50 results about "Thermal cycler" patented technology

The invention discloses a pathogenic bacteria detection kit and a detection method based on RPA-CRISPR / Cas12a. According to the detection method, multiple pathogenic bacteria are synchronously amplified by adopting an RPA technology, and then amplification products are detected by adopting a CRISPR / Cas12a detection system, so that synchronous and rapid detection of the multiple pathogenic bacteria is realized; the pathogenic bacteria detection kit based on RPA-CRISPR / Cas12a uses an RPA technology to perform isothermal amplification on a target gene, does not need to depend on a large-scale thermal cycle instrument, has low experimental requirements, can realize synchronous amplification of multiple pathogenic bacteria molecules within 10-20 min, and can realize synchronous real-time detection of multiple pathogenic bacteria by combining a CRISPR / Cas12a system, so that the detection kit has the advantages of high sensitivity, high specificity, high sensitivity, high sensitivity and the like. The whole pathogen detection process consumes less time and does not exceed 1h; the detection sensitivity is also relatively high and reaches 100 CFU / mL; and synchronous and rapid detection of various pathogenic bacteria in hospital infection is realized.

The invention relates to the technical field of biological detection, in particular to an LAMP (loop-mediated isothermal amplification) detection kit for clostridium difficile tcdA and tcdB. The kit comprises a specific LAMP primer group, an LAMP amplification reaction mixed solution, a lysis solution, a positive reference substance, a negative reference substance and an operation instruction. The system is based on a loop-mediated isothermal amplification (LAMP) principle, nucleic acid detection of toxin-producing genes tcdA and tcdB of clostridium difficile is realized under the condition of 61-65 DEG C, and a detection result is judged through colorimetric change of HNB or a fluorescence signal; according to the system, an amplification reaction can be completed without a thermal cycler, the reaction time is about 50 minutes, and rapid detection can be realized on site; the method disclosed by the invention has the advantages of high sensitivity, strong specificity, simplicity and convenience in operation and low equipment dependence, and is suitable for detecting the toxin-producing gene of the clostridium difficile in medical institutions, primary laboratories and public health monitoring.

The invention relates to the field of agricultural pest drug resistance detection, and provides a detection primer and method for chilo suppressalis ryanodine receptor I4758M mutation and application. According to the detection primer, a mutation recognition site is designed at the 3'tail end of an inner primer BIP, specific detection is achieved through the key base difference (A / G) of a wild type and a mutant type, and the primer group comprises an outer primer pair F3 / B3, an inner primer pair FIP / BIP and a loop primer pair LoopF / LoopB; the detection method is based on loop-mediated isothermal amplification (LAMP), the genotype is visually interpreted through color change of reaction liquid, a thermal cycler is not needed, and the detection method has the advantages of being high in accuracy and sensitivity, easy to operate, low in cost and the like. The matched kit comprises a rapid DNA extraction assembly and is suitable for field on-site detection. The gene mutation frequency of the specific site of the chilo suppressalis can be rapidly detected, and the technical support is provided for resistance monitoring, early warning and scientific treatment of bisamide insecticides.

This invention relates to a thermal unit for temperature management of a dPCR thermal cycler, and also to a thermal unit for a device for simultaneously thermally cycling multiple biological samples, also known as temperature cycling, such a device itself, and a method for simultaneously thermally cycling multiple biological samples using such a device and thermal unit. The thermal unit has at least one heat block (3) and a cooling structure (2), the heat block (3) comprising at least one thermoelectric transducer (31) and a heat transfer plate (33) attached to the thermoelectric transducer (31) for dissipating heat energy from the thermoelectric transducer (31), and the cooling structure (2) comprising a radiator (21), at least one fan (221), and exhaust pipes (24a, 24b).

The invention provides a method for providing skin analysis, and personalized skincare information based on that analysis, to a consumer at a retail point of sale, the method comprising: (i) at the retail point of sale, receiving a sample of the skin microbiome, which is collected from a skin surface of the consumer in a non-invasive manner, (ii) extracting the DNA from the sample of the skin microbiome and analysing the extracted DNA using a portable qPCR thermocycler which is configured to amplify and detect nucleic acids associated with a panel of microorganisms in the sample, (iii) uploading the analysis data from the thermocycler to a machine learning platform, and (iv) providing, at the retail point of sale, a profile of the consumer's skin microbiome and a personalized skincare routine for the consumer, generated from the analysis data using the machine learning platform.

