to the technical field of nucleic acid molecule detection, and particularly relates to an enzyme-free isothermal strand displacement cascade amplification reagent for acute respiratory pathogen detection and application of the enzyme-free isothermal strand displacement cascade amplification reagent. The detection method based on the amplification reagent is based on a nucleic acid reaction system composed of an initiation double strand and two molecular beacons, cyclic amplification is performed through a toehold-mediated strand displacement reaction trigger signal under a constant temperature condition, and end point interpretation is performed through a fluorescence signal. The method does not need enzyme and thermal cycle, is short in detection time, adapts to high-throughput operation of a porous plate, and realizes specific recognition and signal cascade amplification of target nucleic acid under a constant temperature condition, so that rapid, accurate and high-throughput detection of acute respiratory pathogens is realized on the premise of not depending on enzymatic reaction and thermal cycle equipment. In addition, the kit has the advantages of low cost, good stability and wide application range, and is suitable for rapid detection of various acute respiratory pathogens.

Described herein is a one-pot, four-enzyme cascade of enzymes, three bound to quantum dots with one enzyme free in solution, for the conversion in vitro of fumarate to 1,3-diaminopropane. The cascade operates via two distinctly different enzymatic channeling mechanisms which simultaneously function to increase the overall rate. The first three enzymes of the pathway (AspB->LysC->Asd) were able to engage in channeling in a nanoparticle displayed format, but addition of the last two enzymes to this pathway in this format (AspB->LysC->Asd->Dat->Ddc) did not result in complete channeling through the entire pathway to the final diaminopropane product. Surprisingly, replacement of the last two enzymes (Dat->Ddc) with a naturally occurring fused Dat-Ddc hybrid (Daba) provided for full channeling in this system (AspB->LysC->Asd->Daba).

The application relates to the technical field of electrochemical sensors, in particular to an enzyme-free ascorbic acid electrochemical sensing electrode based on Ni-HITP and application thereof. The electrochemical sensing electrode is a Ni3(HITP)2 / CC electrode modified by two-dimensional conjugated MOF Ni-HITP and directly used as an electrode material and a sensing interface, and is prepared by adopting a simple hydrothermal method with carbon cloth as a substrate. The electrochemical sensing electrode is used for high-sensitivity enzyme-free ascorbic acid electrochemical sensing. In a 0.1M NaOH solution, ascorbic acid sensing test is carried out by using an optimal potential of 0.55V. The electrochemical sensing electrode has excellent performances such as a large detection range, a low detection limit, high sensitivity, rapid response, high selectivity and high stability, and has great application potential.

The present invention discloses a terahertz-based, enzyme-free detection method and kit for peripheral blood free microRNAs. By designing a rational size and utilizing nanoparticles and magnetic nanoparticles to assemble into a 3D nanomagnetic network, the mismatch between the terahertz wavelength and the physical diffraction scale of the miRNAs to be detected is resolved. The surface plasmon resonance caused by the high refractive index of the nanoparticles and the electromagnetic response mechanism of the magnetic nanoparticles achieve a dual amplification effect of the THz metamaterial's resonance peak frequency shift signal, thus resolving the sensitivity issues of enzyme-free, amplification-free detection of miRNAs in biological samples. The biosensor of the present invention has high sensitivity, strong specificity, and can achieve high-throughput detection, providing a technical platform with greater clinical application prospects for highly reproducible and high-throughput detection of miRNAs.

The application belongs to the field of glucose detection, and particularly relates to an enzyme-free glucose sensor based on a hydrophobic substrate and a preparation method and application thereof. The application can realize glucose detection in various pH solution environments, reduces the dependence on high OH ‑ concentration of a solution; and the oxygen reduction reaction can be carried out at a lower reduction potential, which provides convenience for low-energy consumption use of the sensor.

The invention relates to the technical field of non-enzymatic cell dissociation, in particular to a non-enzymatic adipose tissue dissociation method and device based on hydrodynamic shearing, and the method comprises the following steps: step 1, obtaining dissociation indexes, and obtaining an optimal process parameter combination I meeting the dissociation indexes through a model based on the dissociation indexes; step 2, establishing a reference process based on the optimal process parameter combination I; step 3, judging whether a decision is input or not, if so, entering step 4, and otherwise, performing a dissociation process based on a reference process; and step 4, optimizing the reference process based on decision input to obtain an optimal process parameter combination 2, and performing a dissociation process based on the optimal process parameter combination 2.

