538 results about "Nucleic acid detection" patented technology

The invention relates to the technical field of nucleic acid detection, in particular to a product and a method for detecting bifidobacterium longum subsp. Longum 6-1. Bioinformatics analysis is carried out based on a whole genome sequence of the bifidobacterium longum subspecies longum 6-1, the primer pair and the primer probe combination are obtained through optimization, and the primer pair and the primer probe combination have good specificity and sensitivity in the aspect of detecting the bifidobacterium longum subspecies longum 6-1; the bifidobacterium longum subsp. Longum 6-1 and other strains (including other same subsp. Longum strains, other same strains and other strains) can be quickly and accurately distinguished, and the strain level quantitative detection of the strain in a complex biological sample (such as excrement) can be realized.

The invention discloses a multi-subtype influenza A virus nucleic acid detection kit and a detection method, and relates to the technical field of medical detection.The kit comprises a group of reverse transcription RT-RPA amplification primer pairs used for amplifying influenza A virus M gene target sequences, the reverse transcription RT-RPA amplification primer pairs are designed for specific areas of subtypes H1N1, H2N2, H3N2, H5N1 and H7N9, and the reverse transcription RT-RPA amplification primer pairs are designed for specific areas of subtypes H1N1, H2N2, H3N2, H5N1 and H7N9; the crRNA library comprises a plurality of crRNA molecules which are respectively complementary with different conserved regions in the M gene target sequence and is used for guiding Cas12a protein to carry out specific recognition; a Cas12a protein reagent; the reporter molecule can be subjected to non-specific cleavage by the Cas12a protein; the enzyme preparation, the buffer solution and the nucleotide substrate are required by RT-RPA reaction. The invention aims to realize synchronous, rapid, high-sensitivity and high-specificity detection of various subtype influenza A viruses.

The invention relates to the technical field of biological detection, and discloses a mycoplasma pneumoniae nucleic acid rapid detection method, which comprises the following steps: mixing a sample to be detected with a multifunctional pretreatment buffer solution containing tris (hydroxymethyl) aminomethane, potassium acetate, polyethylene glycol, saponin, spermine tetrahydrochloride and tris (2-carboxyethyl) phosphine hydrochloride, and carrying out cracking treatment; a cracking mixed solution is obtained; redissolving the enzymatic isothermal amplification basic reagent, the specific primer and the probe to obtain a redissolving amplification system; sucking the supernatant of the cracking mixed solution, adding the supernatant into a redissolving amplification system, and adding a magnesium acetate solution to obtain a reaction mixed solution to be detected; and carrying out isothermal amplification and signal acquisition to obtain a detection conclusion. The multifunctional pretreatment buffer solution is used for cracking the to-be-detected sample, sample liquefaction, cracking and nucleic acid release are completed in a single system, the nucleic acid extraction and purification step of centrifugal column adsorption or magnetic bead elution is avoided, and the operation process is simplified.

The invention relates to the technical field of medical diagnosis, and discloses an integrated nucleic acid detection method and device based on a micro-fluidic chip, the integrated nucleic acid detection device comprises a micro-fluidic chip module, a liquid control module, a temperature control module, an optical detection module and a data processing module, and the multiple modules work cooperatively to realize full-process automatic detection of a sample from splitting decomposition to detection; the micro-fluidic chip module comprises a sample treatment area, a nucleic acid extraction and purification area, a nucleic acid amplification area, a nucleic acid detection area and a waste liquid collection area which are connected through a micro-channel, and a valve is arranged at a key node. Through a fluid control mode combining centrifugal force and air pressure driving and valve combination of non-complex micro-pump micro-valves such as the dissolvable membrane valve, the temperature sensitive valve and the capillary force flow limiting valve, full-flow fluid sequential control can be achieved without integrating a plurality of micro-pump micro-valves; the problems of complex structure, high manufacturing difficulty and high cost caused by integration of multiple micro-pumps and micro-valves of the conventional pressure-driven micro-fluidic chip are solved.