The invention discloses a multiplex nucleic acid detection kit for swine acute diarrhea pathogens and application of the multiplex nucleic acid detection kit. The kit comprises an RAA (Reverse Amplification) primer pair, a specific crRNA (Ribonucleic Acid), a Cas12a protein, an NEBbuffer r2.1 and an ssDNA (Deoxyribose Nucleic Acid) reporter molecule, wherein the RAA primer pair, the specific crRNA, the Cas12a protein, the NEBbuffer r2.1 and the ssDNA reporter molecule are used for detecting the porcine epidemic diarrhea virus (PEDV), the porcine rotavirus (PoRV) and the porcine sapelovirus (PSV). According to the method, reverse transcription, amplification and Cas12a detection processes are coupled through an integrated system or a continuous reaction mode, the whole process is completed under the condition of constant temperature (37 DEG C), thermal cycle equipment is not needed, and only a constant-temperature heating device is needed. After the reaction is finished, a result can be observed through a portable blue light detector or naked eyes, so that on-site real-time visual interpretation is realized.

Systems and methods for processing samples in a multi-well reaction vessel can include inserting a multi-well reaction vessel into a heating chamber of a thermal cycler, enclosing the multi-well reaction vessel in the heating chamber, and compressing a bottom surface of the multi-well reaction vessel into a compliant thermally conductive insert to increase a thermal contact area between the bottom surface and the compliant thermally conductive insert. The compliant thermally conductive insert can be placed between the multi-well reaction vessel and a heating element of the thermal cycler, where heat flux from the heating element passes through the compliant thermally conductive insert to the reaction vessel. The compliant thermally conductive insert can include an elastically deformable creped graphite sheet that can reversibly deform according to different compression profiles depending on the topography or flexure of the reaction vessel and / or heating element.

The present invention relates to a device (10), system (200) and process for the remote calibration of a thermocycler (50).

The invention belongs to the technical field of molecular biological detection, and particularly relates to an RT-LAMP (Reverse Transcription Loop-Mediated Isothermal Amplification) and immunochromatography combined PEDV (Porcine Epidemic Diarrhea Virus) nucleic acid detection kit and application thereof. According to the kit, the high sensitivity and high specificity of the loop-mediated isothermal amplification technology and the rapid and visual interpretation advantages of the immunochromatography test strip are combined, and on-site, rapid and accurate detection of the PEDV is achieved. The core of the invention comprises a set of specific primer group designed for a PEDV N gene conserved region, an optimized RT-LAMP reaction system and an immunochromatography test strip which is specially treated and is used for detecting an LAMP amplification product. The kit does not need an expensive thermal cycler, the whole detection process is completed within 40 min, the sensitivity can reach 100 copies / [mu] L, and the kit is particularly suitable for rapid diagnosis and epidemic prevention and control of sites such as pig farms and grassroots veterinary stations.

A sample plate for a thermal cycler suitable for performing a polymerase chain reaction (PCR) procedure includes a base plate and a number of reaction vessels extending upward from the base plate. The sample plate further includes a vertical wall surrounding an outer perimeter defined by the reaction vessels. The vertical wall can be a continuation vertical wall, an intermittent vertical wall, or a perforated vertical wall. The intermittent vertical wall can include a plurality of wall portions, each of which plurality of wall portions is separated from other wall portions via a plurality of gaps.

The present invention relates to a thermal cycler having a thermal module of which the position is adjustable, the thermal cycler comprising: a thermal module for heating and cooling a reaction container; a first support structure to which the thermal module is mounted; a vertical guide part which guides the first support structure to be moved vertically; and a second support structure which supports the vertical guide part and accommodates the first support structure. Thus, a pressure between the reaction container and a heat lid can be uniformly distributed, thereby improving the lifespan of the heat lid and reducing misalignment between components. In addition, provided is the thermal cycler, in which the first support structure is configured such that the thermal module can be attachable / detachable, and the thermal module is attached / detached through one open side of the first support structure. Thus, in the aspect of maintenance, repair and replacement can be carried out easily, and specifications can be changed stably.

