WO2018028447A1 - Appareil, procédé et système pour la mise en oeuvre d'une réaction chimique - Google Patents

Appareil, procédé et système pour la mise en oeuvre d'une réaction chimique Download PDF

Info

Publication number
WO2018028447A1
WO2018028447A1 PCT/CN2017/095000 CN2017095000W WO2018028447A1 WO 2018028447 A1 WO2018028447 A1 WO 2018028447A1 CN 2017095000 W CN2017095000 W CN 2017095000W WO 2018028447 A1 WO2018028447 A1 WO 2018028447A1
Authority
WO
WIPO (PCT)
Prior art keywords
reaction
thermostat
module
receiving unit
reaction mixture
Prior art date
Application number
PCT/CN2017/095000
Other languages
English (en)
Chinese (zh)
Inventor
刘树谟
Original Assignee
皮卡（上海）生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 皮卡（上海）生物科技有限公司 filed Critical 皮卡（上海）生物科技有限公司
Publication of WO2018028447A1 publication Critical patent/WO2018028447A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38—Temperature-responsive control
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • B01L7/5255—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones by moving sample containers

Definitions

  • the present application relates to apparatus, methods, systems, and computer readable media for performing chemical reactions, particularly apparatus, methods, systems, and computers that utilize thermal cycling for sample detection and analysis (eg, for nucleic acid amplification). Read the media.
  • Nucleic acid amplification methods allow selective replication of nucleic acid sequences, thereby allowing selective enrichment and identification of target nucleic acids from complex mixtures, such as biological samples.
  • the biological sample is first treated to separate the nucleic acid from other components in the biological sample and other materials that may interfere with nucleic acid amplification.
  • the target nucleic acid can be amplified by amplification methods known in the art, such as thermal cycle-based polymerase chain reaction (PCR). After amplification of the target nucleic acid, the amplification product can be detected and interpreted by the end user.
  • PCR polymerase chain reaction
  • Bedside detection or on-the-fly detection has the ability to provide test reports quickly and accurately in situations where laboratory infrastructure is poor, resources are limited, or where real-time requirements for test results are high.
  • POCT also enables existing levels of health care facilities to be more likely to provide immediate feedback to patients during a single visit.
  • the inefficiency of POCT methods and devices limits the effects that can be achieved. For example, preparing nucleic acids (eg, nucleic acids from pathogens) from complex samples (eg, biological samples) requires skilled technicians in specialized laboratories. Multiple processing steps and subsequent tests are performed, and reports and results are often produced after hours or even days.
  • the speed of nucleic acid amplification devices and methods is still insufficient, or the speed is slow (for example, based on thermocouple heating and cooling), or the efficiency and sensitivity are low (various constant temperature amplification reactions), or the body size is large (for example, mechanical transfer reaction tubes) Inserted at different temperatures of the thermal module), or high cost, low reaction mixture capacity, and poor versatility (for example, various reaction tubes using microfluidic components or heaters). Therefore, devices and methods that are fast, efficient, portable, low in energy consumption, and low in manufacturing and use, particularly nucleic acid amplification methods, are urgently needed in a related field and society as a whole.
  • the present application provides devices, methods, and systems for performing chemical reactions, particularly nucleic acid amplification, such as polymerase chain reaction, to enable immediate detection of a sample.
  • a chemical reaction for example, a biochemical or molecular biological reaction
  • PCR polymerase chain reaction
  • the apparatus, methods, and systems provided herein enable rapid, low power, portable, easy to operate, and/or accurate results.
  • the devices, methods, and systems provided herein do not require a specially tailored reaction vessel and conventional PCR reaction tubes can be used.
  • the application provides a device for performing a chemical reaction that requires thermal cycling in at least two temperature intervals.
  • the apparatus can include a sample receiving unit rotatable about at least one axis of rotation, wherein the sample receiving unit is mated and thermally coupled to a reaction vessel for performing the chemical reaction, the sample receiving unit including at least two thermostats a module, and the at least two thermostat modules are maintained in different temperature zones from each other; and a control unit, the control The unit is configured to control the sample receiving unit to rotate about the at least one rotating shaft such that during the chemical reaction, the reaction mixture in the reaction vessel is sequentially heated separately from the at least two thermostatic modules exchange.
  • the angle between the at least one shaft and the axis of the reaction vessel is from about 80° to about 100°.
  • the angle between the at least one shaft and the axis of the reaction vessel is from about 0° to about 20°.
  • the at least two thermostat modules include a first thermostat module and a second thermostat module.
  • the reaction mixture in the reaction vessel is in heat exchange with the first thermostat module when the sample receiving unit is in the first position; and when the sample receiving unit is rotated to the second position about the at least one rotating shaft The reaction mixture in the reaction vessel is heat exchanged with the second thermostat module.
  • the temperature difference between the first thermostat module and the second thermostat module is at least about 10 degrees Celsius.
  • the first thermostat module is maintained at about 90 degrees Celsius to about 110 degrees Celsius during operation
  • the second thermostat module is maintained at about 35 degrees Celsius to about 75 degrees Celsius during operation.
  • the sample receiving unit includes at least three thermostat modules including a first thermostat module, a second thermostat module, and a third thermostat module.
  • the reaction mixture in the reaction vessel is in heat exchange with the first thermostat module when the sample receiving unit is in the first position; when the sample receiving unit is rotated to the second position about the at least one rotating shaft, The reaction mixture in the reaction vessel is in heat exchange with the second thermostat module; and when the sample receiving unit is rotated about the at least one rotating shaft to a third position, the reaction mixture in the reaction vessel is The third thermostat module performs heat exchange.
  • a temperature difference between the first thermostat module and the third thermostat module is at least about 10 degrees Celsius, and a temperature of the second thermostat module is located at Between the first thermostat module and the third thermostat module.
  • the first thermostat module is maintained at about 80 degrees Celsius to about 110 degrees Celsius during operation, and the second thermostat module is maintained at about 35 degrees Celsius during operation. About 60 degrees Celsius, and the third thermostat module is maintained at about 65 degrees Celsius to about 75 degrees Celsius during operation.
  • the chemical reaction is a nucleic acid amplification reaction.
  • the reaction vessel is a polymerase chain reaction tube.
  • each time the reaction mixture continues to exchange heat with the first thermostat module for no more than about 60 seconds the reaction mixture continues each time with the second thermostat The module performs heat exchange for no more than about 60 seconds, and the reaction mixture continues to exchange heat with the third thermostat module for no more than about 300 seconds.
  • At least a portion of the reaction mixture in the reaction vessel is not in heat exchange with any other thermostat module when it is in heat exchange with any of the at least two thermostat modules.
  • the at least two thermostatic modules are thermally insulated from one another.
  • the device of the present application further includes a reaction signal detection unit that indicates the presence and/or amount of a reaction product of the chemical reaction.
  • the reaction signal detecting unit may detect an optical signal, a spectral signal, an electrostatic signal, and/or an electrochemical signal.
  • the reaction signal detecting unit detects an optical signal, and the optical signal is a fluorescent signal.
  • the apparatus of the present application further includes an input unit in communication with the control unit, the input unit receiving an instruction from a user to perform the chemical reaction.
  • the apparatus of the present application further includes an output unit in communication with the reaction signal detecting unit, the output unit detecting the presence of the reaction product with the reaction signal detecting unit / or content information is output to the recipient.
  • the apparatus of the present application further includes a temperature sensor thermally coupled to the reaction mixture, the temperature sensor being in communication with the control unit and providing the control unit with respect to the reaction mixture Temperature information.
  • control unit is operatively coupled to a motor and the at least one rotating shaft is the motor output shaft.
  • the present application is directed to a method for conducting a chemical reaction that requires thermal cycling in at least two temperature intervals.
  • the method comprises: a) placing a reaction vessel containing a reaction mixture in a sample receiving unit rotatable about at least one axis of rotation, wherein the sample receiving unit is matched and thermally coupled to a reaction vessel for performing the chemical reaction
  • the sample receiving unit includes at least two thermostat modules, and the at least two thermostat modules are maintained in different temperature zones from each other; b) rotating the sample receiving unit about the at least one rotating shaft, thereby causing During the course of the chemical reaction, the reaction mixture in the reaction vessel is separately heat exchanged with the at least two thermostatic modules.
  • the angle between the at least one shaft and the axis of the reaction vessel is from about 70° to about 100°.
  • the angle between the at least one shaft and the axis of the reaction vessel is from about 0° to about 20°.
  • the at least two thermostat modules comprise a first thermostat module and a second thermostat module
  • the step b) comprises: b1) placing the sample receiving unit in a first position Thereby causing the reaction mixture in the reaction vessel to exchange heat with the first thermostat module; and b2) surrounding the sample receiving unit from the first location around the at least A shaft is rotated to the second position to cause heat exchange between the reaction mixture in the reaction vessel and the second thermostat module.
  • steps b1) and b2) are repeated in sequence until the chemical reaction is completed.
  • the temperature difference between the first thermostat module and the second thermostat module is at least about 10 degrees Celsius.
  • the first thermostat module is maintained at about 90 degrees Celsius to about 110 degrees Celsius during operation, and the second thermostat module is maintained at about 35 degrees Celsius to about 75 degrees Celsius.
  • the sample receiving unit includes at least three thermostat modules, the at least three thermostat modules including a first thermostat module, a second thermostat module, and a third thermostat module
  • Step b) comprises: b1) placing the sample receiving unit in a first position such that a reaction mixture in the reaction vessel is in heat exchange with the first thermostat module; b2) causing the sample receiving unit from the first a position rotated about the at least one rotating shaft to a second position such that a reaction mixture in the reaction vessel exchanges heat with the second thermostat module; and b3) causing the sample receiving unit to surround the second position
  • the at least one shaft is rotated to a third position such that the reaction mixture in the reaction vessel exchanges heat with the third thermostat module.
  • the steps b1), b2) and b3) are repeated in sequence until the chemical reaction is completed.
  • a temperature difference between the first thermostat module and the third thermostat module is at least about 10 degrees Celsius, and a temperature of the second thermostat module is located at the first thermostat module Between the third thermostat module.
  • the first thermostat module is maintained at about 90 degrees Celsius to about 110 degrees Celsius during operation, and the second thermostat module is maintained at about 35 degrees Celsius to about 60 during operation. Celsius, and the third thermostat module is maintained at about 65 degrees Celsius to about 75 degrees Celsius during operation.
  • the chemical reaction is a nucleic acid amplification reaction.
  • the reaction vessel is a polymerase chain reaction tube.
  • the reaction mixture is continuously heat exchanged with the first thermostat module for a period of no more than about 60 seconds, and the reaction mixture is continued with the second each time.
  • the thermostat module does not exchange heat for more than about 120 seconds.
  • the reaction mixture is continuously heat exchanged with the first thermostat module for no more than about 60 seconds each time, the reaction mixture continuing to the second thermostat each time.
  • the module performs heat exchange for no more than about 60 seconds, and the reaction mixture continues to exchange heat with the third thermostat module for no more than about 300 seconds.
