WO2022147604A1 - Machine portable automatique de détection d'acides nucléiques en une étape - Google Patents

Machine portable automatique de détection d'acides nucléiques en une étape Download PDF

Info

Publication number
WO2022147604A1
WO2022147604A1 PCT/CA2021/000002 CA2021000002W WO2022147604A1 WO 2022147604 A1 WO2022147604 A1 WO 2022147604A1 CA 2021000002 W CA2021000002 W CA 2021000002W WO 2022147604 A1 WO2022147604 A1 WO 2022147604A1
Authority
WO
WIPO (PCT)
Prior art keywords
sample
chamber
tube
piston rod
pcr
Prior art date
Application number
PCT/CA2021/000002
Other languages
English (en)
Inventor
Zonghua LIU
Original Assignee
Molarray Research Inc.
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 Molarray Research Inc. filed Critical Molarray Research Inc.
Priority to PCT/CA2021/000002 priority Critical patent/WO2022147604A1/fr
Publication of WO2022147604A1 publication Critical patent/WO2022147604A1/fr

Links

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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/54—Supports specially adapted for pipettes and burettes
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0621—Control of the sequence of chambers filled or emptied
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/16—Reagents, handling or storing thereof
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0672—Integrated piercing tool
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/087—Multiple sequential chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B01L2300/1827—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using resistive heater
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0478—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure pistons
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00346—Heating or cooling arrangements
    • G01N2035/00356—Holding samples at elevated temperature (incubation)
    • G01N2035/00366—Several different temperatures used

