170 results about "Pcr chip" patented technology

Provided is a device for injecting a PCR solution into PCR channels of a PCR chip. The device includes: first through-holes corresponding to each of the inlets of the PCR chip in which PCR channels are formed; and second through-holes corresponding to each of the outlets of the PCR chip. The first through-holes are aligned with each of the corresponding inlets of the PCR chip and the second through-holes are aligned with each of the corresponding outlets of the PCR chip when the PCR solution is injected so that the PCR solution can flow into the inside of the PCR channels through the first through-holes or the second through-holes. The device slides to seal the inlets and the outlets of the PCR chip after the PCR solution is injected.

The portable micro PCR chip system prepared by MEMS or micro-model technique in bio-micro EMC system field comprises some trap-type reaction ponds for DNA amplification connected to liquid inlet/outlet by flow passages, an integrated micro-heater and sensor for amplifying DNA by the temperature cycle control system, a micro-pump and micro-valve on pond inlet to lead the DNA initiator and ponds insulation, and maybe a fluorescence excitation source, a detector and a fluorescence signal detection and analysis system for on-line detection on PCR product. This chip system provides a high-flux and fast DNA amplification means with low cost and well reliability.

The invention relates to a method for manufacturing a digital PCR (polymerase chain reaction) chip based on a mineral-oil saturated PDMS (polydimethylsiloxane) material. The method is characterized in that the digital PCR chip based on PDMS is prepared from a PDMS monomer of a certain amount of mineral oil (liquid paraffin) and comprises an emulsion droplet generation structure and an emulsion droplet collection structure. After the emulsion droplets are made and collected on the same chip, the emulsion droplets are subjected to PCR amplification on the same chip. The phagocytosis to the oil phase in the digital PCR system by the PDMS of the chip can be avoided, the emulsion droplets can be kept stable during PCR, and the stability of the PCR can be guaranteed. In addition, compared with the existing technology of the digital PCR chip, the method provided by the invention is low in cost, is convenient to operate and has a very wide application prospect.

Provided are a polymerase chain reaction (PCR) module and a PCR system including the same. The PCR module includes: a detachable PCR chip including a PCR chamber unit in which a PCR solution is accommodated; a heater unit for heating the PCR solution in the PCR chip with a preset temperature; a detecting unit for detecting a PCR signal of the PCR solution; a PCR chip installation unit for mounting/detaching the PCR chip using a one-touch method, in which the heater unit is adhered to the PCR chip with a predetermined pressure when mounting the PCR chip and the heater unit is separated from the PCR chip when detaching the PCR chip; and a housing covering at least the heater unit and the detecting unit so that they are not exposed to the outside.

The invention discloses a large-area digital PCR microsphere fluorescence high-flux detection device, which comprises a fluorescence exciting light path and a fluorescence detection light path, wherein along the fluorescence exciting light path, exciting light emitted by a light source device sequentially passes through a focusing device and an objective lens device to be emitted onto a digital PCR chip of an objective table; along the fluorescence detection light path, the fluorescence generated by the digital PCR chip being excited by the exciting light sequentially passes through the objective lens device, the focusing device and an imaging device to be imaged and is then transmitted to an image collection sensor; the objective table is also provided with a temperature control device; the temperature control device controls the circulation temperature of the reaction of the digital PCR chip in real time. The invention discloses a large-area digital PCR microsphere fluorescence high-flux detection method. The higher detection efficiency and the large-area high-flux detection accuracy can be obtained. The method and the device can realize the real-time digital qPCR detection with high flux, high sensitivity and high specificity; the important significance is realized on improving the performance, the precision and the flux of a digital PCR detection instrument.