  • At least a portion of the reaction mixture in the reaction vessel is not in heat exchange with any other thermostat module when heat exchanged with any of the at least two thermostat modules.
  • the at least two thermostatic modules are thermally insulated from one another.
  • detecting the presence and/or amount of a reaction product in the chemical reaction In certain embodiments of the methods of the present application, detecting the presence and/or amount of a reaction product in the chemical reaction.
  • the presence and/or amount of the reaction product is shown as a reaction signal, and the reaction signal includes an optical signal, a spectral signal, an electrostatic signal, and/or an electrochemical signal.
  • the reaction signal is an optical signal and the optical signal is a fluorescent signal.
  • step b) further comprises detecting the temperature of the reaction mixture and adjusting the rotation of the sample receiving unit based on the detected temperature.
  • the present application provides a system for performing a chemical reaction that requires thermal cycling in at least two temperature intervals.
  • the system can include an input module that receives a user request to perform the chemical reaction, and a reaction module responsive to the user request: 1) receiving a reaction mixture in a sample receiving unit rotatable about at least one axis of rotation a reaction vessel, wherein the sample receiving unit is matched and thermally coupled to a reaction vessel for performing the chemical reaction, the sample receiving unit includes at least two thermostat modules, and the at least two thermostat modules are maintained at a temperature zone different from each other; and 2) rotating the sample receiving unit about the at least one rotating shaft such that during the course of the chemical reaction, the reaction mixture in the reaction vessel is sequentially and the at least two
  • the thermostat module separately performs heat exchange; and an output module operatively coupled to the reaction module, wherein the output module outputs information regarding the presence and/or content of the reaction product in the chemical reaction to the recipient.
  • the present application provides a system for performing a chemical reaction that requires thermal cycling in at least two temperature intervals.
  • the system can include an input module that receives a user request to perform the chemical reaction, a reaction module that performs the method described herein in response to the user request, and an output module operatively coupled to the A reaction module, wherein the output module outputs information about the presence and/or content of a reaction product in the chemical reaction to a recipient.
  • the present application provides a computer readable medium containing machine executable code, when executed by one or more computer processors, to perform a method of performing a chemical reaction, the method comprising: A reaction vessel containing the reaction mixture is placed in a sample receiving unit rotatable about at least one axis of rotation, wherein the sample receiving unit is mated and thermally coupled to a reaction vessel for performing the chemical reaction, the sample receiving unit comprising At least two thermostat modules, and the at least two thermostat modules are maintained in different temperature zones from each other; The sample receiving unit is rotated about the at least one rotating shaft such that the reaction mixture in the reaction vessel is sequentially heat exchanged with the at least two thermostatic modules, respectively, during the course of the chemical reaction.
  • the present application provides a computer readable medium containing machine executable code that, when executed by one or more computer processors, implements the methods described herein.
  • the present application provides a system for performing a chemical reaction, comprising: an electronic display screen including a user interface displaying graphical elements that are accessible by a user to perform a chemical reaction And a computer processor operatively coupled to the electronic display screen and programmed to perform the chemical reaction when the user selects the graphical element, the chemical reaction scheme comprising: enabling A reaction vessel containing the reaction mixture is placed in a sample receiving unit rotatable about at least one axis of rotation, wherein the sample receiving unit is mated and thermally coupled to a reaction vessel for performing the chemical reaction, the sample receiving unit comprising At least two thermostatic modules, and the at least two thermostatic modules are maintained in different temperature zones from each other; and rotating the sample receiving unit about the at least one rotating shaft such that during the course of the chemical reaction, The reaction mixture in the reaction vessel is in turn heat exchanged with the at least two thermostatic modules, respectively.
  • the present application provides a system for performing a chemical reaction, comprising: an electronic display screen including a user interface displaying graphical elements that are accessible by a user to perform a method for performing a chemical reaction ;with
  • a computer processor operatively coupled to the electronic display screen and programmed to perform the methods described herein when the user selects the graphical element.
  • FIG. 1A, 1B, 1C show cross-sectional views of one embodiment of the apparatus of the present application at different angles of rotation.
  • Figure 2 shows a perspective view of one embodiment of the apparatus of the present application.
  • Figure 3 shows a perspective view of one embodiment of the apparatus of the present application.
  • Figure 4 shows a cross-sectional view of one embodiment of the apparatus of the present application.
  • Figures 5A, 5B, and 5C show cross-sectional views of one embodiment of the apparatus of the present application at different angles of rotation.
  • Figure 6 shows a perspective view of one embodiment of the apparatus of the present application.
  • FIG. 7A, 7B show cross-sectional views of one embodiment of the apparatus of the present application.
  • nucleic acid amplification generally refers to the replication of one or more nucleic acid sequences to form one or more "copy” or “amplification products" of a nucleic acid.
  • DNA amplification generally refers to the replication of one or more DNA sequences to form one or more "copy” or “amplified DNA products" of a DNA molecule.
  • reverse transcription amplification generally refers to the action of reverse transcriptase from the ribonucleoside
  • the acid (RNA) template forms deoxyribonucleic acid (DNA).
  • cycle threshold generally refers to a cycle in a thermal cycle in which the increase in detectable signal produced by the amplification product reaches a statistically significant higher than the background. The level of the signal.
  • denaturation generally refers to the complete or partial unwinding of the helical structure of a double-stranded nucleic acid, and in some cases refers to the unwinding of the secondary structure of a single-stranded nucleic acid. In some cases, denaturation may also include inactivation of the pathogen cell wall or viral envelope, as well as inactivation of the protein inhibitor. Conditions that affect denaturation can include “denaturation temperature” and “denaturation duration.” “Denaturing temperature” generally refers to the temperature at which denaturation is allowed to occur, and “denaturation duration” generally refers to the length of time consumed for denaturation or the length of time at which the denaturation temperature is maintained.
  • extension generally refers to the incorporation of nucleotides into a nucleic acid in a template-directed manner. Extension can occur by means of an enzyme such as a polymerase or a reverse transcriptase. Conditions that affect the extension may include “extension temperature” and “extension duration”, “extension temperature” generally refers to the temperature at which elongation is allowed to occur, and “extension duration” generally refers to the length of time it takes for extension to occur.
  • annealing and “hybridization” are used interchangeably and generally refer to the nucleotide base-pairing interaction of one nucleic acid with another nucleic acid, resulting in a double-stranded structure, a triple-stranded structure. Or other more advanced structures.
  • Conditions affecting annealing may include “annealing temperature” and “annealing duration”.
  • Annealing temperature generally refers to the temperature at which annealing is allowed to occur
  • annealing duration generally refers to the length of time it takes for annealing to occur.
  • nucleic acid generally refers to a polymeric form of nucleotides of any length (deoxyribonucleotides (dNTPs) or ribonucleotides (rNTPs)) or analogs thereof.
  • the nucleic acid can have any three-dimensional structure and can perform any known or unknown function.
  • nucleic acids include coding regions or non-coding regions of DNA, RNA, genes or gene fragments, one or more loci, exons, introns, determined by linkage analysis, Messenger RNA (mRNA), transport RNA, ribosomal RNA, short interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA), ribozyme, cDNA, recombinant nucleic acid, Branched nucleic acids, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes and primers.
  • mRNA Messenger RNA
  • transport RNA transport RNA
  • ribosomal RNA short interfering RNA
  • shRNA short hairpin RNA
  • miRNA microRNA
  • ribozyme ribozyme
  • cDNA recombinant nucleic acid
  • Branched nucleic acids plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence
  • a nucleic acid can comprise one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs.
  • the nucleotide sequence of the nucleic acid can be interrupted by a non-nucleotide component.
  • the nucleic acid can be further modified (eg, by coupling or binding to a reporter) after polymerization.
  • primer extension reaction generally refers to the process of double-stranded nucleic acid denaturation, binding of a primer to one or both strands of a denatured nucleic acid (also “annealing” or “hybridization”), followed by primer extension.
  • reaction mixture generally refers to a composition comprising reagents necessary for completion of nucleic acid amplification (eg, DNA amplification, RNA amplification), non-limiting examples of such reagents include for target RNA or Target DNA has a specific primer set, DNA produced by reverse transcription of RNA, DNA polymerase, reverse transcriptase (for example, reverse transcription for RNA), suitable buffer (including zwitterionic buffer), cofactor (eg, divalent and monovalent cations), dNTPs, and other enzymes (eg, uracil-DNA glycosylase (UNG), etc.).
  • the reaction mixture may also contain one or more reporters.
  • the reaction mixture can include an emulsion or a packaged composition.
  • reporter generally refers to a composition that produces a detectable signal, the presence or absence of which can be used to indicate the presence or absence of an amplification product.
  • target nucleic acid generally refers to a nucleic acid molecule having a certain nucleotide sequence in the starting population of a nucleic acid molecule, which needs to be determined for one or more of its presence, amount and/or sequence. The change.
  • the target nucleic acid can be any type of nucleic acid, including DNA, RNA, and analogs thereof.
  • target ribonucleic acid (RNA) is generally referred to as a target nucleic acid of RNA.
  • target deoxyribonucleic acid (DNA) is generally referred to as a target nucleic acid of DNA.
  • the term "subject” generally refers to an entity having genetic information that is testable or detectable.
  • the subject can be a human.
  • the subject can be a vertebrate, such as a mammal.
  • Non-limiting examples of mammals include rats, baboons, monkeys, pigs, cows, sheep, and the like.
  • Subjects can also be, for example, food, plants, soil, and water.
  • thermal cycle generally refers to the process of repetitively changing a reaction system (eg, a mixture of chemical reactions) between two or more different temperatures.
  • axis and “rotational axis” are used interchangeably and generally refer to a geometric line around which an object rotates. In some embodiments, the vertical distance between any point in the object and the axis of rotation remains constant as the object is rotated about the axis of rotation.
  • sample receiving unit generally refers to an assembly of a reaction vessel capable of carrying and/or immobilizing a sample or containing a sample.
  • matching generally refers to the need for the shape, size and or material of an object or object or substance to interact with one another. In some cases, “matching” may also mean the degree of similarity or complementarity between nucleic acid sequences.
  • thermally coupled generally refers to the association of two or more physical systems and/or components in a particular manner to transfer thermal energy and heat between the systems and/or components.
  • thermometer module generally refers to a module that maintains a substantially constant temperature during the course of the reaction or operation of the device.
  • heat exchange generally refers to the process of thermal energy transfer between two objects or portions of the same object due to temperature differences.
  • heat exchange can be performed by heat conduction, heat convection, and/or heat radiation.
  • axis generally refers to a line that divides a planar or three-dimensional structure into symmetrical portions.
  • the "axis” generally refers to the longest straight line that divides the three-dimensional structure into symmetrical portions.
  • nucleic acid amplification reaction generally refers to a chemical reaction that causes amplification of a nucleic acid.