The invention belongs to the field of nucleic acid detection and molecular biology, and particularly relates to a method and a kit for eliminating false positive results in nucleic acid amplification reaction. According to the method, after a nucleic acid amplification reaction (taking recombinase polymerase amplification, namely RPA, for example) is completed, a target amplification product is subjected to selective enzyme digestion by utilizing restriction endonuclease, non-target products (such as primer dimers and non-specific amplification products) are not cut, and meanwhile, the non-target products which are not subjected to enzyme digestion are removed by combining solid-phase separation, so that the target amplification product is obtained. Therefore, the false positive result is eliminated. The invention also discloses a kit containing the restriction enzyme. According to the method, the specificity and the signal-to-noise ratio of a nucleic acid amplification reaction, especially RPA, are remarkably improved, the cost is low, operation is easy, the method is compatible with an existing technical platform, the method is suitable for detection scenes such as clinical molecular diagnosis, environmental monitoring and food safety, and the problem of misjudgment caused by false positive signals is effectively avoided.

The invention discloses an amplification chip and an amplification analysis device, and belongs to the field of nucleic acid detection, the amplification chip comprises a chip main body provided with a flow channel, the amplification chip also comprises a temperature control assembly, the temperature control assembly comprises at least one heating member, the at least one heating member heats a part of the flow channel, so that the flow channel forms at least two areas with different temperatures, and the heating member is used for heating the flow channel. The reagent reciprocates in two regions with different temperatures, so that the reaction time of the reagent is shortened, and the detection sensitivity of the amplification chip is improved.

This invention provides a CRISPR / Cas12a fluorescence detection method for FocTR4 based on magnetic bead-ssDNA-HRP probes, belonging to the field of nucleic acid detection technology. This invention involves isothermal amplification of the FocTR4 target sequence using RPA primers; then, a magnetic bead-ssDNA-HRP probe is prepared; and the Cas12a / crRNA complex, RPA amplification product, and probe are co-incubated. The activated Cas12a cleaves the probe, releasing HRP. After magnetic separation, a colorimetric solution containing OPD is added, and the presence and concentration of FocTR4 are determined by detecting the fluorescence signal. This invention combines the high efficiency of CRISPR / Cas12a cleavage with the high sensitivity and strong specificity of RPA technology, achieving highly sensitive and specific detection of FocTR4, with a linear detection range of 10. 2 ~10 7 With a detection limit as low as 70 CFU / mL, it can quickly and accurately detect Foc TR4 in soil samples, enabling rapid on-site detection and providing reliable technical support for the early diagnosis and control of banana wilt disease.

The invention relates to a normal temperature amplification-CRISPR Cas12a one-pot method detection system and a nucleic acid detection method based on the detection system. The detection system consists of a normal-temperature nucleic acid amplification system (unit I) which focuses on target signal amplification and a CRISPR Cas12a system (unit II) which focuses on target recognition and signal output. By means of a self-driven and deoxyribozyme-mediated reaction equilibrium transfer strategy, the equilibrium in the initial stage of the reaction tends to the unit I, and the equilibrium in the middle and later stages of the reaction tends to the unit II, so that the compatibility of the two units is remarkably improved. By means of the system, high-sensitivity and high-specificity portable one-pot detection of nucleic acid targets can be achieved.

The present invention relates to specific natural or artificial RNA or DNA / RNA substrates for cleaving by a Cas nuclease. The invention furthermore relates to a complex comprising the specific artificial RNA or DNA / RNA substrate, at least one of a Cas nuclease enzyme and at least one preselected guide RNA binding to at least one target RNA. The present invention also relates to methods for cleaving the natural or artificial RNA or DNA / RNA substrate and methods for detecting at least one target RNA in a cell, tissue, cellular nucleus, and / or sample using the substrate or for eliminating a cell expressing the at least one target RNA.

The invention provides a novel nuclease and a method for gene editing and nucleic acid detection by using the nuclease. In some embodiments, the disclosure provides uses of the programmable nuclease. The disclosure also provides compositions, systems, and methods comprising the programmable nuclease.