The invention discloses a PCR (Polymerase Chain Reaction) chip based on droplet array. The chip is obtained by taking a mono crystalline silicon sheet or a glass sheet as a base material and adopting standard photoetching as well as wet etching technique. The mono crystalline silicon sheet or the glass sheet is the base material of the chip. A SiO2 oxide layer and a silane layer generated by silanization on the surface of the SiO2 oxide layer are adhered to the base material successively. A hydrophilic pit array, which can be a droplet array and is formed by etching with photoresist technology, is disposed on the silane layer. A closed circular guardrail is arranged on the outer periphery of the hydrophilic pit array and the guardrail is connected with the silane layer in a sealed manner. The invention also discloses the application of the chip on real-time quantitative PCR detection and real-time quantitative isothermal amplification reaction detection. The detection system provided in the invention remarkably reduces the reaction volume and the detection sensitivity of the system fairly equals that of a commercialized quantitative PCR instrument. The detection system in the invention is easy for operation and has low cost, thus can be applied to commercialization.

The invention discloses a micro fluidic chip apparatus by integrating a continuous flow PCR and a capillary electrophoresis function, which comprises a continuous flow type PCR chip and a capillary electrophoresis chip, and is characterized in that an output channel of the continuous flow type PCR chip is directly connected in a sample pool of the capillary electrophoresis chip, a sample introduction channel and a separating channel are provided on the capillary electrophoresis chip, the sample introduction channel and the separating channel present an intersect shape, the top of the sample introduction channel is the sample pool and a sample waste liquid pool, and the top of the separating channel is a buffer liquid pool and a waste liquid pool. The apparatus provided by the invention can enhance the automation degree and whole process operation speed for DNA detection, the operation step is reduced, and the apparatus is convenient for miniaturization and portable performance of the apparatus.

The invention provides a micro-droplet type PCR (polymerase chain reaction) chip and a manufacture method thereof. The upper layer of the chip is a PDMS (polydimethylsiloxane) imaging cover plate formed by connecting a DNA (deoxyribonucleic acid) target fragment with a primer injection port, a single phase drop forming microstructure, a PCR reaction enzyme and an mRNA (messenger ribonucleic acid) substrate injection port, an oil phase reagent injection port, a drop straight line channel, a cross hybrid channel, a micro drop buffer channel, a micro drop PCR reaction chamber and a micro drop PCR waste liquid chamber, the lower layer of the chip is a glass substrate, and the upper layer and the lower layer are bonded and packaged to form a complete micro drop PCR chip. The manufacture method of the chip comprises the steps of firstly, manufacturing a mask plate; secondly, molding; thirdly, pouring a die; and fourthly, taking a figure. The PDMS figure at the upper layer of the chip adopts a multilayer filter structure, and the formed micro drop has a uniform size, stable grain diameter and strong controllability. Compared with traditional PCR chips, the micro drop PCR chip manufactured by the method has great advantages in hybrid effect, reaction efficiency, and high throughput.

The invention discloses automatic reaction device and method for pretreating cells. The device comprises a reaction box body, a base, a box cover and a piston, wherein a middle cavity is formed in the center of the reaction box body; seven cavities are formed in the periphery of the hollow cavity; seven square cavities are correspondingly formed between the middle cavity and the seven cavities in the periphery; a control valve is put into each square cavity; communication between an inner chamber and an outer chamber is controlled through up and down movement; the lower part of the reaction box body is tightly connected with the base; the box cover is arranged at the upper part of the reaction box body; a piston rod which is matched with the middle cavity penetrates through the box cover; and the outer side of a second PCR reagent cavity is connected with a PCR chip for detecting nucleic acid. According to the reaction device and method, crushing and cracking of original samples, and purifying and extracting of the nucleic acid and PCR amplification are integrated in a reaction box; the operation process is simplified; the pretreatment efficiency is greatly improved; meanwhile, the reaction volume is reduced; the portability of an instrument is improved; and a foundation is laid for rapid, safe and accurate nucleic acid detection.