The present invention discloses a CRISPR-Cas12a system and detection method for visual detection of bovine viral diarrhea virus, which belongs to the field of virus detection technology. The present invention integrates reverse transcription cross primer amplification (RT-CPA) and the targeted cutting ability of CRISPR / Cas12a, combines RT-CPA pre-amplification with Cas12a signal amplification, and successfully constructs a bovine viral diarrhea virus detection platform, which significantly improves the sensitivity and specificity of BVDV detection. Compared with traditional RT-PCR, RT-CPA technology significantly simplifies the reaction conditions through isothermal amplification, avoids dependence on thermal cyclers, and improves amplification efficiency through cross primer design, so that the detection limit reaches 8.0 copies / μL.

The application provides a thermal cycle experiment device, and relates to the field of experimental equipment. The thermal cycle experiment device comprises a heating furnace, an execution unit and a regulation unit. The heating furnace has a furnace cavity for accommodating a sample and an inlet and outlet connected with the furnace cavity. The execution unit comprises a sample disc, a connecting rod and a cover, the sample disc and the cover are connected with the connecting rod, the sample disc is used for carrying the sample, and the cover is used for opening or closing the inlet and outlet. The regulation unit is connected with the connecting rod and is used for driving the connecting rod to slide along the extension direction of the connecting rod, so that the sample disc enters or leaves the furnace cavity; and after the sample disc enters the furnace cavity, the cover closes the inlet and outlet. The thermal cycle experiment device does not need to realize the thermal cycle simulation of the sample by temperature rising and falling, improves the influence of the temperature change process on the experimental result, and is more flexible and reliable in temperature control.

The invention provides RPA (recombinase polymerase amplification) detection primers for colletotrichum gloeosporioides of green vegetables and application of the RPA detection primers and belongs to the technical field of agriculture. In order to solve the problems of long time consumption (2-3 hours), low sensitivity (5 ng / mu L) and dependence on precise equipment in the existing PCR detection, the invention provides RPA primers 56F1 (5 '-GAAAAGCGGCAT-3') and 56R1 (5 '-CGTTGGAAAATACA-3') of targeted pathogenic bacteria specific genes NC030956, as well as an RPA primer 62F1 / F1 and an RPA primer 62F2 / F2, and establishes a 39 DEG C constant-temperature 20-minute amplification system. The method realizes three major breakthroughs: 1, the detection sensitivity reaches 0.05 ng / mu L and is improved by 100 times compared with PCR (Polymerase Chain Reaction); 2, seven related pathogens such as colletotrichum nicotianae are strictly distinguished, zero cross reaction is realized, and the sample accuracy rate is 100%; 3, the whole process only needs 1.5 hours, the DNA crude extraction time of diseased leaves is 30 minutes, the RPA amplification time is 20 minutes, the electrophoresis detection time is 35 minutes, the cost is low, and a thermal cycler is not needed. The invention provides an on-site solution with the characteristics of rapidness, accuracy and saving for prevention and control of the cruciferous anthracnose, and solves the problem that the traditional technology cannot give consideration to both speed and sensitivity.

The application discloses a mycoplasma pneumoniae 23S rRNA gene A2063G drug resistance mutation detection method based on a CRISPR-LbCas12a system and belongs to the technical field of molecular diagnosis. The detection method can accurately distinguish single base differences such as A2063 and G2063, effectively avoids mismatch, and ensures the accuracy of results. Relying on a one-step reaction of RPA and CRISPR, the total time consumption is less than 45 minutes, and the waiting time is significantly shortened. The whole reaction is carried out under constant temperature conditions, does not need to rely on complex equipment such as a thermal cycler, is easy to operate, is very suitable for the application of primary medical institutions and instant test scenes. The cost control advantage is outstanding, the reagent cost is about 1 / 50 of second-generation sequencing and 1 / 5 of fluorescent PCR, and the economy is extremely good. In addition, the application technology shows good multiplex detection potential and wide application prospect.