  • the nucleic acid amplification reaction can be a polymerase chain reaction.
  • polymerase chain reaction tube generally refers to a tube suitable for causing a reaction mixture to undergo a polymerase chain reaction therein.
  • the polymerase chain reaction tube may be a microcentrifuge tube or a PCR tube.
  • thermal insulation generally means reducing or preventing two due to an increase in thermal resistance. Thermal energy transfer between more or more objects or parts of the same object.
  • reaction signal detection unit generally refers to a device or component for detecting or indicating the presence and/or intensity of a reaction signal.
  • informational communication generally refers to the ability of information to flow and/or communicate between two or more objects or between portions of the same object.
  • temperature sensor generally refers to a device that is capable of sensing temperature or temperature changes and converting it into an effective output signal.
  • control unit generally refers to a component that is capable of controlling one or more parameters of a device or system.
  • output shaft generally refers to a shaft for outputting power
  • motor output shaft may be a shaft that is coupled to a motor and that is used to output motor power.
  • operably connected generally refers to the manner in which two objects or portions of an object are connected such that an object or portion can achieve a predetermined effect on another object or portion.
  • the term "about” generally means a range of 0.5% to 10% above or below a specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5% above or below a specified value, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% range.
  • the present application provides a device for performing a chemical reaction that requires thermal cycling between at least two temperature zones (eg, two target temperatures).
  • the apparatus can include a sample receiving unit rotatable about at least one axis of rotation, wherein the sample receiving unit is mated and thermally coupled to a reaction vessel for performing the chemical reaction.
  • the sample receiving unit may include at least two thermostat modules, and the at least two thermostat modules are maintained in different temperature zones from each other.
  • the sample receiving unit comprises two, three, four or more thermostat modules.
  • the apparatus may further include a control unit configured to control the sample connection
  • the receiving unit is rotated about the at least one rotating shaft such that during the progress of the chemical reaction, the reaction mixture in the reaction vessel is sequentially heat exchanged with the at least two thermostatic modules, respectively.
  • the device of the present application can include a thermal cycler for performing a nucleic acid amplification reaction (eg, a PCR reaction).
  • a thermal cycler for performing a nucleic acid amplification reaction (eg, a PCR reaction).
  • 1A, 1B, 1C show cross-sectional views of an exemplary device of the present application.
  • the apparatus includes a sample receiving unit 100 that includes a first thermostat module 104 and a second thermostat module 105 and a thermal insulation component 102.
  • the thermal insulation component 102 thermally isolates the first thermostat module 104 from the second thermostat module 105 and between the sample receiving unit 100 and the external environment.
  • the sample receiving unit 100 is matched and thermally coupled to a reaction vessel (e.g., PCR tube) 101 containing a reaction mixture 103.
  • a reaction vessel e.g., PCR tube
  • the reaction mixture 103 exchanges heat with the first thermostat module 104 for the required time. Subsequently, when the sample receiving unit 100 rotates about the rotating shaft and passes through the intermediate position (as shown in FIG. 1B) to the second position (as shown in FIG. 1C), the reaction mixture 103 exchanges heat with the second thermostat module 105 and continues to be required. time.
  • the sample receiving unit 100 may reciprocally rotate between the first position and the second position as needed until a desired reaction product (for example, a nucleic acid amplification product) is obtained.
  • FIG. 2 shows a perspective view of an exemplary device 200 of the present application.
  • the apparatus 200 includes a sample receiving unit 209 rotatable about at least one axis of rotation 203 that matches a reaction vessel (eg, PCR tube) 201 for performing a chemical reaction (eg, a nucleic acid amplification reaction) and is hot coupling.
  • the sample receiving unit 209 includes a first thermostat module 204 and a second thermostat module 205 and a thermal insulation component 202.
  • the thermal insulation component 202 thermally isolates the first thermostat module 204 from the second thermostat module 205 and between the sample receiving unit 209 and the external environment.
  • the apparatus 200 further includes a control unit (eg, a motor) 207 that controls rotation of the sample receiving unit 209 about the at least one rotating shaft 203, and between the axis of the reaction vessel 201 and the rotating shaft 203 during rotation The angle is about 90°.
  • the control unit 207 is fixed to the base 208 by a fixing member 206.
  • FIG. 3 shows a perspective view of an exemplary device 300 of the present application.
  • the apparatus 300 includes a sample receiving unit 309 rotatable about at least one axis of rotation 305 that matches a reaction vessel (eg, PCR tube) 301 for performing a chemical reaction (eg, a nucleic acid amplification reaction) and is hot coupling.
  • the sample receiving unit 309 includes a first thermostat module 302, a second thermostat module 303, and a third thermostat module 304.
  • the apparatus 300 also includes a control unit (eg, a motor) 307 that controls rotation of the sample receiving unit 309 about the at least one rotating shaft 305.
  • the control unit 307 is fixed to the base 308 by a fixing member 306.
  • FIG. 4 shows a cross-sectional view of an exemplary device 400 of the present application.
  • the apparatus 400 includes a sample receiving unit 409 rotatable about at least one axis of rotation 405 that matches a reaction vessel (eg, PCR tube) 401 for performing a chemical reaction (eg, a nucleic acid amplification reaction) and is hot Coupling, the reaction vessel 402 is contained in the reaction vessel 401.
  • the sample receiving unit 409 includes thermostat modules 403 and 404.
  • the apparatus 400 also includes a control unit (eg, a motor) 407 that controls rotation of the sample receiving unit 409 about the at least one shaft 405.
  • the control unit 407 is fixed to the base 408 by a fixing member 406. As shown in FIG.
  • the angle between the axis of the reaction vessel 401 and the rotating shaft 405 is about 0.
  • heat exchange occurs between the reaction mixture 402 and the thermostat module 404 for the required time.
  • the apparatus includes a sample receiving unit 500 that includes a first thermostat module 504, a second thermostat module 505, a third thermostat module 506, and a thermal insulation component 502.
  • the thermal insulation component 502 thermally isolates the first thermostat module 504, the second thermostat module 505, and the third thermostat module 506 and between the sample receiving unit 500 and the external environment.
  • the sample receiving unit 500 is matched and thermally coupled to a reaction vessel (eg, PCR tube) 501 containing a reaction mixture 503.
  • a reaction vessel eg, PCR tube
  • the sample receiving unit 500 is rotated about the rotation axis to the second position (as shown in FIG. 5B), at which time the reaction mixture 503 is heat exchanged with the second thermostat module 505 for the required time. Then, the sample receiving unit 500 continues to rotate about the rotation axis to the third position (as shown in FIG. 5C), at which time the reaction mixture 503 exchanges heat with the first thermostat module 504 for the required time.
  • the sample receiving unit 500 may reciprocally rotate between the first position, the second position, and the third position as needed until a desired reaction product (for example, a nucleic acid amplification product) is obtained.
  • FIG. 6 shows a perspective view of an exemplary device 600 of the present application.
  • the apparatus includes a sample receiving unit 609 that includes a first thermostat module 604, a second thermostat module 605, a third thermostat module 606, and a thermal insulation component 602.
  • the thermal insulation component 602 thermally isolates the first thermostat module 604, the second thermostat module 605, and the third thermostat module 606 and between the sample receiving unit 609 and the external environment.
  • the apparatus 600 also includes a control unit (eg, a motor) 603 that controls rotation of the sample receiving unit 609 about the at least one shaft 601.
  • the control unit 603 is fixed to the base 608 by a fixing member 607.
  • the apparatus includes a sample receiving unit 707 that includes a first thermostat module 703 and a second thermostat module 704.
  • the sample receiving unit 707 is matched and thermally coupled to a reaction vessel (eg, PCR tube) 701 containing a reaction mixture 702.
  • the apparatus 700 further includes an excitation component 706 and a signal detection component 705 that can emit excitation light such that the reporter in the reaction mixture 702 produces a detectable signal (eg, a fluorescent signal) for signal detection component 705. Detection.
  • the present application provides a method for performing a chemical reaction that requires thermal cycling between at least two temperature zones (eg, two target temperatures).
  • the method can include placing a reaction vessel containing the reaction mixture in a rotatable manner about at least one axis of rotation a sample receiving unit, wherein the sample receiving unit is matched and thermally coupled to a reaction vessel for performing the chemical reaction, the sample receiving unit includes at least two thermostat modules, and the at least two thermostat modules are maintained In different temperature zones from each other.
  • the method may further include rotating the sample receiving unit about the at least one rotating shaft such that during the progress of the chemical reaction, the reaction mixture in the reaction vessel is sequentially separated from the at least two thermostatic modules Perform heat exchange.
  • the methods of the present application can be used to perform nucleic acid amplification reactions, such as polymerase chain reaction (PCR), which require thermal cycling in at least two temperature intervals.
  • a reaction vessel eg, a PCR tube
  • the sample receiving unit comprising at least two thermostat modules and the at least two thermostat modules being maintained In different temperature zones from each other.
  • the thermal cycle can be achieved by rotating the sample receiving unit about the at least one rotating shaft such that the reaction mixture in the reaction vessel is separately heat exchanged with the at least two thermostatic modules, thereby heating Or cooling the reaction mixture to a target temperature (eg, a predetermined reaction temperature).
  • a target temperature eg, a predetermined reaction temperature
  • the PCR reaction can involve a set reaction temperature of 94 ° C and 64 ° C
  • the sample receiving unit can include a first thermostat module maintained at about 94 ° C and a second thermostat module maintained at about 64 ° C.
  • the sample receiving unit can be rotated about the at least one rotating shaft to cause heat exchange between the reaction mixture in the reaction vessel and the first thermostat module, and the temperature of the reaction mixture rapidly reaches about 94 ° C.
  • the sample receiving unit can be rotated again around the at least one rotating shaft to cause heat exchange between the reaction mixture in the reaction vessel and the second thermostat module, and the temperature of the reaction mixture is rapidly lowered to about 64 °C.
  • the present application provides a system for performing a chemical reaction that requires thermal cycling between at least two temperature zones (eg, two target temperatures).
  • the system can include an input module that receives a user request to perform the chemical reaction.
  • the system can also include a reaction module that can be wrapped around in response to the user request Receiving a reaction vessel containing the reaction mixture in a sample receiving unit that rotates less than one of the shafts, wherein the sample receiving unit is matched and thermally coupled to a reaction vessel for performing the chemical reaction, the sample receiving unit including at least two thermostats Modules, and the at least two thermostat modules are maintained in different temperature zones from each other.
  • the reaction module may also rotate the sample receiving unit about the at least one rotating shaft in response to the user request, such that during the progress of the chemical reaction, the reaction mixture in the reaction vessel is sequentially
  • the at least two thermostat modules are respectively heat exchanged.
  • the system can also include an output module operatively coupled to the reaction module, wherein the output module outputs information regarding the presence and/or amount of reaction products in the chemical reaction to a recipient.
  • the present application provides a computer readable medium containing machine executable code that, when executed by one or more computer processors, implements a method of performing a chemical reaction.
  • the method includes placing a reaction vessel containing a reaction mixture in a sample receiving unit rotatable about at least one axis of rotation, wherein the sample receiving unit is matched and thermally coupled to a reaction vessel for performing the chemical reaction,