The invention belongs to the technical field of glucose sensing, and discloses an enzyme-free glucose sensing electrode based on a porous biological membrane, a preparation method and application, and the sensing electrode comprises a graphite substrate, a conductive silver layer, an egg shell membrane layer, a platinum conductive layer and a ferroferric oxide magnetic nanoparticle layer from bottom to top. The graphite substrate and the egg shell membrane layer are conductively glued through the conductive silver layer, and the structural advantages of the three-dimensional porous microstructure of the egg shell membrane are fully reserved. The platinum conducting layer improves the conductivity of the electrode substrate, and achieves efficient electron transmission performance through the synergistic effect of the platinum conducting layer and ferroferric oxide serving as nano-enzyme. The three-dimensional porous microstructure of the egg shell membrane increases the specific surface area of the surface of the sensor electrode, can provide a stable and solidified substrate for a nano-enzyme catalytic material, greatly enriches the active sites of the sensor working electrode, and ensures rapid and accurate enzyme-free glucose sensing detection. Meanwhile, the sensing electrode has the advantages of being easy to degrade, high in biocompatibility, environmentally friendly and the like.

The present invention relates to methods for the synthesis of oligonucleotides and related compounds supported on a solid support. The inventive method provides synthetic schemes that permit the coupling of two or more oligonucleotide fragments attached to the same support, in a fashion that allows access to previously unobtainable structures, and provides a more convenient and higher-yielding approach to known oligonucleotides, including those of commercial significance.

The present invention belongs to the field of analysis and detection, and specifically relates to a biosensor for rapid detection of microcystin-LR. The present invention provides an enzyme-free biosensor for MC-LR detection based on the principles of aptamer recognition and self-assembly of hairpin probes on magnetic beads. The amplification strategy of toehold-mediated chain displacement and cyclic hybridization reaction is connected in series, and high-sensitivity detection of MC-LR is achieved by relying on a fluorescence-sensitive platform. This method simplifies the operation and reduces the cost. The entire detection process responds quickly, and the operation process can be mastered without professional training, which is convenient for rapid promotion and use. The method reaches a detection limit of up to 53fM and has good specificity. It can only specifically detect microcystin-LR, but does not specifically recognize other substances such as microcystin-LA, microcystin-YR, and microcystin-RR.

The utility model discloses an incubation module and full -automatic enzyme free instrument, wherein the incubation module includes the incubator cover, the incubation module is equipped with the placement area for placing the incubator cover, the incubator cover includes first cover, second cover, counterweight spare and heating assembly, second cover with first cover is connected to and is enclosed and forms the accommodation cavity, the counterweight spare is located in the accommodation cavity, the heating assembly includes heating spare and conducting structure, heating spare is located in the accommodation cavity, conducting structure with heating spare is connected, and at least part of conducting structure stretches out the accommodation cavity outside, when the incubator cover is located in the placement area, conducting structure can with external circuit guide to with heating spare heating. It is designed to improve the incubation efficiency while reducing the production of condensate during the incubation process, also avoid the risk of incubator cover being corroded.

The invention discloses an enzyme-free electrochemical sensor based on functionalized MOFs as well as preparation and application of the enzyme-free electrochemical sensor, and relates to the technical field of food safety detection. The sensor is constructed by compositely modifying the surface of a glassy carbon electrode with ferroferric oxide nanoparticles and an amino-functionalized metal organic framework material. By utilizing the porous structure and high specific surface area of MIL-101-NH2 and the specific coordination effect of amino groups on the surface of MIL-101-NH2 and dimethoate molecules and combining the high electron mobility of Fe3O4, the high-sensitivity and high-selectivity detection of the organophosphorus pesticide dimethoate in fruits and vegetables is realized. The sensor is enzyme-free, avoids instability of biological enzyme, has the advantages of simplicity and convenience in preparation, low cost, rapidness in detection, good stability, high reproducibility and the like, is wide in linear range and low in detection limit, is suitable for rapid and accurate detection of trace dimethoate residues in fruits and vegetables, and provides a reliable technical means for on-site screening of quality safety of agricultural products.

This invention relates to the field of aldose reductase activity analysis technology, specifically to a quantitative analysis method for aldose reductase activity and its inhibitory effect. By dividing the detection into multiple segments and combining a bypass sampling channel, an electrochemical detection chamber, and a compensation injection point to construct a local substrate concentration control system, the substrate environment of each detection segment is maintained consistently. An enzyme-free reference channel parallel to the microfluidic reaction channel is set up to separate the background of the total absorbance change, obtaining the true enzymatic absorbance change. Based on a sliding time window, the local dynamic enzyme reaction rate is extracted, and the influence of enzyme activity decay along the process is eliminated by wall adsorption correction. The stable reaction interval is identified by the local fluctuation coefficient of variation, determining the local half-response time and local onset time. Using the local onset time as the integration starting point, the comprehensive inhibition intensity is calculated, achieving spatial quantitative analysis of the inhibitor's onset order and inhibitory strength, improving the accuracy, stability, and spatial comparability of the detection results.