Definitions

  • TITLE PORTABLE AUTOMATIC ONE-STEP NUCLEIC ACID DETECTION MACHINE
  • the present invention relates generally to equipment for medical’ diagnosis for respiratory diseases and in specific to a point-of-care-testing (POCT) device for POCT.
  • POCT point-of-care-testing
  • PCR Polymerase chain reaction
  • POCT point-of- care-testing
  • swab nasopharyngeal, throat, etc.
  • PCR machines are not generally automated in testing facilities, although there are some available technologies that are designed for commercial research and testing facilities.
  • nucleic acid extraction kits are in high demand and short supply. Extraction-free techniques that have developed during this time, have become more prevalent, eliminating the need for a separate nucleic acid extraction step. However, these techniques still require a separate transferring step, from sample collection to the testing facility, where the qPCR is run. Once again, this includes possible exposure and cross-contamination risk as samples must be opened to be put into the qPCR machine manually.
  • the present invention is a portable automatic one-step nucleic acid detection machine for disease diagnosis, especially for diagnosis of respiratory viruses such as SARS-Co-V2/Flu A/Flu B and can be used in point-of-care-testing facilities.
  • the point-of-care-testing machine comprises a sample processing unit, a PCR module and a heat block module.
  • the machine effectively combines the extraction-free sampling process with qPCR machine.
  • Patient samples can be taken and added directly to the sample processing unit where the extraction-free process takes place followed by heating and mixing the sample with various reagents and the qPCR reaction. This further allows the diagnosis of pathogens and eliminates the need for transporting the sample to a testing facility, and also removes an extra hazard level by keeping the potentially infected sample in a closed system from post-collection all the way to qPCR results and diagnosis.
  • An important feature of the invention is that it allows multiple stepwise processing of the sample to be done sequentially all in one closed system automatically, with minimal manual intervention. This is accomplished by having sequential chambers in the sample processing unit, each initially sealed off by a film. This allows buffers and solutions to be combined in a specific order in the sampling process, without having to open and expose the system.
  • the sample processing unit comprises of chambers separated by aluminum films or similar type of materials. Mainly, the machine functions by having a cylinder run through the top chamber, cutting through the films to begin mixing the solutions. A piston rod runs through the chambers all the way down to the bottom chamber, which is the PCR tube. The chambers are connected continuously in a vertical line one after another and each chamber receives a chemical for sample processing.
  • a driving mechanism is provided to hold and drive the piston rod through the chambers and deliver a positive predetermined pressure to puncture each of the films and function as a syringe to facilitate mixing a predetermined amount of chemicals in the chambers.
  • a piston ring is provided on the bottom of the piston rod to allow the sealing of the tube at the final step to prevent evaporation of the reaction during PCR. Additionally, specific airways are designed in this device to allow the release of air pressure as the cylinder and piston are depressed, to facilitate movement but also to prevent as much cross-contamination . as possible with the outside environment as well.
  • the machine is configured to facilitate the automation of the whole process. Ball screws allow the consumable sample tube to be locked into place on the machine.
  • the machine has a set of plates to control the movement of the cylinder and the piston rod. A cylinder plate controls the depression of the cylinder while a rod plate controls the depression of the piston rod. In this way, the machine can be programmed to control each separately, facilitating automatic processing.
  • the present invention provides a closed vessel reaction, drastically limiting crosscontamination opportunities.
  • the invention is portable and can be used bench-side or bedside, allowing diagnostic testing in facilities other than a testing laboratory. The whole process also takes place in a little over an hour, which is significant in diagnostic testing, as test results usually take a minimum of several days to come back from testing laboratories.
  • [20]Another object of the present invention is to integrate sample collection and processing with qPCR analysis into one convenient device in order to achieve quick, bedside diagnostic testing of pathogens normally diagnosed by qPCR.
  • Another object of the present invention is to minimize the risk of contamination, especially with pathogens such as SARS-CoV2 that have especially high risk for those handling the samples. This also minimizes transport time and overall turnaround time for diagnosis of such diseases.
  • Another object of the present invention is to automate the testing process to provide more consistent results but also to make the system as simple as possible with minimal manual intervention. This should effectively increase both safety and reliability of the invention.
  • Another object of the present invention is that it is easy to use, as it involves only two points of manual intervention: adding the sample, as well as releasing the tube when the reaction is finished.
  • This automation allows more consistent testing as well as provides easy clean-up from a closed system.
  • This invention also benefits in the turnaround time of testing; as current diagnostic testing often takes a minimum of several days. Quicker diagnostic test results can lead to a prompter treatment regime for the patient.
  • FIG. 1A is a perspective view showing the point of care testing machine (POCT) of the present invention
  • FIG. 1 B is a perspective view of the corresponding sample processing unit of the present invention.
  • FIG. 2 is a cross-sectional view showing the main parts of the sample processing unit of the POCT machine of the present invention
  • FIG. 3 is a cross-sectional view showing the arrangement of the chemical reagents in the chambers of the processing unit according to the present invention
  • FIG. 4 is a cross-sectional close-up view showing the cylinder and the piston rod of the sample processing unit of the present invention
  • FIG. 5 is a cross-sectional view showing the tube and the piston rod and the airways created in the chambers according to the present invention
  • FIG. 6 is a cross sectional view of a partial of the sample processing unit showing the dents and the O-rings sealed the air way to prevent of contamination of the air according to one aspect of the present invention