The present invention provides a thermal cycler capable of rapidly and efficiently heating and cooling a reaction liquid. The thermal cycler according to the present invention comprises: a temperature control block where a reaction vessel can be installed, a thermoelectric conversion unit capable of heating and cooling, a temperature sensor that measures the temperature of the temperature control block, an insulating substrate that is in contact at one surface with the thermoelectric conversion unit, and a heat radiating unit that is provided on the other surface of the insulating substrate and serves for discharging the heat of the thermoelectric conversion unit to the outside, wherein the temperature control block is heated and cooled by controlling a current or voltage supplied to the thermoelectric conversion unit on the basis of the temperature of the temperature adjustment block measured by the temperature sensor.

A thermal cycler system comprises a sample block configured to receive a sample holder configured to receive a plurality of samples; an adaptor configured to surround a periphery of the sample block; and a drip pan configured to surround a periphery of the adaptor. The drip pan comprises one or more ejector mechanisms and one or more openings, the one or more ejector mechanisms configured to respectively extend through the one or more openings into contact with the sample holder in a state of the sample holder received by the sample block and the adaptor surrounding the periphery of the sample block.

The invention discloses a primer and a probe for real-time RPA (recombinase polymerase amplification) detection of goose parvovirus and a detection method. The primer comprises an upstream primer G-F1 and a downstream primer G-R1, the sequences of the upstream primer G-F1 and the downstream primer G-R1 are as shown in SEQ ID No.1-2, the probe is an Exo probe, and the sequence of the Exo probe is as shown in SEQ ID No.3. The detection limit of the detection method established by using the primer and the probe is as low as 3.0 * 10 < 1 > copy number / mu L, cross reaction with other waterfowl viruses is avoided, the clinical coincidence rate reaches 98.1%, detection is completed within 30 min in the whole process, a thermal cycler is not needed, and the method is suitable for field application. The matched kit comprises a primer pair, a probe and an RPA basic reagent, is simple and convenient to operate and controllable in cost, can be used for rapid diagnosis and epidemiological monitoring of GPV classical strains and variants (MDGPV and SBDSV), and has important application value.

The invention relates to the technical field of biosensors, and discloses a p53 gene detection signal amplifier, a kit and a method for detecting a p53 gene in a sample. The signal amplifier comprises a hairpin probe HP and a lock-type probe, wherein the hairpin probe HP comprises an HP-1 segment which is complementarily paired with the p53DNA segment and an HP-2 segment which is complementarily paired with the lock-type probe. The signal amplifier provided by the invention has the advantages of high specificity, high sensitivity and the like, so that the method provided by the invention can be used for rapidly, efficiently and accurately detecting the p53 gene in a sample. Moreover, the signal amplifier and the method provided by the invention do not depend on high-price instruments such as a high-quality thermal cycler, and are very suitable for popularization and application in grassroots.

The invention relates to a thermoblock for receiving and regulating the temperature of at least one laboratory specimen vessel in a laboratory temperature regulation device, more particularly in a PCR thermocycler, wherein the thermoblock has been produced by means of an additive manufacturing method using a material containing metal. The invention further relates to a method for producing the thermoblock and a computer-implemented method for the simulation of physical characteristics of the thermoblock to be produced in accordance with the invention.

The invention provides a digital PCR thermal cycler. The digital PCR thermal cycler comprises a shell; a door opening and closing module (12); the thermal cycler comprises a shell (1), a temperature control module (13), and an air pump (17), an air storage tank (16), a separator (18) and a water storage module (19) which are arranged in the shell, the air pump (17) is communicated with the separator (18) through a first air supply pipeline (44), the separator (18) is used for removing water in air, and the separator (18) is communicated with the air storage tank (16) through a second air supply pipeline (45); wherein the water storage module (19) is in fluid communication with the separator (18) to receive water separated from the air by the separator (18). According to the digital PCR thermal cycler, impurities such as oil and water in compressed air can be removed, and moisture in the air is prevented from entering systems such as an air storage tank and an air passage; improved door opening and closing operation is provided, and the sealing performance of a sealed pressure maintaining environment is improved; and moreover, the three areas independently control the temperature, so that the heat uniformity is good.