The present application relates to the technical field of nucleic acid detection, in particular to a RNA pathogen chemiluminescence detection method based on nucleic acid structure and sequence characteristics. The present application provides a pathogen RNA rapid detection technology based on the reporter molecule cyclization combined with rolling circle amplification according to the secondary structure and sequence characteristics of RNA, which is suitable for high-sensitivity and high-specificity detection of RNA pathogens such as respiratory viruses. The present application innovatively combines the target structure characteristics with sequence specificity, breaks through the traditional sequence-dependent isothermal amplification technology, realizes the highly specific reporter molecule cyclization, significantly improves the detection reliability, and does not require complex instruments. After the swab sample is collected, the pathogen RNA is rapidly released by using a nucleic acid releasing agent, without complex purification steps, and the reading can be realized through a common chemiluminescence instrument, which is suitable for primary site detection.

Provided herein is a nucleic acid detection system comprising a detection CRISPR system having an effector protein and one or more guide RNAs each designed to bind to corresponding target molecules that are diagnostic for a tick-borne disease state; and an RNA-based masking construct. In some embodiments, the detection system of may comprise i) two or more CRISPR systems, each CRISPR system comprising an effector protein and a guide RNA designed to bind to a corresponding target molecule that is diagnostic for a tick-borne disease state; and ii) a set of detection constructs, each detection construct comprising a cutting motif sequence that is preferentially cut by one of the activated CRISPR effector proteins. Exemplary tick-borne detectable microbes include Babesia microti, Anaplasma phagocytophilum, and Borrelia miyamotoi.

1. The name of the design product: full-automatic nucleic acid detection analyzer. 2. The use of the design product: the design product is used for automatic sample adding, extraction and detection analysis of nucleic acid in clinical biological samples, and is suitable for large hospital outpatient and emergency departments, medical center laboratory sites. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective view 1.

The invention belongs to the technical field of biology, and relates to a bronchoalveolar lavage fluid (BALF) preserving fluid, a kit and a cytological test method. The preservation solution is composed of TCEP, N-acetylcysteine, EDTA.2Na, trehalose, Proclin300, methanol, sodium chloride and HEPES, and all the components have a synergistic effect to achieve sample viscosity removal, oxidation resistance, corrosion prevention, cell protection and pH stabilization. The preserving fluid is suitable for morphological observation and nucleic acid detection of BALF cells, and can stably preserve samples at room temperature and maintain completeness of cell membranes and clear morphological structure. The kit comprises two preparations which are mixed to form a working solution, and the working solution can be directly used for sample preservation on a sampling site. The cytological test method comprises the steps of sample collection, preservation, centrifugal slide preparation, Wright-Giemsa staining and microscopic observation. Experimental results show that preservation solutions of different formulas are compatible with a chromosome system, the cell morphology is kept stable within 3 days, the dyeing effect is good, and it is proved that the preservation system can remarkably improve the preservation stability and detection reliability of BALF samples and is suitable for cytology and molecular diagnosis application of lower respiratory diseases.

The embodiment of the utility model relates to the technical field of in-vitro medical diagnosis devices, and discloses a nucleic acid detection card box and a nucleic acid detection system.The nucleic acid detection card box is provided with a reagent inlet, a reagent bag is arranged at the reagent inlet, a reagent is contained in the reagent bag, and the card box is provided with a puncture structure at the reagent inlet in a protruding mode; the reagent bag is extruded and punctured by the puncturing structure when being pressed, so that a reagent in the reagent bag enters the card box through the reagent inlet. Through the mode, the corresponding reagent bag can be arranged at the corresponding reagent inlet in advance in the preparation process of nucleic acid detection, so that the operation of repeatedly replacing the reagent bag for injection in the detection process is omitted, and the reagent bag can be easily punctured through the puncturing structure when a reagent needs to be injected; due to the arrangement of the position of the puncturing structure, the puncturing position of the reagent bag can be controlled.

The present application provides a digital PCR detection apparatus, a digital PCR quantitative detection method, a multi-volume digital PCR quantitative analysis method, a digital PCR detection method, a nucleic acid detection microsphere, a preparation method of the nucleic acid detection microsphere, a kit, and a high-throughput nucleic acid detection method.