  • the sample receiving unit includes at least two thermostat modules, and the at least two thermostat modules are maintained in different temperature zones from each other.
  • the method may further include rotating the sample receiving unit about the at least one rotating shaft such that during the progress of the chemical reaction, the reaction mixture in the reaction vessel is sequentially separated from the at least two thermostatic modules Perform heat exchange.
  • the present application provides a system for performing a chemical reaction
  • the system can include an electronic display screen including a user interface displaying graphical elements that are accessible by a user for performing chemistry Reaction scheme.
  • the system can also include a computer processor operatively coupled to the electronic display screen and programmed to perform the chemical reaction scheme when the user selects the graphical element.
  • the chemical reaction scheme may include placing a reaction vessel containing the reaction mixture in a sample receiving unit rotatable about at least one rotating shaft, wherein the sample receiving unit matches a reaction vessel for performing the chemical reaction and Thermally coupled, the sample receiving unit includes at least two thermostat modules, and the at least two constant The temperature modules are maintained in different temperature zones from each other.
  • the chemical reaction scheme may further include rotating the sample receiving unit about the at least one rotating shaft such that during the progress of the chemical reaction, the reaction mixture in the reaction vessel is sequentially and the at least two The thermostat modules are separately exchanged for heat.
  • the angle between the at least one shaft and the axis of the reaction vessel may be from about 0° to about 360°, such as from about 0° to about 5°, about 0°. Up to about 10°, from about 0° to about 20°, from about 0° to about 30°, from about 0° to about 40°, from about 0° to about 50°, from about 0° to about 60°, from about 0° to about 70°, from about 0° to about 80°, from about 0° to about 90°, from about 0° to about 100°, from about 0° to about 110°, from about 0° to about 120°, from about 0° to about 130° , from about 0° to about 140°, from about 0° to about 150°, from about 0° to about 160°, from about 0° to about 170°, from about 0° to about 180°, from about 0° to about 190°, about 0° to about 200°
  • the angle between the at least one shaft and the axis of the reaction vessel is from about 70° to about 110°, such as from about 80° to about 100°, from about 85° to about 95°, From about 85° to about 90°, or about 90°. In certain embodiments, the angle between the at least one shaft and the axis of the reaction vessel is from about 0 to 30, such as from about 0 to about 25, from about 0 to about 20, about 0° to about 15°, from about 0° to about 10°, from about 0° to about 5°, or about 0°.
  • the at least two thermostat modules comprise a first thermostat module and a second thermostat module, and wherein the first thermostat module and the second thermostat module are The devices, methods, or systems are maintained in different temperature zones when they are in operation or during operation.
  • the reaction mixture in the reaction vessel can be heat exchanged with the first thermostat module.
  • the reaction mixture in the reaction vessel may be The second thermostat module performs heat exchange.
  • the chemical reaction can be a nucleic acid amplification reaction.
  • the first thermostat module can be maintained at a denaturation temperature of the nucleic acid amplification reaction while the apparatus, method or system is operating or performing, and the second thermostat module can be maintained at an extension temperature of the nucleic acid amplification reaction,
  • the denaturation temperature can be different from the extension temperature.
  • the denaturation temperature can range from about 80 ° C to about 110 ° C, from about 90 ° C to about 110 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 97 ° C, such as from about 92 ° C to about 95 ° C.
  • the denaturation temperature is at least about 80 ° C, at least about 81 ° C, at least about 82 ° C, at least about 83 ° C, at least about 84 ° C, at least about 85 ° C, at least about 86 ° C, at least about 87 ° C, at least about 88 ° C, at least about 89 ° C, at least about 90 ° C, at least about 91 ° C, at least about 92 ° C, at least about 93 ° C, at least about 94 ° C, at least about 95 ° C, at least about 96 ° C, at least about 97 °C, at least about 98 ° C, at least about 99 ° C or at least about 100 ° C.
  • the extension temperature can range from about 30 °C to about 80 °C. In certain embodiments, the extension temperature is from about 35 °C to about 75 °C. For example, the extension temperature can range from about 35 ° C to about 72 ° C, from about 40 ° C to about 65 ° C, from about 45 ° C to about 60 ° C, from about 50 ° C to about 60 ° C.
  • the extension temperature is at least about 35 ° C, at least about 36 ° C, at least about 37 ° C, at least about 38 ° C, at least about 39 ° C, at least about 40 ° C, at least about 41 ° C, at least about 42 ° C, At least about 43 ° C, at least about 44 ° C, at least about 45 ° C, at least about 46 ° C, at least about 47 ° C, at least about 48 ° C, at least about 49 ° C, at least about 50 ° C, at least about 51 ° C, at least about 52 ° C, At least about 53 ° C, at least about 54 ° C, at least about 55 ° C, at least about 56 ° C, at least about 57 ° C, at least about 58 ° C, at least about 59 ° C, at least about 60 ° C, at least about 61 ° C, at least about 62 ° C, At least about 63 ° C, at least about 64 ° C,
  • the sample receiving unit may include at least three thermostat modules including a first thermostat module, a second thermostat module, and a third thermostat module, And the first thermostat module, the second thermostat module, and the third thermostat module are maintained in different temperature zones from each other while the device, method, or system is operating or in progress.
  • the reaction mixture in the reaction vessel can be heat exchanged with the first thermostat module.
  • the reaction mixture in the reaction vessel may be in heat exchange with the second thermostat module when the sample receiving unit is rotated about the at least one rotating shaft to the second position.
  • the reaction mixture in the reaction vessel is in heat exchange with the third thermostat module when the sample receiving unit is rotated about the at least one rotating shaft to a third position.
  • the chemical reaction can be a nucleic acid amplification reaction.
  • the first thermostat module can be maintained at a denaturation temperature of the nucleic acid amplification reaction while the device is in operation, the second thermostat module can be maintained at an annealing temperature and an extension temperature of the nucleic acid amplification reaction, the third The thermostat module can be maintained at an extension temperature of the nucleic acid amplification reaction.
  • the denaturation temperature can be from about 80 ° C to about 110 ° C, from about 90 ° C to about 110 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 97 ° C, such as from about 92 ° C to about 95 ° C, at least About 80 ° C, at least about 81 ° C, at least about 82 ° C, at least about 83 ° C, at least about 84 ° C, at least about 85 ° C, at least about 86 ° C, at least about 87 ° C, at least about 88 ° C, at least about 89 ° C, at least About 90 ° C, at least about 91 ° C, at least about 92 ° C, at least about 93 ° C, at least about 94 ° C, at least about 95 ° C, at least about 96 ° C, at least about 97 ° C, at least about 98 ° C, at least about 99 ° C or at
  • the annealing temperature can range from about 30 °C to about 65 °C. In certain embodiments, the annealing temperature is from about 30 °C to about 60 °C. For example, the annealing temperature can range from about 35 ° C to about 60 ° C, from about 40 ° C to about 60 ° C, from about 45 ° C to about 60 ° C, from about 50 ° C to about 60 ° C. In certain embodiments, the annealing temperature can be at least about 45 ° C, at least about 46 ° C, at least about 47 ° C, at least about 48 ° C, at least about 49 ° C, at least about 50 ° C, at least about 51 ° C, at least about 52 ° C.
  • the extension temperature can range from about 60 °C to about 80 °C. In certain embodiments, the extension temperature is from about 65 °C to about 80 °C. For example, the extension temperature can range from about 65 ° C to about 75 ° C, from about 70 ° C to about 80 ° C, about 70 ° C to about 75 ° C.
  • the extension temperature can be at least about 65 ° C, at least about 66 ° C, at least about 67 ° C, at least about 68 ° C, at least about 69 ° C, at least about 70 ° C, at least about 71 ° C, at least about 72 ° C. At least about 73 ° C, at least about 74 ° C, at least about 75 ° C, at least about 76 ° C, at least about 77 ° C, at least about 78 ° C, at least about 79 ° C, or at least about 80 ° C.
  • the reaction vessel may be a polymerase chain reaction tube.
  • the time during which the reaction mixture continues to exchange heat with the first thermostat module may be no more than about 5 seconds, no more than about 10 seconds, no more than about 15 seconds, Not more than about 20 seconds, no more than about 25 seconds, no more than about 30 seconds, no more than about 35 seconds, no more than about 40 seconds, no more than about 45 seconds, no more than about 50 seconds, no more than about 55 seconds, no more than About 60 seconds, no more than about 70 seconds, no more than about 80 seconds, or no more than about 90 seconds.
  • the reaction mixture may be in heat exchange with the second thermostat module for a period of no more than about 5 seconds, no more than about 10 seconds, no more than about 15 seconds, no more than about 20 Seconds, no more than about 25 seconds, no more than about 30 seconds, no more than about 35 seconds, no more than about 40 seconds, no more than about 45 seconds, no more than about 50 seconds, no more than about 60 seconds, no more than about 70 seconds, No more than about 80 seconds, no more than about 90 seconds, no more than about 100 seconds, no more than about 110 seconds, no more than about 120 seconds, no more than about 130 seconds, no more than about 140 seconds, or no more than about 150 seconds.
  • the reaction mixture may be in heat exchange with the third thermostat module for a period of no more than about 5 seconds, no more than about 10 seconds, no more than about 15 seconds, no more than about 20 Seconds, no more than about 25 seconds, no more than about 30 seconds, no more than about 35 seconds, no more than about 40 seconds, no more than about 45 seconds, no more than about 50 seconds, no more than about 60 seconds, no more than about 70 seconds, No more than about 80 seconds, no more than about 90 seconds, no more than about 100 seconds, no more than about 110 seconds, no more than about 120 seconds, no more than about 130 seconds, no more than about 140 seconds, or no more than about 150 seconds.
  • At least a portion of the reaction mixture in the reaction vessel is substantially not in heat exchange with any other thermostat module when it is in heat exchange with any of the at least two thermostat modules.
  • a reaction signal detecting unit or a detecting step indicating the presence and/or content of the reaction product of the chemical reaction may also be included.
  • the reaction signal detecting unit or detecting step can detect an optical signal, a spectral signal, an electrostatic signal, and/or an electrochemical signal.
  • the reaction signal detection unit or detection step detects an optical signal, such as a fluorescent signal.
  • an input unit or input module in communication with the control unit may also be included, the input unit or input module being operable to receive an instruction from the user regarding the chemical reaction.
  • an output unit or an output module that is in communication with the reaction signal detecting unit may be further included, and the output unit or the output module may detect the reaction detected by the reaction signal detecting unit Information on the presence and/or content of the product is output to the recipient.
  • a temperature sensor thermally coupled to the reaction mixture, the temperature sensor being communicable with the control unit and providing the control unit with temperature regarding the temperature of the reaction mixture information.
  • control unit is operably connectable to a motor.
  • the at least one rotating shaft is the motor output shaft.
  • the reaction signal can include an optical signal, a spectral signal, an electrostatic signal, and/or an electrochemical signal.
  • the reaction signal is an optical signal and the optical signal can be a fluorescent signal.
  • an instruction for performing the chemical reaction may be transmitted through the input unit.
  • the storage of the reaction product may also be included
  • the information at and/or content is output to the recipient.
  • it may further comprise detecting the temperature of the reaction mixture and adjusting the rotation of the sample receiving unit in accordance with the detected temperature.
  • the methods, devices, and systems of the present application can precisely control temperature changes in a chemical reaction system under conditions of low energy consumption.
  • the gradual change time i.e., the time it takes for the thermal cycler to switch the sample or reaction mixture from one temperature to another
  • the rate of grading can be a critical factor.
  • the time required for the amplification reaction to produce a detectable, amplified signal indicative of the presence or amount of target nucleic acid can be shortened by reducing the gradation time or accelerating the rate of grading.
  • the ramping time and/or the ramp rate can be different between different cycles.