The invention relates to a kit for detecting nocardia seriolae of jewfish based on RPA-CRISPR / Cas12a as well as a detection method and application of the kit. The main component of the kit is a composition for identifying nocardia seriolae, and the composition comprises a specific primer for detecting nocardia seriolae based on RPA-CRISPR / Cas12a, a Cas12a protein, crRNA and a fluorescence reporter probe ssDNA Reporter, wherein the specific primer, the Cas12a protein, the crRNA and the fluorescence reporter probe are used for detecting nocardia seriolae based on RPA-CRISPR / Cas12a. The detection method of the kit comprises the following steps: S1, extracting a sample genome DNA; s2, carrying out RPA (recombinase polymerase amplification); s3, CRISPR (clustered regularly interspaced short palindromic repeats) / And S4, interpreting a detection result. According to the invention, a target gene segment of nocardia seriolae is rapidly amplified at a constant temperature by virtue of an RPA technology, and the target gene segment of nocardia seriolae is specifically targeted by virtue of accurate recognition and trans-cleavage capabilities of CRISPR / Cas12a, so that rapid detection visible to naked eyes is realized. Compared with conventional detection methods such as PCR, qPCR and the like, the kit does not need a professional thermal cycler, has the advantages of high sensitivity, strong specificity, simplicity and convenience in operation, low cost and the like, and is expected to be applied to on-site instant detection of aquaculture sites and basic laboratories.

The invention provides a kit for detecting jewfish aeromonas veronii based on RPA-CRISPR / Cas12a, a detection method and application of the kit. The main component of the kit is a composition for identifying the aeromonas veronii, and the composition comprises a specific primer for detecting the gyrB gene of the aeromonas veronii based on RPA-CRISPR / Cas12a, a Cas12a protein, crRNA and a fluorescence report probe ssDNA Reporter. The detection method of the kit comprises the following steps: S1, extracting a sample genome DNA; s2, carrying out RPA (recombinase polymerase amplification); s3, CRISPR (clustered regularly interspaced short palindromic repeats) / And S4, interpreting a detection result. According to the invention, a target gene segment of the aeromonas veronii is rapidly amplified at a constant temperature by virtue of an RPA technology, and the target gene segment of the aeromonas veronii is specifically targeted by virtue of accurate recognition and trans-cleavage capabilities of CRISPR / Cas12a, so that rapid detection visible to naked eyes is realized. Compared with conventional detection methods such as PCR, qPCR and the like, the kit does not need a professional thermal cycler, has the advantages of high sensitivity, strong specificity, simplicity and convenience in operation, low cost and the like, and is expected to be applied to on-site instant detection of aquaculture sites and basic laboratories.

The invention discloses a miniaturized portable PCR-SERS nucleic acid instant detection platform, and belongs to the technical field of nucleic acid detection. The platform is composed of a miniaturized thermal cycler, a miniature PCR reaction cavity, a filling type detection pen and a lateral flow chromatography test strip, a core component of the miniaturized thermal cycler is an inner core capable of providing temperature required by PCR amplification reaction, and the miniature PCR reaction cavity is composed of an upper-layer heat insulation adhesive tape, a PCR amplification paper film and a lower-layer heat insulation adhesive tape. The PCR amplification paper membrane is composed of a cellulose acetate membrane and a reagent which is freeze-dried on the cellulose acetate membrane in advance and is required by a PCR reaction, and a surface carboxylation SERS probe which is activated by EDC / NHS and is coupled with targeted recognition molecules is sprayed on a combination pad of the lateral flow chromatography test strip. The miniature thermal cycler has the beneficial effects that the volume of the miniature thermal cycler is about 35cm < 3 >, so that the miniature thermal cycler is very convenient to carry; reagents required by the PCR reaction are freeze-dried on the cellulose acetate membrane in advance, so that reagent-free operation in the whole process is realized.

The present invention relates to a portable laboratory for detecting phytopathogenic microorganisms using a plant sample, comprising at least the following elements: a thermocycler suitable for qPCR; a minicentrifuge; a scale; a manual homogeniser suitable for breaking the plant sample, and a kit for extracting and detecting DNA specific to pathogens of plant species. The manual homogeniser comprises a handgrip for holding a shaft terminating in a surface with bearings for breaking the plant sample and the thermocycler suitable for qPCR has the ability to analyse up to 16 samples simultaneously.