The invention relates to the technical field of nucleic acid detection, and discloses a method and device for detecting and distinguishing radix trichosanthis, poria cocos and radix puerariae and terminal device.The method comprises the steps that genome DNA of a to-be-detected sample is extracted, a specific DNA fragment is amplified through a universal amplification primer and a specific amplification primer, and an amplification product is obtained; the method comprises the following steps: respectively constructing independent detection modules aiming at radix trichosanthis, poria cocos and radix puerariae, wherein each independent detection module comprises Cas12a protein, a test probe and specific crRNA; the specific crRNA is obtained by screening according to a PAM sequence and a detection contrast ratio. According to the invention, genome DNA of a to-be-detected sample is extracted, a universal amplification primer and a specific amplification primer are utilized, then an amplification product is added into an independent detection module, and specific crRNA is utilized to carry out specific recognition on the amplification product, so that a molecular detection system aiming at radix trichosanthis, poria cocos and radix puerariae is established, and effective distinguishing of different traditional Chinese medicinal material components is realized.

Systems and methods to detect a polynucleotide strand in a sample during the early stage of amplification cycles. A metal-insulator-metal (MIM) electrode unit with two metal electrodes separated by a dielectric layer may be utilized as a nanogap sensor. Primers for amplifying a target polynucleotide strand may be immobilized on the surface of the dielectric layer. Polymerases, nucleotides, templates and primers may be added to the sample. At least one type of nucleotide may be labeled with a redox label that may enhance electron transport through the nanogap via tunneling and / or diffusion-based redox cycles. Signals (e.g., voltage pulses) may be directed through each of the electrodes. Current values from each electrode may be measured to establish a baseline signal. Alterations in the baseline signal may indicate the presence of target polynucleotide strands. The time to detect an alteration in the baseline signal may facilitate quantification of target polynucleotide strands.

The invention discloses a detection kit based on orthogonal dual-channel label-free CRISPR-Cas and application of the detection kit, relates to the technical field of nucleic acid detection, and constructs an orthogonal dual-channel response system by utilizing the differentiated cleavage activity of Cas12a targeted DNA and Cas13 targeted RNA to a substrate. A Cas12a pathway is reported by utilizing protoporphyrin IX to be compounded with G-quadruplex G4DNA, and a Cas13a pathway is reported by utilizing DFHBI to be compounded with a Broccoli RNA structure, so that label-free signal transduction is realized. Through accurate screening of fluorophores, channel specificity is ensured, and optical signal crosstalk is eliminated. Mycobacterium tuberculosis (MTB) and respiratory syncytial virus (RSV) are used as model pathogens for verification, and a multiple recombinase polymerase amplification (RPA) technology is combined, so that the detection sensitivity on a synthetic target reaches a single molecule level. The system has high specificity, and even if the concentration of a non-target pathogen is increased by 100 times, no cross reaction exists.

The invention belongs to the technical field of microbial nucleic acid detection and bioluminescence analysis, and discloses a bioluminescence nucleic acid detection method and kit based on DNA-RNA hybrid specific recognition. The kit comprises a streptavidin coated solid phase carrier, a biotinylated DNA capture probe, a signal enhancement DNA, a luminescence report fusion protein and a luminescence substrate thereof, the luminescent reporter fusion protein comprises a DNA-RNA hybrid binding domain and a luminescent enzyme domain. According to the method, target RNA is captured through a solid phase, a DNA-RNA hybrid is formed, the luminous report fusion protein is specifically combined with the DNA-RNA hybrid, a bioluminescence signal is generated after a luminous substrate is added, and the target RNA is detected. In the embodiment, an amplification step is not adopted, so that the method has the advantages of simplicity and convenience in operation, rapidness in detection, lower background and better specificity, and can be used for rapid detection of microorganisms.

A removable cartridge to be used in a system for extracting and detecting nucleic acids from heterogeneous samples includes a plurality of reservoirs defining at least a first wash buffer reservoir for holding a first wash buffer and a microfluidic assembly configured to attach to the plurality of reservoirs. The microfluidic assembly includes at least one sample reservoir and a nucleic acid extraction matrix in fluid communication to an automated sample preparation (ASP) reservoir through a first flow channel defined by the microfluidic assembly. An assay chamber is in fluid communication with a third flow channel and with the waste reservoir through a fourth flow channel such that a labeled nucleic acid-containing sample flows through the assay chamber and then to the waste reservoir, wherein vibration-driven mixing agitates fluids while present in the assay chamber. Finally, a nucleic acid-detecting microarray module is positioned in the assay chamber.