  • the gradation time and/or the grading rate are the same between different cycles. The gradation time and/or the grading rate can be adjusted depending on the type of sample and/or the composition of the reaction mixture.
  • the gradation time and/or the gradual rate can be controlled by adjusting the time during which the reaction mixture contacts or exchanges heat with the thermostat module of the present application.
  • the gradation time and/or the grading rate can be controlled by increasing the contact area between the reaction mixture and the thermostat module of the present application.
  • the gradation time and/or the gradual rate can be controlled by setting the temperature of the thermostat module of the present application to be above and/or below the target temperature.
  • the rate of gradual change of the temperature of the reaction mixture can be at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25 or 30 ° C / sec.
  • the reaction mixture can be subjected to heat exchange with the thermostat module for at least about 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 seconds, or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35 , 40, 45, 50, 55, or 60 minutes.
  • the reaction mixture can be subjected to heat exchange with the thermostat module for a time of at most about 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 or 60 seconds, or up to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 or 60 minutes.
  • the chemical reaction may include at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more cycles .
  • the chemical reaction can include no more than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45 or 50 times.
  • the rotation of the sample receiving unit about the at least one rotating shaft can be adjusted by the control unit, which can be adjusted mechanically or electronically.
  • the control unit includes one or more electric machines.
  • rotation of the sample receiving unit about the at least one axis of rotation can be controlled by an electronic control system.
  • the electronic control system can include one or more computer processors operatively coupled to one or more motors to adjust the sample receiving unit to rotate in a particular direction and speed .
  • the rotation of the sample receiving unit about the at least one axis of rotation is controlled by a mechanical control system, for example, the sample receiving unit can be adjusted to rotate in a particular direction and speed by gears or tracks or the like.
  • each thermostat module can be independently of each other with each reaction cycle for about 0.1 second, about 0.2 seconds, about 0.3 seconds, about 0.4 seconds, about 0.5 seconds, about 0.6 seconds, about 0.7 seconds, about 0.7 seconds.
  • the time required to complete a thermal cycle can be less than or equal to about 10 minutes, about 5 minutes, about 1 minute, about 50 seconds, about 40 seconds, about 30 seconds, about 20 seconds, about 15 seconds, about 10 seconds, about 9 seconds, about 8 seconds, about 7 seconds, about 6 seconds, about 5 seconds, about 4 seconds, about 3 seconds, about 2 seconds, about 1 second, about 0.9 seconds About 0.8 seconds, about 0.7 seconds, about 0.6 seconds, about 0.5 seconds, about 0.4 seconds, about 0.3 seconds, about 0.2 seconds, or about 0.1 seconds.
  • parameters related to thermal cycling in the chemical reaction such as the temperature of the thermostatic module, the target temperature during the reaction, and the reaction mixture, may be controlled by programming or other means of adjustment.
  • the chemical reaction carried out by the apparatus, method or system of the present application can be carried out in one or more reaction mixtures.
  • the reaction mixture can comprise a target nucleic acid.
  • the reaction mixture may also comprise one or more reporters (eg, detectable nucleic acid binding reagents) that detect the amplification product.
  • the reaction mixture may further comprise one or more reagents for performing nucleic acid amplification, for example, a nucleic acid polymerase, a reverse transcriptase, dNTP or Mg 2+ ions, and the like.
  • the reaction mixture is an emulsion or emulsion.
  • the reaction mixture is encapsulated.
  • Biological samples can also be included in the reaction mixture.
  • the biological sample can be taken from a subject.
  • the biological sample is taken directly from a living subject.
  • the biological sample can include, for example, blood, urine, feces, saliva, cerebrospinal fluid, and sweat, and the like.
  • Any biological sample comprising a nucleic acid can be used, which can be in solid form (e.g., biological tissue) or in liquid form (e.g., body fluid).
  • Non-limiting examples of biological samples include blood obtained from any anatomical location of the subject (eg, tissue, circulatory system, bone marrow) (or components of blood - eg, white blood cells, red blood cells, platelets), from the subject Cell, skin, heart, lung, kidney, exhaled breath, bone marrow, feces, semen, vaginal fluid, tissue fluid derived from tumor tissue, breast, pancreas, cerebrospinal fluid, tissue, throat swab, biopsy obtained from any anatomical location Placental fluid, amniotic fluid, liver, muscle, smooth muscle, bladder, gallbladder, colon, intestine, brain, cavity fluid, sputum, pus, microbiota, meconium, milk, prostate, esophagus, thyroid, serum, saliva, Urine, gastric and digestive juices, tears, eye fluids, sweat, mucus, earwax, oil, glandular secretions, spinal fluid, hair, nails, skin cells, plasma, nasal swabs or
  • the subject can be a living individual or a dead individual.
  • the subject can be a human or an animal.
  • the subject can be a mammal, such as a donkey, a monkey, a cat, a dog, a pig, a cow, a sheep, a rat, and the like.
  • the subject can be food, soil, water, or air, and the like.
  • a biological sample taken directly from a subject generally refers to a biological sample that has not been further processed.
  • blood can be taken from the subject's peripheral circulatory system (eg, via a blood collection needle) and added directly to the reaction mixture.
  • An anticoagulant may be included in the reaction mixture such that the blood is available for further analysis.
  • a swab can be used to obtain epithelial cells from the oral epithelium or other epithelial tissue of the subject and after the biological sample from the subject is obtained, the swab containing the biological sample can be immersed A liquid (eg, a buffer) is added to collect biological samples from the swab.
  • the biological sample can be pretreated (eg, purified, nucleic acid extracted, etc.) prior to being added to the reaction mixture.
  • the methods, systems or devices of the present application can be used to amplify a target nucleic acid to generate an amplification product.
  • the target nucleic acid can be a target RNA or a target DNA.
  • the target RNA can be any type of RNA.
  • the target RNA is viral RNA.
  • viral RNA may be pathogenic to a subject.
  • pathogenic viral RNA include human immunodeficiency virus I (HIV I), human immunodeficiency virus II (HIV II), orthomyxovirus, Ebola virus, dengue virus, influenza virus (eg, H1N1) H3N2, H7N9 or H5N1), hepatitis virus, hepatitis A virus, hepatitis B virus, hepatitis C virus (eg, armored RNA-HCV virus), hepatitis D virus, hepatitis E virus, hepatitis G virus, EB Virus, mononucleosis virus, cytomegalovirus, SARS virus, West Nile fever virus, poliovirus and measles virus.
  • the target DNA can be any type of DNA.
  • the target DNA is viral DNA.
  • viral DNA may be pathogenic to a subject.
  • Non-limiting examples of DNA viruses include herpes simplex virus, variola virus, adenovirus (eg, adenovirus type 55, adenovirus type 7) and varicella virus (eg, fowl pox).
  • the target DNA can be bacterial DNA.
  • the bacterial DNA may be derived from a bacterium that is pathogenic to the subject, for example, Mycobacterium tuberculosis (a bacterium known to cause tuberculosis).
  • the target DNA can be DNA from a pathogenic protozoan (eg, one or more protozoa of the Plasmodium type that can cause malaria).
  • Target nucleic acids fall within the scope of templates in PCR terminology.
  • the template is more general and, in some cases, may be from any biological or synthetic DNA sequence.
  • the reaction mixture may include a reagent necessary for performing nucleic acid amplification, and the reaction mixture may further include a reporter for generating a detectable signal, and the reporter may detect the presence or absence of the signal, thereby indicating expansion. Whether the product is present.
  • the intensity of the detectable signal can be proportional to the amount of amplified product. In some cases, when the amplification product is generated from a different type of nucleic acid than the originally amplified target nucleic acid, the intensity of the detectable signal can be compared to the amount of the initially amplified target nucleic acid. example.
  • reagents necessary for the two reactions may further include a reporter that produces a detectable signal, which is detectable
  • the signal indicates the presence of the amplified DNA product and/or the amplified target RNA.
  • the intensity of the detectable signal can be proportional to the amount of amplified DNA product and/or amplified original target RNA.
  • Real-time amplification can also be performed by using a reporter, including real-time PCR for DNA amplification.
  • the reporter agent can be linked to the nucleic acid, including the amplification product, by covalent or non-covalent means.
  • non-covalent ways include ionic interactions, van der Waals forces, hydrophobic interactions, hydrogen bonding, and combinations thereof.
  • the reporter agent can bind to the initial reactant and the change in reporter level can be used to detect the amplification product.
  • the reporter agent can be detectable (or undetectable) only as the nucleic acid amplification proceeds.
  • optically active dyes eg, fluorescent dyes
  • Non-limiting examples of dyes include SYBR Green, SYBR Blue, DAPI, Propidium Iodine, Hoeste, SYBR Gold, Ethidium Bromide, Acridine, Protopantin, Acridine Orange, Acridine Yellow, Fluorescent Fluorcoumanin, ellipticine, daunorubicin, chloroquine, mitomycin D, chromomycin, homidium, fucomycin, ruthenium polypyridyl, Anqu Anthramycin, phenanthridine and acridine, ethidium bromide, propidium iodide, hexidium iodide, dihydrogen ingot, ethidium homodimer-1 and ethidium homodimerization Body-2, ethidium monoazide and ACMA, Hoechst 33258, Hoechst 33342, Hoechst 34580, DAPI, acridine orange, 7-AAD, actino
  • the reporter agent can be a sequence-specific oligonucleotide probe that is optically active upon hybridization with the amplification product. Due to the sequence-specific binding of the probe to the amplified product, the use of oligonucleotide probes can increase the specificity and sensitivity of the assay.
  • the probe can be attached to any of the optically active reporters (eg, dyes) described herein, and can also include a quencher capable of blocking the optical activity of the associated dye.
  • Non-limiting examples of probes that can be used as reporters include TaqMan probes, TaqMan Tamara probes, TaqMan MGB probes or Lion probes.
  • the reporter agent can be an RNA oligonucleotide probe comprising optical A reactive dye (eg, a fluorescent dye) and a quencher located adjacent to the probe. The close proximity of the dye to the quencher blocks the optical activity of the dye.
  • the probe can bind to the target sequence to be amplified. Once the exonuclease activity of the DNA polymerase cleaves the probe during amplification, the quencher is separated from the dye and the free dye regains its optical activity, which can then be detected.
  • the reporter agent can be a molecular beacon.
  • Molecular beacons include, for example, a quencher attached to one end of an oligonucleotide in a hairpin conformation. At the other end of the oligonucleotide is an optically active dye, such as a fluorescent dye. In the hairpin configuration, the optically active dye and quencher are sufficiently close together that the quencher is capable of blocking the optical activity of the dye. However, upon hybridization with the amplification product, the oligonucleotide is in a linear conformation and hybridizes to the target sequence on the amplification product.
  • Linearization of the oligonucleotide results in separation of the optically active dye from the quencher, thereby allowing optical activity to recover and can be detected.
  • the sequence specificity of the molecular beacon to the target sequence on the amplified product can improve the specificity and sensitivity of the assay.
  • the reporter agent can be a radioactive material.
  • radioactive materials include 14 C, 123 I, 124 I, 125 I, 131 I, Tc99m, 35 S, and 3 H.
  • the reporter agent can be an enzyme capable of producing a detectable signal.
  • the detectable signal can be produced by the activity of the enzyme on its substrate, or by the activity of the enzyme on a particular substrate with multiple substrates.
  • Non-limiting examples of enzymes useful as reported agents include alkaline phosphatase, horseradish peroxidase, I 2 - galactosidase, alkaline phosphatase, [beta] -galactosidase, and acetylcholinesterase firefly Photozyme.
  • lysing agent any suitable cleavage agent known in the art can be used, including commercially available lysing agents.
  • lysing agents include Tris-HCl, EDTA, detergents (eg, Triton X-100, SDS), lysozyme, glucosase, protease E, viral endolysin, exolysin ), zymolose, lyticase, proteinase K, endolysin and lysin from bacteriophage, endolysin from bacteriophage PM2, from B.
  • subtilis phage PBSX Lysin endolysin from Lactobacillus prophage Lj928, Lj965, phage 15 Phiadh, from pneumonia chain
  • the buffer may comprise a lysing agent (eg, a lysis buffer).
  • a lysis buffer is sodium hydroxide (NaOH).
  • nucleic acid amplification reaction Any type of nucleic acid amplification reaction known in the art can be used to amplify the target nucleic acid and generate an amplification product.