A reaction vessel comprises a lower chamber with a first volume, and an upper chamber with a second volume greater than the first volume. A thermocycling system for heating the reaction vessel includes a lower heating zone to heat the lower chamber, an upper heating zone to heat the upper chamber, and a lid heater to heat an opening of the upper chamber. A method comprises loading a sample into a lower chamber of a reaction vessel, thermocycling the lower chamber using a lower heating zone of the thermo cycler, combining an additive into the sample to produce a combination filling the lower chamber and at least partially filling an upper chamber of the reaction vessel, and incubating the upper and lower chambers using the lower heating zone and an upper heating zone. The lower and upper chambers can have different wall thicknesses to facilitate heat transfer.

The invention relates to the technical field of molecular detection, and particularly discloses an isothermal amplification detection marker, probe, kit and detection method for identifying robinia pseudoacacia honey, and the method comprises the following steps: (1) extracting genome DNA in a honey sample to be detected; (2) the DNA obtained in the step (1) is used as a template, an isothermal amplification reaction system is used for amplification, and amplified target nucleic acid is a robinia pseudoacacia species specific DNA fragment; and (3) carrying out contact reaction on the amplification product in the step (2) and a lateral flow chromatography test strip, and judging whether the honey sample contains the robinia pseudoacacia component or not. The isothermal amplification technology based on the robinia pseudoacacia specific DNA fragment is combined with lateral flow chromatography detection, so that the accuracy and high sensitivity of a detection result are ensured, related species can be effectively distinguished, and trace target components can be detected; the characteristics of constant temperature and rapidness and the visual result interpretation mode get rid of dependence on a precise thermal cycler, and the operation difficulty, the time cost and the equipment threshold are greatly reduced.

An automated method for performing an amplification reaction includes the automated steps of combining a fluid sample together with one or more amplification reaction reagents in a reaction receptacle using a first automated pipettor, thereby forming a reaction mixture, transporting the reaction receptacle to a first location of a centrifuge, subjecting the reaction mixture to centrifugation, after subjecting the reaction mixture to centrifugation, removing the reaction receptacle from a second location of the centrifuge different from the first location and placing the reaction receptacle in a thermal cycler, and in the thermal cycler, subjecting the reaction mixture to one or more temperature cycles.

The present invention relates to a composite material for a thermal block of a thermal cycler. The composite material has both low specific heat characteristics and remarkably improved thermal conductivity, and thus, when the temperature of a thermal block, which is manufactured using the composite material, rises and falls, deviation in temperature change rate between a plurality of unit thermal blocks and / or in different areas in a unit thermal block can be remarkably reduced such that a PCR thermal cycler including the thermal block can ensure reliability even in ultrafast PCR.

The invention discloses a colorimetric detection method and a colorimetric detection kit for staphylococcus aureus based on CRISPR (clustered regularly interspaced short palindromic repeats) / Cas12a-DNAzyme. According to the colorimetric detection method and the colorimetric detection kit, a specific nuc gene (an NCBI (National Center of Biotechnology Information) accession number V01281.1) of the staphylococcus aureus is taken as a target. Carrying out centrifugal impurity removal and boiling water cracking on the sample to obtain a nucleic acid template, and carrying out isothermal amplification on the nuc gene through RAA; cas12a protein is guided by targeting crRNA to recognize an amplification product, a trans-cleavage active cleavage CatG4R probe is activated, released CatG4 and hemin form DNAzyme, and ABTS-H2O2 is catalyzed to develop color. The detection limit is 100 CFU / mL, and the time consumed in the whole process is less than 2 hours. The specificity of the method is far better than that of an existing target; the pretreatment is simplified to be within 20 minutes, and an expensive kit is not needed; rAA does not need a thermal cycler, color comparison realizes naked eye interpretation, and equipment dependence is greatly reduced. The method is suitable for dairy product detection, and solves the problems of slow speed, poor immunoassay specificity and the like of a traditional method.