The invention provides a digital PCR chip and a using method thereof. The chip comprises a silicon microporous chip, a microfluidic channel for injecting a sample and an aluminum heat conduction base, wherein 20,000 through holes with hydrophilic inner walls and hydrophobic surfaces are processed on the silicon microporous chip. A hydrophilic nucleic acid or cell sample, when flowing through the part above the silicon microporous chip, is spread, monodispersed and retained in each micropore, and mineral oil is introduced, so that the whole space beside the silicon microporous chip is filled, and complete isolation of each microreaction system is realized. The digital PCR chip has the advantages of novel design, easiness in use, low cost and wide application range, and compared with the conventional method for monodispersing the nucleic acid sample in a water-in-oil process, is lowered in cost by more than 10 times, is short in sample preparation time and simple in operation method, is excellent in accuracy of dispersing the nucleic acid sample and reliability of the reaction system, and is small in cross pollution.

The invention provides a digital PCR (polymerase chain reaction) chip. The digital PCR chip comprises a droplet production module formed by laminating a cover glass and a glass substrate and a droplet collecting module tightly attached to one surface of the droplet production module, wherein two through holes including a liquid phase sample inlet and an oil phase sample inlet are formed in the cover glass; microflow channels communicated withthe liquid phase sample inlet or the oil phase sample inlet are etched in one surface, attached to the glass substrate, of the cover glass, each microflow channel is divided into two branch channels, and the branch channels pairwise intersect with droplet production ports; the droplet collecting module comprises a cavity with two opening sides, and the droplet production ports are all tightly attached to the inner side of one opening. The digital PCR chip is provided with the independent droplet collecting module, the PCR and the result observation are both performed in the chip, so that the probabilities of pollution and demulsificationare greatly reduced, meanwhile, results of the PCR are observed under a fluorescence microscope in a unified manner, signals reading of droplets is not required to be performed in the chip one by one, and the signal reading time is greatly shortened.

The invention relates to a multiple digital PCR chip and a use method thereof. The chip does not need to be driven by a precise micro-pump and be controlled by a complex micro-valve and does not need a complex macro-micro interface. The chip realizes simple, fast, low cost and large-scale autodecomposition and quantitative uniform distribution of samples. Compared with the existing commercial PCR chip, the chip greatly reduces the complexity and operating cost of the digital PCR system, simplifies the experimental operation and is free of professional training of an operator. The chip realizes the independence of each PCR reaction unit through microcavity arrays, is free of a surfactant and prevents the influence caused by an additive on the reaction system. Through integration of the isolated microcavity array zones and pre-deposition of primers corresponding to different specific target molecules in the microcavities in different zones, the chip can realize multiple detection through one step. The chip has the advantages of operation simpleness, low cost and strong multiple detection capacity and can promote development and wide application of the digital PCR technology.

The present invention discloses a kind of control device and method for PCR chip array. The PCR chip array without built-in heater and sensor is set on a heat exchanger; four liquid storages store liquid of different temperature; and circulating pumps and eight solenoid valves make liquid of proper temperature to circulate inside the heat exchanger for the chips get required reaction temperature. Over the chip array, is the flexible printed circuit board with heaters and sensors corresponding to the chips for the dynamic trimming of the temperature of the chips. The upper surface of the heat exchanger is divided into several temperature zones, and each of the temperature zones has one chip selected as the template chip with built-in sensor and filled liquid and with the measured temperature value used as the feedback signal for controlling the temperature of the chips in the said zone. One computer with interface circuit is used for the feedforward-cascade control and statistics process.

The invention discloses a method for manufacturing a digital PCR (polymerase chain reaction) chip based on a 3D (three-dimensional) printing platform. The method comprises the steps as follows: step one, a substrate with a micropore array structure is adopted; step two, the substrate is subjected to surface treatment; step three, the substrate is placed on the 3D printing platform; step four, by means of a plurality of nozzles on the 3D printing platform, the substrate is subjected to quantitive jet printing in oil drop and liquid drop manners alternately, so that a liquid drop in oil array structure is formed in a micropore on the surface of the substrate; step five, the micropore array structure on the substrate is filled with excess oil; step six, a transparent cover sheet is adopted; step seven, the transparent cover sheet is fixed on the substrate for covering, so as to finish the preparation. According to the method for manufacturing the PCR chip based on the 3D printing platform, a 3D printing technology is adopted to build a picoliter magnitude reaction system and achieve an amplified reaction in the system, therefore, the efficiency is high, the cost is low, and the maximum possibility is provided for the digital PCR productization.