  • the amplification of the nucleic acid can be linear, exponential or a combination thereof.
  • the amplification can be emulsion based or not emulsion based.
  • Non-limiting examples of nucleic acid amplification methods include reverse transcription, primer extension, polymerase chain reaction, ligase chain reaction, helicase-dependent amplification, asymmetric amplification, rolling circle amplification, and multiple displacement amplification (MDA) ).
  • the amplification product can be DNA.
  • DNA can be obtained by reverse transcription of RNA and subsequent DNA amplification can be utilized to generate an amplified DNA product.
  • the amplified DNA product can indicate the presence of a target RNA in a biological sample.
  • any DNA amplification method known in the art can be used.
  • Non-limiting examples of DNA amplification methods include polymerase chain reaction (PCR), variants of PCR (eg, real-time PCR, allele-specific PCR, assembly PCR, asymmetric PCR, digital PCR, emulsion PCR, etc.), Helicase-dependent PCR, nested PCR, hot-start PCR, reverse PCR, methylation-specific PCR, miniprimer PCR, multiplex PCR, nested PCR, overlap-extension PCR, thermal asymmetric interleaving PCR (thermal asymmetric interlaced PCR), descending PCR) and ligase chain reaction (LCR).
  • PCR polymerase chain reaction
  • variants of PCR eg, real-time PCR, allele-specific PCR, assembly PCR, asymmetric PCR, digital PCR, emulsion PCR, etc.
  • Helicase-dependent PCR eg., nested PCR, hot-start PCR, reverse PCR, methylation-specific PCR, miniprimer
  • a primer set for a target nucleic acid can be used to perform nucleic acid Amplification reaction.
  • Primer sets typically contain one or more primers.
  • a primer set can comprise about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more primers.
  • the primer set can comprise primers for different amplification products or different nucleic acid amplification reactions.
  • a primer set can comprise a first primer and a second primer complementary to a nucleic acid strand product, the first primer being necessary for generating a first strand of a nucleic acid product complementary to at least a portion of the target nucleic acid, and the second primer is for generating a nucleic acid Essential for the second strand of at least a portion of the complementary nucleic acid product of the first strand of the product.
  • a primer set can be directed to a target RNA.
  • the primer set can comprise a first primer that can be used to generate a first strand of a nucleic acid product that is complementary to at least a portion of the target RNA.
  • the first strand of the nucleic acid product can be DNA.
  • the primer set can also comprise a second primer that can be used to generate a second strand of the nucleic acid product that is complementary to at least a portion of the first strand of the nucleic acid product.
  • the second strand of the nucleic acid product can be a strand of a nucleic acid (eg, DNA) product that is complementary to the DNA strand produced from the RNA template.
  • a nucleic acid eg, DNA
  • Any suitable number of primer sets can be used if desired. For example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more primer sets can be used. When multiple primer sets are used, one or more primer sets may each correspond to a particular nucleic acid amplification reaction or amplification product.
  • a DNA polymerase is used. Any suitable DNA polymerase can be used, including commercially available DNA polymerases.
  • DNA polymerase generally refers to an enzyme that is capable of incorporating a nucleotide into a DNA strand by template binding.
  • Non-limiting examples of DNA polymerase include Taq polymerase, Tth polymerase, Tli polymerase, Pfu polymerase, VENT polymerase, DEEPVENT polymerase, EX-Taq polymerase, LA-Taq polymerase, Expand polymerase, Sso Polymerase, Poc polymerase, Pab polymerase, Mth polymerase, Pho polymerase, ES4 polymerase, Tru polymerase, Tac polymerase, Tne polymerase, Tma polymerase, Tih polymerase, Tfi polymerase, Platinum Taq polymerization Enzyme, Hi-Fi polymerase, Tbr polymerase, Tfl polymerase, Pfutubo polymerase, Pyrobest polymerase, Pwo polymerase, KOD polymerase, Bst polymerase, Sac polymerase, Klenow fragment, and variants thereof, modifications Products and derivatives.
  • a hot start polymerase it may take 2 minutes to 10 minutes at
  • a reverse transcriptase is used. Any suitable reverse transcriptase can be used.
  • Reverse transcriptase generally refers to an enzyme capable of incorporating a nucleotide into a DNA strand upon binding to an RNA template.
  • Non-limiting examples of reverse transcriptases include HIV-1 reverse transcriptase, M-MLV reverse transcriptase, AMV reverse transcriptase, telomerase reverse transcriptase, and variants, modified products and derivatives thereof.
  • the reaction vessel can include a body for containing a reaction mixture, the body can include an inner surface, an outer surface, an open end, and a closed end, and the reaction mixture can be added to the reaction vessel via the open end.
  • the reaction vessel can also include a lid that can be in contact with the body at the open end to close the reaction vessel.
  • the lid is attached to the reaction vessel and remains attached to the reaction vessel when the container is opened and closed.
  • the lid can be separated therefrom when the reaction vessel is opened.
  • the fluid liquid or gas
  • the reaction vessel can be sealed (eg, gas sealed).
  • the reaction vessels described herein can have a variety of sizes, shapes, and weights.
  • the reaction vessel can be a circular or elliptical tube (eg, a PCR tube).
  • the reaction vessel can be rectangular, square, diamond, circular, elliptical or triangular.
  • the reaction vessel may be of a regular shape or an irregular shape.
  • the closed end of the reaction vessel can have a tapered, rounded or flat surface.
  • types of reaction vessels include tubes, wells, capillaries, cartridges, dishes, centrifuge tubes, or pipette tips.
  • the reaction vessel can be constructed from any suitable material, non-limiting examples of which include glass, metal, plastic, and combinations thereof.
  • the reaction vessel may be made of an optically clear material or a translucent material, for example, which may allow optical signals to pass through and/or penetrate from the reaction vessel.
  • the reaction vessel is made of a material that is filterable or non-filterable optical signals.
  • the reaction vessel may be made of a clarified material that may allow viewing of the interior of the reaction vessel by a detection device The situation.
  • the reaction vessel can have any suitable volume, for example, the volume of the reaction vessel can be adapted to hold at least about 0.2 milliliters (mL) or 0.5 mL of the reaction mixture.
  • the reaction vessel may be adapted to hold at least about 0.05 mL, 0.1 mL, 0.15 mL, 0.2 mL, 0.25 mL, 0.3 mL, 0.35 mL, 0.4 mL, 0.45 mL, 0.5 mL, 0.55 mL, 0.6 mL, 0.65 mL, 0.7 mL, 0.75 mL, 0.8 mL, 0.85 mL, 0.9 mL, 0.95 mL, 1 mL, 1.5 mL, 2 mL, 2.5 mL, 3 mL, 3.5 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 15 mL, 20 mL, 25m
  • the reaction vessel can be adapted to hold up to about 0.05 mL, 0.1 mL, 0.15 mL, 0.2 mL, 0.25 mL, 0.3 mL, 0.35 mL, 0.4 mL, 0.45 mL, 0.5 mL, 0.55 mL, 0.6 mL, 0.65 mL, 0.7 mL, 0.75 mL, 0.8 mL, 0.85 mL, 0.9 mL, 0.95 mL, 1 mL, 1.5 mL, 2 mL, 2.5 mL, 3 mL, 3.5 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL 10mL, 15mL, 20mL, 25mL, 30mL, 35mL, 40mL, 45mL, 50mL, 55mL, 60mL, 65mL, 70mL, 75mL, 80mL, 90mL,
  • the reaction vessel can be adapted to hold about 0.05 mL, 0.1 mL, 0.15 mL, 0.2 mL, 0.25 mL, 0.3 mL, 0.35 mL, 0.4 mL, 0.45 mL, 0.5 mL, 0.55 mL, 0.6.
  • the reaction vessel can have any suitable height, for example, the reaction vessel can have a height of at least about 0.25 centimeters (cm), at least about 0.5 cm, at least about 0.75 cm, at least about 1 cm, at least about 1.25 cm, at least about 1.5 cm, at least about 1.75 cm, at least about 2 cm, at least about 2.25 cm, at least about 2.5 cm, at least about 2.75 cm, at least about 3 cm, at least about 3.25 cm, at least about 3.5 cm, at least about 3.75 cm, at least about 4 cm, At least about 4.25 cm, at least about 4.5 cm, at least about 4.75 cm, at least about 5 cm, at least about 6 cm, at least about 7 cm, at least about 8 cm, at least about 9 cm, or at least about 10 cm.
  • cm centimeters
  • the height of the reaction vessel can be up to about 0.25 centimeters (cm), up to about 0.5 cm, up to about 0.75 cm, up to about 1 cm, up to about 1.25 cm, up to about 1.5 cm, up to about 1.75 Cm, up to about 2 cm, up to about 2.25 cm, up to about 2.5 cm, up to about 2.75 cm, up to about 3 cm, up to about 3.25 cm, up to about 3.5 cm, up to about 3.75 cm, up to about 4 cm, up to about 4.25 cm, up to About 4.5 cm, up to about 4.75 cm, up to about 5 cm, up to about 6 cm, up to about 7 cm, up to about 8 cm, up to about 9 cm, or up to about 10 cm.
  • cm centimeters
  • the height of the reaction vessel can be about 0.25 centimeters (cm), about 0.5 cm, about 0.75 cm, about 1 cm, about 1.25 cm, about 1.5 cm, about 1.75 cm, about 2 cm, about 2.25. Cm, about 2.5 cm, about 2.75 cm, about 3 cm, about 3.25 cm, about 3.5 cm, about 3.75 cm, about 4 cm, about 4.25 cm, about 4.5 cm, about 4.75 cm, about 5 cm, about 6 cm, about 7 cm, about 8 cm, about 9 cm or about 10 cm.
  • cm centimeters
  • the reaction vessel can have any suitable cross-sectional area.
  • the reaction vessel can have a cross-sectional area of at least about 0.1 square centimeters (cm 2 ), at least about 0.2 cm 2 , at least about 0.5 cm 2 , at least about 0.75.
  • cm 2 about at least 1cm 2, at least about 1.25cm 2, at least about 1.5cm 2, at least about 1.75cm 2, at least about 2cm 2, at least about 2.25cm 2, at least about 2.5cm 2, at least about 2.75cm 2, at least about 3cm 2, at least about 3.25cm 2, at least about 3.5cm 2, at least about 3.75cm 2, at least about 4cm 2, at least about 4.25cm 2, at least about 4.5cm 2, at least about 4.75cm 2, or at least about 5cm 2.
  • the reaction vessel has a cross-sectional area of at most about 0.25 square centimeters (cm 2 ), at most about 0.5 cm 2 , at most about 0.75 cm 2 , at most about 1 cm 2 , at most about 1.25 cm 2 , at most about 1.5cm 2, up to about 1.75cm 2, up to about 2cm 2, up to about 2.25cm 2, up to about 2.5cm 2, up to about 2.75cm 2, up to about 3cm 2, up to about 3.25cm 2, up to about 3.5cm 2 , up to about 3.75cm 2, at most about 4cm 2, up to about 4.25cm 2, up to about 4.5cm 2, up to about 4.75cm 2, or up to about 5cm 2.
  • the reaction vessel has a cross-sectional area of about 0.25 square centimeters (cm 2 ), about 0.5 cm 2 , about 0.75 cm 2 , about 1 cm 2 , about 1.25 cm 2 , about 1.5 cm 2 , about 1.75cm 2, about 2cm 2, about 2.25cm 2, about 2.5cm 2, about 2.75cm 2, about 3cm 2, about 3.25cm 2, about 3.5cm 2, about 3.75cm 2, about 4cm 2, about 4.25cm 2 , about 4.5cm 2, of about 4.75cm 2, or about 5cm 2.
  • the reaction vessel is part of an array of reaction vessels.
  • the array of reaction vessels can be used to automate methods and/or process multiple samples simultaneously.
  • the array can include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more reaction vessels.
  • the reaction vessel portion of the array of reaction vessels can also be accessed separately by the fluid treatment device such that the fluid treatment device can correctly identify the reaction vessel and dispense the appropriate fluid material into the reaction vessel.
  • the fluid treatment device can be used to automatically add fluid material to the reaction vessel.
  • the sample receiving unit of the present application may include one or more thermostat modules.
  • the thermostat module can be maintained at a substantially constant temperature, for example, plus or minus 5 ° C, 4 ° C, 3 ° C, 2 ° C, 1.2 ° C, 1 ° C, 0.7 ° C, 0.5 ° C, 0.3 ° C, near the set temperature, 0.1 ° C, 0.05 ° C, 0.01 ° C, 0.005 ° C or 0.001 ° C.
  • the temperature of the thermostat module may be a target temperature (ie, a temperature at which a chemical reaction proceeds) or a temperature slightly higher (when heated) or slightly lower (when cooled) than a target temperature. degree.
  • the thermostat module can be appropriately biased.
  • the temperature of the thermostatic module when heating or raising the temperature of the reaction mixture, can be set to be about 0.1 ° C higher than the target temperature, about 0.2 ° C higher, about 0.3 ° C higher, about 0.4 ° C higher, and about 0.5 ° C higher. , about 0.6 ° C high, about 0.7 ° C high, about 0.8 ° C high, about 0.9 ° C high, about 1.0 ° C high, about 1.1 ° C high, about 1.2 ° C high, about 1.3 ° C high, about 1.4 ° C high or about 1.5 ° C high. .
  • the temperature of the thermostatic module can be set to be about 0.1 ° C lower than the target temperature, about 0.2 ° C lower, about 0.3 ° C lower, about 0.4 ° C lower, about 0.5 ° C lower, It is about 0.6 ° C lower, about 0.7 ° C lower, about 0.8 ° C lower, about 0.9 ° C lower, about 1.0 ° C lower, about 1.1 ° C lower, about 1.2 ° C lower, about 1.3 ° C lower, about 1.4 ° C lower, or about 1.5 ° C lower.
  • the temperature of the thermostat module can range from about 80 ° C to about 110 ° C, from about 90 ° C to about 110 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 97 ° C, such as from about 92 ° C to about 95 ° C. .
  • the thermostat module has a temperature of at least about 80 ° C, at least about 81 ° C, at least about 82 ° C, at least about 83 ° C, at least about 84 ° C, at least about 85 ° C, at least about 86 ° C, at least About 87 ° C, at least about 88 ° C, at least about 89 ° C, at least about 90 ° C, at least about 91 ° C, at least about 92 ° C, at least about 93 ° C, at least about 94 ° C, at least about 95 ° C, at least about 96 ° C, at least About 97 ° C, at least about 98 ° C, at least about 99 ° C or at least about 100 ° C.