This invention relates to an enzyme-free biosensor based on the composite nanomaterial AuCuNPs-MWCNTs and its applications. The sensor includes a working electrode, a reference electrode, and a counter electrode. The working electrode is prepared using the following process: S1: AuCuNPs and multi-walled carbon nanotubes are dispersed in tetrahydrofuran under ultrasonication to obtain a homogeneous solution of AuCuNPs-MWCNTs; S2: Pretreatment of the Au electrode; S3: The homogeneous solution is dropped onto the Au electrode and dried to obtain an AuCu@MWCNTs / Au electrode. This enzyme-free biosensor utilizes both the electrocatalytic properties of AuCuNPs and the rapid electron transfer properties of MWCNTs to achieve ultra-high sensitivity detection of glucose and hydrogen peroxide. According to this invention, an enzyme-free biosensor with ultra-high sensitivity, good selectivity, excellent stability, and reproducibility has been constructed, showing great application potential in clinical testing.

The invention relates to the technical field of nucleic acid detection, and discloses an enzyme-free nucleic acid detection method based on single-molecule tethering, which comprises the following steps: synthesizing a single-molecule tethering probe for specifically recognizing target RNA (Ribonucleic Acid), and modifying the single-molecule tethering probe in a detection chip through combination of biotin and streptavidin; a recognition probe capable of being specifically combined with target RNA is modified on PS carboxyl microspheres through covalent combination of amino and carboxyl, and functional microspheres are prepared; adding the functionalized microspheres into a to-be-detected sample, performing room-temperature rotary incubation, introducing into a detection chip, then acquiring an image, and reading a nucleic acid detection result. According to the non-enzymatic nucleic acid detection method based on the single-molecule tethering, the target gene is enriched through the functionalized PS microspheres and introduced into the detection chip of the modified single-molecule tethering probe, after the target gene is combined with the probe, a biological signal is converted into a visual microsphere displacement signal, and then the target gene is detected through the visual microsphere displacement signal. The visual detection of the single nucleic acid molecule is realized.

The invention discloses an enzyme-free sensitive material for electrochemically and continuously monitoring lactic acid in sweat in real time as well as a preparation method and application of the enzyme-free sensitive material, and particularly relates to the field of electrochemical sensing. Comprising the following steps: mixing a CuCl2 solution and NaBH4 to obtain Cu hydrogel; the Cu hydrogel is subjected to freeze drying, and Cu aerogel is obtained; and calcining the Cu aerogel to obtain a non-enzyme sensitive material CuO aerogel, using the non-enzyme sensitive material CuO aerogel to prepare a non-enzyme sensitive chip, and monitoring the content of lactic acid in sweat. The CuO aerogel is used as a sensitive material, the CuO aerogel has a rich hierarchical porous structure and a large specific surface area, mass transfer diffusion in the electrochemical process is accelerated, rich active sites are provided, the sensitivity of sensing detection is further improved, and the lower detection limit of the CuO aerogel is reduced; in addition, the CuO aerogel has excellent stability compared with a biological enzyme in traditional lactic acid sensing, so that the problem of stability reduction caused by enzyme inactivation is avoided.

The present invention relates to a method for enzyme-free miRNA multi-target detection based on PhC-coupled FRET, belonging to the technical fields of analytical chemistry and biomedicine. The present invention designs photonic crystals PhC with different reflected light colors as carriers for multi-target detection, combined with hairpin sequences that specifically recognize targets, and can synchronously optically detect multiple miRNAs, such as the detection of overexpressed miRNA-21, miRNA-155, and miRNA-10b in breast cancer. Specifically, by immobilizing the fluorescent donor hairpin chain on the PhC, when the target miRNA is specifically recognized, the miRNA triggers an enzyme-free amplification reaction, resulting in the formation of a double strand between the fluorescent donor hairpin and the fluorescent acceptor hairpin, thereby generating fluorescence resonance energy transfer FRET. By correlating the FRET ratio with the miRNA concentration, the detection of the miRNA concentration in the sample can be achieved, and the specific miRNA is determined by the reflected light colors of different PhCs. This method has high sensitivity, sequence specificity, and multi-target detection ability, and can be used for the determination of miRNA content in untreated serum.

The invention discloses Fe / nitrogen co-doped carbon microspheres as well as a preparation method and application thereof in uric acid detection, and belongs to the technical field of analytical chemistry. The preparation method comprises the following steps: dissolving a carbon source in water, stirring, carrying out hydrothermal reaction, and washing and drying an obtained solid product to obtain carbon microspheres; the preparation method comprises the following steps: putting the carbon microspheres, ferric salt and a nitrogen source into water, stirring and drying, heating, cooling, washing and drying to obtain the Fe / nitrogen co-doped carbon microspheres. The linear response range of the prepared carbon microspheres to uric acid reaches 14-454 M. In an actual urine test, the adding standard recovery rate is stabilized at 95.5-102.2%, the relative standard deviation is lower than 2.5%, high-sensitivity and high-selectivity detection of uric acid can be realized, and a new thought is provided for developing a low-cost and high-performance enzyme-free biosensor. The uric acid detection method does not need biological enzyme participation, has the advantages of low cost, good stability, simplicity and convenience in operation and the like, and provides a new technical scheme for clinical uric acid detection.