The invention relates to a microfluidic PCR (Polymerase Chain Reaction) chip fluorescence fluid detection device based on a CCD (Charge Coupled Device) image sensor. The device is composed of a spatial three-dimensional micro-channel, a CCD image sensor, a monochromatic LED (Light Emitting Diode) light source, a microprocessor and a liquid crystal display device, and the device is mainly used for carrying out fluorescence detection of a PCR amplification reaction in a spatial range and integrating signal generation, signal collection and transmission and signal processing and recognition parts in signal detection to achieve integration, automatic detection and control of a miniature microfluidic real-time fluorescence PCR detection system, thereby avoiding optical path transmission of an optical path composed of various optical devices and optical fibers, simplifying the detection structure and facilitating the integration of the system.

The invention relates to a chip structure for PCR (polymerase chain reaction) rapid reaction. The chip structure for PCR rapid reaction comprises a cover plate, a medium layer, a PCR chip and an external heating structure; through holes are machined on the cover plate; micro cavity holes are arranged on the PCR chip; the medium layer is covered on the PCR chip, and absorbs the cover plate in a negative pressure mode through the through holes on the cover plate; the lower bottom surface of the PCR chip are in tight contact with the external heating structure; the PCR chip can be a single-layer structure, and the micro cavity holes are directly machined; or the PCR chip can be a multi-layer structure, and micro cavity holes are machined on at least one layer, and are then combined with the other base material. PCR amplification is easy to achieve rapidly, and the biochemical reaction speed is improved obviously; the heat capacity of a micro environment of a chip is small, and temperature rising and reduction can be controlled at high speed, and the regional temperature can be detected and controlled accurately; the temperature cycle system volume is reduced, the heat capacity is reduced, and the reaction time is shortened greatly; the reaction liquid volume is reduced, the amplification specificity is intensified, and not only is cost saved, but also the efficiency is improved.

The present invention is the electrochemical preparation process of quantitative PCR detection chip, and relates to the electrochemical detection technology for quantitative nucleic acid PCR chip. The preparation process includes preparing a micro electrode array on the surface of solid carrier; fixing nucleic acid capturing molecular probes on the electrodes of the micro electrode array; preparing closed micro cavity for carrying liquid in the electrode area of molecular probes; and connecting wires to the electrodes on the solid carrier. The detection process includes adding nucleic acid, enzyme, deoxyribonucleotide, electrochemically active matter and other reaction components into the micro cavity; setting the chip in large cavity for controlling temperature; detecting the current and voltage variance caused by reaction of the captured DNA probe molecule with constant potential instrument; and detecting the variance rule of PGR circulating process in different electrodes for PCR detection of genes.

The invention provides a digital PCR chip signal reading method. The method comprises the following steps: adding a reaction solution in several reaction chips; performing amplification on each reaction chip; obtaining a fluorescence reagent image of each reaction chip; performing normalization treatment on each fluorescence reagent image; reading the effective aperture numbers in each reaction chip to obtain the total effective aperture number; reading the fluorescence intensity in apertures in each reaction chip, establishing a fluorescence histogram; according to the fluorescence histogram, setting the positive determination threshold of each reaction chip to obtain the total positive locus number, according to the ratio of the total positive locus number to the total effective aperture number, and calculating the concentration of a to-be-measured sample solution. The amplification system and the detection system is completely compatible with a current micropore chip-type digital PCR technology, several chip fluoroscopic images after amplification are obtained through a multitime imaging mode, the histogram and point diagram analysis can be carried out on the amplified chip image to obtain the digital PCR detection result with high flux, and the detection precision is increased.