  • the temperature of the thermostat module is from about 30 ° C to about 80 ° C, from about 35 ° C to about 75 ° C, from about 35 ° C to about 72 ° C, from about 40 ° C to about 65 ° C, from about 45 ° C to About 60 ° C, about 50 ° C to about 60 ° C, for example, at least about 35 ° C, at least about 36 ° C, at least about 37 ° C, at least about 38 ° C, at least about 39 ° C, at least about 40 ° C, at least about 41 ° C, at least About 42 ° C, at least about 43 ° C, at least about 44 ° C, at least about 45 ° C, at least about 46 ° C, at least about 47 ° C, at least about 48 ° C, at least about 49 ° C, at least about 50 ° C, at least about 51 ° C, at least About 52 ° C, at least about 53 ° C, at least about 54 ° C, at least about
  • the contact time of the reaction mixture with the thermostat module can be controlled.
  • the contact time and offset can be determined by experiments, or can be derived by digital thermodynamic model, and then verified and modified by experiments. For example, such biasing is often used in conventional PCR instruments that use TEC to adjust temperature.
  • the thermostat module can be maintained at a specific temperature throughout the chemical reaction, or the temperature of a certain thermostat module can be changed from one temperature level to another during the thermal cycle.
  • the thermostat module can be prepared using a metal (eg, copper), a metal alloy, silica gel, and/or plastic.
  • the device of the present application may include a thermally insulating component.
  • the thermal insulation component can be located between the at least two thermostatic modules, and/or the thermal insulation component can thermally isolate the sample receiving unit from the outside environment.
  • the thermally insulating component can be used to prevent or reduce heat transfer, such as heat transfer between thermostatic modules or between a thermostatic module and a reaction vessel.
  • Exemplary thermal insulation materials include, but are not limited to, air or other gases, vacuum, plastic foam, glass, rubber, fabric, fiberglass, or combinations thereof.
  • the thermostat module can include holes, slots, projections, depressions, or other shapes that are designed to match the reaction vessel.
  • the matching can improve thermal contact between the thermostatic module and the reaction vessel.
  • a detection unit can be used to detect the generated signal in a chemical reaction, such as a nucleic acid amplification reaction.
  • the resulting reaction can be detected without removing the reaction vessel from the apparatus of the present application. signal.
  • the detection device can detect an amplified product (eg, an amplified DNA product, an amplified RNA product), and the like.
  • the amplified product (including amplified RNA) can be detected by any suitable method.
  • the appropriate detection method can be determined depending on the specific amplification product, the reaction vessel used, other reagents in the reaction mixture, whether or not a reporter is contained in the reaction mixture, and the like.
  • Non-limiting examples of detection methods include optical detection, spectroscopic detection, electrostatic detection, and electrochemical detection.
  • Optical detection methods include, but are not limited to, fluorescence measurements and UV-visible absorption.
  • Spectroscopic detection methods include, but are not limited to, spectroscopic analysis, nuclear magnetic resonance (NMR) spectroscopy, and far infrared spectroscopy.
  • Electrostatic detection methods include, but are not limited to, gel-based techniques such as gel electrophoresis.
  • Electrochemical detection methods include, but are not limited to, electrochemical detection of amplified products after separation of the amplified products using high performance liquid chromatography.
  • the detection unit can be fixed to the device of the present application, which can be driven by a separate motor or share one or more motors with other components.
  • the detection unit may include an image sensor capable of detecting an optical signal.
  • the image sensor can include a charge coupled device (CCD) sensor.
  • the image sensor may comprise an active pixel image sensor (APS), such as a CMOS or NMOS sensor.
  • APS active pixel image sensor
  • the detection unit may include a laser sensor.
  • the detection unit comprises a photodiode, such as an avalanche photodiode.
  • the detection unit may include a photomultiplier tube (PMT).
  • the sensor may include one or more sensors, which may be the same or different types of sensors.
  • the sensor can detect an optical signal from the reaction mixture.
  • the optical signal can be a fluorescent signal or other luminescent signal from the reaction mixture.
  • the reaction mixture can produce the optical signal in response to stimulation of the excitation light.
  • the light source that generates the excitation light may include a lamp (eg, an incandescent lamp, a halogen lamp, a fluorescent lamp, a gas discharge lamp, an arc lamp, or an LED lamp).
  • the light source can include a laser source.
  • the light source can produce light having a specific wavelength or range of wavelengths, such as UV.
  • the light source can include filtering means for controlling the wavelength of the output.
  • the excitation light may come from a plurality of light sources of the same or different types, and they may be used alone or in combination.
  • the light source can be built into the device or system of the present application or can be independent of the device or system of the present application. And exist.
  • the excitation light can be absorbed by the reaction mixture and the reaction mixture can emit light.
  • the light emitted by the reaction mixture may be the same as or different from the wavelength of the absorbed light.
  • the optical signal can be obtained by reflection of light from a light source.
  • the light can illuminate through the reaction mixture and the detection unit can detect light passing through the reaction mixture.
  • An optical pathway can be provided between the reaction vessel and the detection unit.
  • a signal from the reaction mixture can pass through the optical pathway to the detection unit.
  • the optical channel may be a line of sight between the reaction vessel and the detection unit.
  • one or more optical elements eg, lenses, mirrors, prisms, diffusers, concentrators, filters, optical fibers, etc.
  • the device of the present application can include a housing that is substantially opaque and can include at least one optical pathway, and wherein the nucleic acid amplification reaction can be detected in the detection unit by an optical pathway on the housing The situation that was carried out.
  • information regarding the presence and/or amount of the target can be output to the recipient.
  • This information about the amplified product can be output by any suitable means.
  • the information about the progress and/or results of the reaction can be provided in real time while the chemical reaction (e.g., nucleic acid amplification reaction) is being performed.
  • the information can be output to the recipient after the reaction is over.
  • the apparatus or system of the present application can include a low voltage power source.
  • the voltage of the power source may be, for example, less than or equal to about 60V, 50V, 48V, 40V, 30V, 24V, 20V, 18V, 16V, 15V, 14V, 13V, 12V, 11V, 10V, 9V, 8V, 7V, 6V. 5V, 4V, 3V, 2V or 1V, this voltage is sufficient for thermal cycling and/or detection of reaction products.
  • the device or system of the present application can also be operatively coupled to an energy storage device.
  • the energy storage device can be a battery case or a charger.
  • the battery may be a lithium battery, a lead acid battery, a valve regulated lead acid battery, a nickel cadmium (NiCd) battery, a nickel zinc (NiZn) battery, or a nickel hydrogen battery. (NiMH) and so on.
  • the energy storage device can be part of the device or system of the present application. In certain embodiments, the energy storage device can be provided in a housing of the device of the present application. In certain embodiments, the energy storage device can be removed from the device or system of the present application.
  • the application also provides a computer control system that is programmed to implement the methods and/or aspects of the present application.
  • the computer control system can be integrated as part of the apparatus or system of the present application.
  • the computer control system can be configured to control the operation of the thermal cycler of the present application and collect data.
  • the computer control system can regulate various aspects of the apparatus of the present application, such as temperature, ramp rate, fade time, number of cycles, and data collection of the thermostat module.
  • the computer control system can be an electronic device of a user or computer system that can remotely control the apparatus or system of the present application.
  • the computer control system can include a central processing unit, which can be a single or multiple core processor.
  • the computer control system can also include a memory or memory address, an electronic storage unit (eg, a hard disk), and/or an interactive interface for communicating with the outside world.
  • the memory, storage unit, interactive interface, and any peripheral devices can be in operative communication with a central processing unit.
  • the user prepares a PCR reaction tube and adds 40 ⁇ L of the reaction mixture thereto.
  • the reaction mixture was cycled between 94 degrees Celsius and 64 degrees Celsius by the apparatus of the present application.
  • the volumetric specific heat capacity of the PCR reaction mixture was 4.2 J/cm 3 *K, and the heat capacity of the 40 ⁇ L reaction mixture was 0.168 J/K, that is, 0.168 J/°C.
  • the apparatus shown in Figure 1 is used in which the first thermostat module and the second thermostat module are both made of copper because of the higher thermal conductivity and specific heat capacity of the copper.
  • the volume of the first thermostat module is about 2.82 cm 3
  • the specific heat capacity of copper is 3.45 J/cm 3 *K, so the heat capacity of the first thermostat module is 9.73 J/° C.
  • the heat capacity ratio between the PCR reaction mixture and the first thermostat module is 0.0173, that is, the temperature of the first thermostat module drops by 0.017 ° C for every 1 degree Celsius increase in the temperature of the PCR reaction mixture.
  • the temperature of the first thermostat module will drop by 0.52 ° C.
  • the temperature change of the thermostat module during heating and cooling is small.
  • in order to heat the PCR reaction mixture to 94 ° C it is only necessary to preheat the first thermostat module to about 94.52 ° C. This small fluctuation in temperature facilitates precise temperature control.
  • the energy output of the components used for heating is greatly reduced.
  • the energy consumed (increasing the temperature from 64 ° C to 94 ° C) was only 5.04 J for each PCR cycle. If the reaction is set to 10 seconds/cycle, the power output requirement for the heating element is about 0.5W.
  • the lower temperature (64 ° C) thermostat module acts as a heat sink.
  • the PCR reaction mixture transfers 5.04 J of thermal energy to the cooling thermostat module, which will raise the temperature of the cooling thermostat module by 0.52 ° C. Accordingly, excess heat energy on the cooling thermostat module will dissipate to the surrounding In the environment.
  • the cooling thermostat module needs to be set to the desired temperature (eg, about 63.48 °C) before the first thermal cycle begins. Once the cycle begins, there is no need to heat the cooling module.
  • the temperature of the cooling thermostat module can be kept constant only by, for example, a fan.
  • TEC thermoelectric cooling fins
  • Thermal energy is transferred from the heating/cooling element (TEC) via the transfer module.
  • the volume of the transfer module is 0.15 cm 3 .
  • the material of the TEC is aluminum and the volumetric specific heat capacity is 2.4 J/cm 3 * ° C.
  • the module has a heat capacity of 0.36 J/° C.
  • the combined heat capacity of the module and the PCR reaction mixture was 0.53 J/° C.
  • the module requires 15.9 J of thermal energy in combination with the PCR reaction mixture.
  • the power output requirement of the heating element is about 3.2 W.
  • the electric power required to transfer 15.9 J of heat from the thermal energy transfer module to the heatsink will be greater than 3.2 W.
  • the thermal conductivity of the module is also considered (ie, the thermal conductivity is not considered to be infinite)
  • the power requirements will be higher in order to generate a sufficiently large temperature gradient to transfer energy. Therefore, when using the TEC, additional power is required to make the TEC temperature much higher than the preset target temperature so that the set temperature can be reached more quickly.
  • the invention of the present application has lower requirements for energy consumption and higher efficiency. This is due to the fact that during the temperature change, it is only necessary to raise the temperature of the reaction mixture without increasing the temperature of both the reaction mixture and the module. On the other hand, the module temperature remains constant throughout the entire reaction cycle, rather than heat transfer only during the heating step.
  • the present invention provides an apparatus and method and system for performing a chemical reaction that is capable of achieving fast, low power consumption, portability, ease of operation, and/or accurate effects, and thus, the industrialization of the present invention Very useful.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Clinical Laboratory Science (AREA)
  • Immunology (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)