The invention relates to the technical field of nucleic acid detection, in particular to an RNA pathogen chemiluminescence detection method based on a nucleic acid structure and sequence characteristics. The invention provides a pathogen RNA rapid detection technology which is based on RNA secondary structure and sequence characteristic reporter molecule cyclization and is combined with rolling circle amplification, and is suitable for high-sensitivity and high-specificity detection of RNA pathogens such as respiratory viruses. Target structure characteristics and sequence specificity are creatively combined, a traditional sequence-dependent isothermal amplification technology is broken through, high-specificity reporter molecule cyclization is achieved, the detection reliability is remarkably improved, complex instruments are not needed, after swab samples are collected, pathogen RNA is rapidly released through a nucleic acid releasing agent, complex purification steps are not needed, and the method is suitable for large-scale popularization and application. And reading can be realized through a common chemiluminiscence instrument, and the method is suitable for base-level field detection.

The invention relates to the technical field of nucleic acid detection, and particularly provides a CRISPR-Cas12a cascade signal amplification system and application thereof. The system comprises a first-stage sensing unit, a second-stage sensing unit and a third-stage sensing unit, wherein the first-stage sensing unit is configured to recognize a target to be detected and activate the trans-cleavage activity of first-stage Cas12a protein; an engineered scaffold RNA composed of an RNA sequence and a DNA sequence is cut, the obtained RNA sequence serves as an activator of a second-stage Cas12a protein, and the second-stage Cas12a protein obtains trans-cleavage activity, so that a fluorescence report probe is cut, and a signal is output. According to the system, an activation event of a first Cas12a is converted into an exponential signal output unit of a second Cas12a, and a cascade amplification network is formed. According to the design, background signal leakage is eradicated from a physical mechanism, and the detection sensitivity is remarkably improved. The detection method provided by the invention is low in cost, high in sensitivity and high in reaction speed.

The invention discloses a real-time detection system and a monitoring and early warning method for African swine fever virus. The system comprises a latex microsphere labeled immunochromatography antibody detection card, a portable MINI-PCR instrument and a freeze-dried microsphere type fluorescent PCR detection reagent, the detection card takes the recombinant ASFV p30 protein as a target spot, antibody detection is completed within 10 minutes, the lowest detection limit is 1: 1024, and the specificity is greater than 99%; the PCR reagent adopts a double-primer double-probe design, and the lowest detection limit is 1 copy / microliter. According to the invention, through a collaborative process of antibody screening, nucleic acid detection and result evaluation, ASFV total infection cycle detection is realized, epidemic situation discovery time is advanced to 1-3 days after infection, and early warning response time is shortened by more than 60%; the system is simple to operate and low in cost, does not need professionals and laboratories, can be deployed in basic-level scenes such as pig farms and slaughter houses, effectively solves pain points of an existing detection system, and provides key technical support for prevention and control of African swine fever.

The application relates to the technical field of molecular biology detection instruments, and discloses a rapid nucleic acid analysis system which comprises a frame mechanism, a sample pretreatment mechanism and a PCR detection mechanism. A pipetting module and a nucleic acid extraction module are co-assembled on the same support plate and independently move, cooperate with a reagent carrier module moving along the Y direction, and realize integrated operation of sample pipetting, nucleic acid extraction and transfer; a PCR tube support plate is floatingly arranged through an elastic support structure, and under the action of a compression module, the PCR tube is tightly combined with a heating hole of a temperature control module; an optical detection module adopts a turntable type light filtering switching and light blocking structure, and reduces channel light interference. The system has high integration degree, small volume, is favorable for improving nucleic acid detection efficiency, temperature control uniformity and detection accuracy.

Provided are a formulation for enhancing nucleic acid release efficiency in a sample and a use thereof. Specifically, the formulation comprises a preservation solution, and further comprises grinding filter beads. Using the formulation of the present application to treat a sample can enhance the nucleic acid release efficiency of the sample. With the assistance of ultrasound and / or heating, the nucleic acid release efficiency of the sample is further improved, thereby simplifying sample collection and processing procedures.