The invention discloses a glass capillary minitype polymerase chain reaction (PCR) system and a preparation method thereof. The PCR system comprises a chip base, a PCR chip and a fan. The PCR chip is composed of glass capillary and a bottom plate. The glass capillary is internally provided with a PCR reaction chamber. A heating electrode is arranged on the lower surface of the bottom plate, a temperature measuring electrode and a deep groove are formed in the upper surface of the bottom plate, the glass capillary is embedded into the deep groove, and the deep groove is filled with heat conduction resin. The PCR chip reaction chamber is the glass capillary and can be used for achieving the PCR chip which is quick in temperature rising and reducing and temperature uniformity and high in amplification efficiency. The consumable item in the glass capillary minitype PCR chip is the glass capillary, the bottom plate with the heating electrode, the temperature measuring electrode and the deep groove can be used for multiple times, cross contamination is avoided, and the use cost of the chip is greatly reduced.

Aspects of the present disclosure relate to DNA amplification systems, e.g., PCR chips used to hold a testing solution and sample, and systems and methods for testing the sample including thermocycling and detection. The system can include a containment chip capable of holding a sample and reagents for amplifying a nucleic acid in the sample and containing a unique identifier used to choose a nucleic acid amplification protocol. The system can also include a nucleic acid detection component which may contain a light source, a light sensor, a temperature control unit and a processor.

The invention belongs to the technical field of virus in-vitro nucleic acid detection and particularly relates to a digital PCR quantitative detecting kit for HBV (hepatitis B virus) cccDNA and an application of the digital PCR quantitative detecting kit for HBV cccDNA. A digital PCR quantitative detecting probe composition of the HBV cccDNA is firstly designed, and the kit comprises a container for detecting the probe composition and a container for a purifying reagent containing liver puncture tissue DNA. Using of the kit comprise the following steps of extracting the DNA in the liver puncture tissue, and adopting PSAD (prostate specific antigen density) enzyme to digest the HBV cccDNA; by taking a digital PCR detecting chip as a reaction container and adopting a complex fluorescent multiple PCR primer, amplifying target DNA in detecting micro pores of the digital PCR chip; and obtaining the quantitative results of the HBV cccDNA in each cell according to a fluorescence signal in each micro pore of the digital PCR detecting chip. The digital PCR quantitative detecting kit for HBV cccDNA is strong in specificity, high in sensitivity, simple and convenient and quick. HBV cccDNA and HBV rcDNA are not needed to be separated and extracted, and the defects in the conventional HBV cccDNA detection are overcome, and therefore, the digital PCR quantitative detecting kit is suitable for large-scale popularization and application.

The invention discloses a liquid drop capturing chip. The liquid drop capturing chip comprises a plurality of fluid micro channels arranged in parallel, wherein a plurality of liquid drop capturing structures are formed along the fluid micro channels at intervals; the liquid drop capturing structures comprise liquid drop entering ends and inhibition liquid drop flowing-out ends; and the inhibition liquid drop flowing-out ends are communicated with the adjacent fluid micro channels. The invention further discloses a fully integrated digital PCR chip based on a seaweed gel liquid drop and application thereof. The digital PCR chip based on the seaweed gel liquid drop, disclosed by the invention, can finish the full process from the generation of liquid drops to capture of the liquid drops and to the detection of the liquid drops. And moreover, seaweed gels are introduced; after the seaweed gels are solidified, the seaweed gel liquid drops are stable, cannot be fused and cannot affect the PCR amplification efficiency; and the method can be applied to the detection of circulating tumor DNA in the blood and other related biology and medical sciences; and due to the introduction of the seaweed gels, the application can be further expanded to single cell analysis and so on.

An intelligent parallel detection instrument for the hybridization of PCR chips is composed of temp cycling unit, hybridizing unit, 3D transfer manipulator, detecting unit and computer. Said detecting unit has position sensor and photoelectric converter. Its detection process includes such steps as introducing PCR chip to temp controlled region, performing PCR procedure, moving PCR chip to hybridizing region, performing hybridizing procedure, washing, testing and processing the tested result.