Abstract

L'invention porte sur un appareil (200), sur un procédé et sur un système pour mettre en oeuvre une réaction chimique. L'invention peut être utilisée de manière spécifique pour mettre en oeuvre une amplification d'acide nucléique ou une autre réaction chimique, avec une exigence de température différente, pour le test et l'analyse d'échantillons. L'invention est rapide, a une faible consommation d'énergie, est pratique et facile à utiliser, et/ou fournit un résultat précis.
PCT/CN2017/095000 2016-08-08 2017-07-28 Appareil, procédé et système pour la mise en oeuvre d'une réaction chimique WO2018028447A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610638913.5A CN106367336B (zh) 2016-08-08 2016-08-08 用于进行化学反应的装置、方法和系统
CN201610638913.5 2016-08-08

Publications (1)

Publication Number Publication Date
WO2018028447A1 true WO2018028447A1 (fr) 2018-02-15

Family

ID=57878282

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/095000 WO2018028447A1 (fr) 2016-08-08 2017-07-28 Appareil, procédé et système pour la mise en oeuvre d'une réaction chimique

Country Status (2)

Country Link
CN (1) CN106367336B (fr)
WO (1) WO2018028447A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106367336B (zh) * 2016-08-08 2020-03-03 皮卡（上海）生物科技有限公司 用于进行化学反应的装置、方法和系统
CN110157590A (zh) * 2018-02-13 2019-08-23 光鼎生物科技（江苏）有限公司 热循环仪

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110244522A1 (en) * 2010-03-30 2011-10-06 Tae Seok Seo Rotational pcr equipment and pcr method using the same
CN102791847A (zh) * 2010-01-12 2012-11-21 阿赫姆生物系统公司 三阶段热对流装置及其用途
CN103820316A (zh) * 2014-03-09 2014-05-28 北京工业大学 基于旋转式微流控芯片的实时荧光pcr检测系统
CN104046556A (zh) * 2013-03-13 2014-09-17 精工爱普生株式会社 核酸扩增反应装置和核酸扩增方法
CN105296344A (zh) * 2014-05-28 2016-02-03 精工爱普生株式会社 核酸扩增反应装置
CN106367336A (zh) * 2016-08-08 2017-02-01 皮卡（上海）生物科技有限公司 用于进行化学反应的装置、方法和系统

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102791847A (zh) * 2010-01-12 2012-11-21 阿赫姆生物系统公司 三阶段热对流装置及其用途
US20110244522A1 (en) * 2010-03-30 2011-10-06 Tae Seok Seo Rotational pcr equipment and pcr method using the same
CN104046556A (zh) * 2013-03-13 2014-09-17 精工爱普生株式会社 核酸扩增反应装置和核酸扩增方法
CN103820316A (zh) * 2014-03-09 2014-05-28 北京工业大学 基于旋转式微流控芯片的实时荧光pcr检测系统
CN105296344A (zh) * 2014-05-28 2016-02-03 精工爱普生株式会社 核酸扩增反应装置
CN106367336A (zh) * 2016-08-08 2017-02-01 皮卡（上海）生物科技有限公司 用于进行化学反应的装置、方法和系统

Also Published As

Publication number Publication date
CN106367336A (zh) 2017-02-01
CN106367336B (zh) 2020-03-03

Similar Documents

Publication Publication Date Title
Lobato et al. Recombinase polymerase amplification: Basics, applications and recent advances
Li et al. All-in-one microfluidic device for on-site diagnosis of pathogens based on an integrated continuous flow PCR and electrophoresis biochip
CN103403521B (zh) 流体向心装置
CN106520517B (zh) 一种便携式微流控芯片lamp可视化检测仪及其检测方法
Li et al. Recent advancements in nucleic acid detection with microfluidic chip for molecular diagnostics
TW201723168A (zh) 用於進行化學反應的設備和方法
ES2809566T3 (es) Métodos y sistemas para la amplificación de ácidos nucleicos
US10487301B2 (en) Reaction tube for nucleic acid amplification capable of controlling liquid circulation path
KR102256757B1 (ko) 중합효소 연쇄반응 시스템
WO2019143812A1 (fr) Procédés de dosage pour déceler la présence de micro-organismes
Chang et al. Detection of viruses directly from the fresh leaves of a Phalaenopsis orchid using a microfluidic system
US20180080063A1 (en) Apparatus and methods for conducting chemical reactions
US20190185909A1 (en) Methods and systems for analyzing nucleic acids
EP1371419A1 (fr) Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon
WO2018028447A1 (fr) Appareil, procédé et système pour la mise en oeuvre d'une réaction chimique
US20190184402A1 (en) Methods and systems for analyzing nucleic acids
KR102039785B1 (ko) 핵산증폭 반응의 온도조절 방법 및 그 용도
US20170128947A1 (en) Devices and methods for monitoring and quantifying nucleic acid amplification
US20230407382A1 (en) Methods and systems for sample analysis
WO2016106717A1 (fr) Appareils et procédés permettant de réaliser des réactions chimiques
KR102113957B1 (ko) 어류양식장 휴대용 유전자 추출 및 등온증폭 pcr 분석장치
US20220033884A1 (en) Methods and systems for identification of spinal muscular atrophy
CN206256055U (zh) 一种在微流控芯片内进行核酸扩增的装置
CN112359099A (zh) 用于核酸扩增的方法和系统
US20230302446A1 (en) Cartridge for detecting target analyte

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17838577

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17838577

Country of ref document: EP

Kind code of ref document: A1