The embodiment of the invention discloses a micro-fluidic chip and application thereof in nucleic acid detection.The micro-fluidic chip comprises a chip part, the chip part comprises a substrate, and the substrate comprises a top layer, a middle layer and a bottom layer; the substrate is provided with a microfluidic channel; three-dimensional patterns are arranged on the upper surface and the lower surface of the middle layer, and the pattern on the upper surface of the middle layer and the top layer define an upper-layer microfluidic channel in the microfluidic channel; the pattern on the lower surface of the middle layer and the bottom layer define a lower-layer microfluidic channel in the microfluidic channel; the extending directions of the upper-layer microfluidic channel and the lower-layer microfluidic channel are parallel to the plane of the substrate; the middle layer comprises a through hole used for forming a vertical microfluidic channel, the extension direction of the vertical microfluidic channel is perpendicular to the plane where the substrate is located, and at least part of the vertical microfluidic channel is used for communicating the upper-layer microfluidic channel with the lower-layer microfluidic channel. According to the embodiment of the invention, an overpass type microfluidic channel can be formed, the channel density can be improved, and a flexible communication relation can be constructed to meet complex reaction requirements.

The invention belongs to the field of molecular detection, and particularly relates to a nucleic acid detection analyzer and a nucleic acid detection method.The nucleic acid detection analyzer uses special detection consumables, and the detection consumables adopt closed design and can prevent pollution. In the nucleic acid extraction and storage process, after the detection consumables are placed in the carrying seat, transfer and liquid adding are not needed, the sample liquid in the sample tube is subjected to ultrasonic splitting decomposition through the ultrasonic module, nucleic acid is extracted through a one-step method, and then the nucleic acid solution and the reaction liquid in the reaction tube are mixed through the gland module; nucleic acid in the reaction tube is subjected to heating amplification and optical detection by the PCR detection module, and finally, a detection result is processed, analyzed and output, so that manual operation is reduced, and sample feeding and result outputting are realized. The nucleic acid detection method based on the nucleic acid detection analyzer is simple to operate, sample pretreatment, nucleic acid extraction and detection processes are integrated, the professional requirement on operators is reduced, and the application scene of the nucleic acid detection analyzer is greatly expanded.

The application relates to a sealed integrated device for magnetic bead method nucleic acid detection and application thereof, the integrated device comprising a card box, a pipettor and a telescopic elastic sealing cover, the card box comprising an upper cover, a sealing gasket and integrated reagent storage tubes, the integrated reagent storage tubes being sequentially fixedly connected on a connecting body by opening parts of several test tubes, detection reagents being previously filled into different test tubes of the integrated reagent storage tubes, and then the sealing gasket and the upper cover being covered to realize reagent sealing in the card box; the lower part of the telescopic elastic sealing cover being detachably and sealingly connected with the upper part of the card box, and the pipettor being arranged in the telescopic elastic sealing cover. The sealed integrated device can be filled with reagents in advance, automatically operated in a mechanical mode, realizes POCT detection, has a wide application range, is convenient to upgrade to micro-fluidic chip detection, has a simple structure, is easy to match with existing detection facilities, and can be rapidly popularized and applied.

The invention relates to the technical field of biomedical detection, and discloses a treponema pallidum constant-temperature nucleic acid detection sample pretreatment device which comprises a treatment box and an integrated support carrier serving as the whole device, and a placement groove for positioning a detection test tube and a storage tube for storing a standby magnetic bar are arranged in the treatment box. Regular layout and protection of parts are realized; the synergistic cracking assembly is integrally mounted on the left side in the treatment box and is used for cracking the tough outer membrane of the treponema pallidum through the synergistic effect of mechanical oscillation and ultrasound so as to ensure full release of nucleic acid; the constant-temperature treatment assembly is in linkage installation with the collaborative cracking assembly and is used for providing a precise constant-temperature environment for sample cracking and a subsequent nucleic acid binding process. A synergistic system of treponema pallidum nucleic acid pretreatment is constructed by additionally arranging the synergistic lysis assembly and matching with the constant-temperature treatment assembly, and the synergistic system and the constant-temperature treatment assembly form a multi-dimensional lysis acting force, so that the technical problems that the treatment efficiency of stubborn pathogens is low and nucleic acid release is incomplete in a traditional single lysis mode are effectively solved.