  • FIG. 7 is a cross sectional view showing the dimensions of various parts of the sample processing unit, according to one embodiment of the present invention
  • FIG. 8A is a perspective view of the POCT machine
  • FIG. 8B is a perspective front view of the POCT machine
  • FIG. 9 is a perspective view showing the rear side of the POCT machine.
  • FIG. 10 is a perspective partially transparent view of FIG.9 showing the PCR tubes inside the Heat block and PCR module of the POCT machine;
  • FIG. 11 A is a cross-sectional view of the sample processing unit showing the step 1 of the process by adding the sample into the tube;
  • FIG. 11 B is a cross-sectional view of the sample processing unit showing the step 1 of the process according to FIG. 11 A;
  • FIG. 12 is a perspective view of the POCT machine showing the step 2 of the process by setting the pistons and tubes inside the machine
  • FIG. 13A is a perspective view of the POCT machine showing the operation of the machine in the step 3 and how the machine automatically drives the cylinder and pistons into the tubes, pierces the films and adds the chemicals;
  • FIG. 13B is a cross sectional view of the sample processing unit according to FIG. 13A;
  • FIG. 13C is a cross sectional view of the sample processing unit according to FIG. 13A and 13B;
  • FIG. 14A is a cross-sectional view of the sample processing unit showing the step 4 of the process by cutting aluminum films and the piston rod going up and down mix the solution;
  • FIG. 14B is a cross sectional view of the sample processing unit showing the step 4 of the process according to FIG. 14A;
  • FIG. 15A is a cross-sectional view showing the step 5 of the process by piston rod piercing aluminum film to add Lysis Buffer and mix the solution again;
  • FIG. 15B is a cross-sectional view showing the step 5 of the process according to FIG. 15A;
  • FIG. 16A is a cross-sectional view showing the step 6 of the process measuring 10ul liquid
  • FIG. 16B is a cross-sectional view showing the step 6 of the process according to FIG. 16A;
  • FIG. 17 is a cross-sectional close-up view of the bottom part of the sample processing unit showing the step 7 of the process in which the piston rod cut the film to let the air exit and push the liquid into the PCR tube;
  • FIG. 18 is a cross-sectional view of the sample processing unit in which the piston rod drives back to seal the air way by the O-ring;
  • FIG. 19 is a cross-sectional view of the sample processing unit showing the step 8 of the process to start ⁇ the PCR reaction;
  • FIG. 20 is a perspective view showing the optical module for the PCR module, of the POCT machine.
  • FIG. 21 is a cross-sectional view showing the PCR machine of the present invention
  • FIG. 22 is a perspective view of the POCT machine showing the Step 9 of the process in which the reaction is finished and the PCR tubes can be taken out from the machine.
  • the Point of Care Testing(POCT) machine 200 comprises a sample processing unit 100, a heat block 220, a PCR module 210 and a computer program to control the extraction free sample process.
  • Plurality of sample processing unit 100 can be installed into the POCT machine 200.
  • the system and method of the invention provides for rapid and simplified processing of a sample.
  • the POCT machine 200 has a frame 211 , which comprises of all the necessary components to process a sample to obtain a desired target compound.
  • the frame 211 is made of a rigid material, such as metal, like aluminum, that is both light but also can be machined to high tolerance and is sturdy enough. All the components of the invention can also be made by injection molding.
  • the machine 200 effectively combines the extraction-free sampling process with the PCR machine 210.
  • the POCT 200 further comprises a data storage component (“DSC”), which stores data related to sample processing or reagents used.
  • DSC data storage component
  • the computer program of the system is configured to provide instructions for operating various parts of the machine 200 and the PCR module 210 per protocol contained on the DSC. Data can be uploaded to or downloaded from the DSC through conventional wireless or wired technologies.
  • the whole process takes place in a sample processing unit 100. Patients’ samples can be taken and added directly to the unit 100 where the extraction- free process takes place followed by the qPCR reaction.
  • the sample processing unit 100 has a specific design configured to allow multiple stepwise processing of the sample to be done sequentially all in one closed system automatically, with minimal manual intervention. This is accomplished by having sequential chambers in the sample collector, each initially sealed off by a film. This allows multiple solutions to be combined in a specific order in the process, without having to open and expose the system.
  • the sample processing unit 100 is a sealed tube 10 and has a pipette tip geometry.
  • the preferred tube 10 configuration is substantially cylindrical so that the flow vectors during operation are substantially straight.
  • the tube 10 is dimensioned to fit onto the heat block of the machine 220.
  • Suitable materials are not limited for the tube and include plastics (such as polyethylene, polypropylene, and polystyrene), glass and stainless steel.
  • the tube 10 comprises of a plurality of chambers (tubes) connected continuously in a vertical line one after another and separated by aluminum films to receive various chemicals for sample processing.
  • the current tube 10 is configured to hold four types of chemicals with specific volumes, as shown in FIG. 3. However, tubes with other number of chambers and any desired volumes can be designed for a particular protocol.
  • the current tube 10 contains the following chemical reagents from the top to the bottom of the tube 10: Buffer (150 pi) 30, Lysis Buffer (50 pi) 31 , Reagent (30 mI) 32 and Enzyme (1 OmI) 33.
  • TE Buffer is filled into the second chamber 3b, which is separated from the first chamber 3a by aluminum film 6.
  • the aluminum films are selected from common qPCR sealing films.
  • the third chamber 2a is filled with Lysis Buffer 31 and is separated from the second chamber 3b by aluminum film 7.
  • the fourth chamber 2b is filled with reagent 32 and is separated from third chamber 2a by aluminum film 8.
  • the fifth chamber or PCR tube 1 is filled with enzyme 33 and is separated from the fourth chamber 2b, by aluminum film 9.
  • the sample processing unit 100 further includes a cylinder 5 which drives through the tube 10.
  • a piston rod 12 drives through the middle of the cylinder 5 and punctures the films further down in the chambers, as well as functions as a type of syringe to facilitate mixing of the solutions.
  • the piston 11 with the piston rod 12 are configured to drive in a vertical position upwards and downwards inside the cylinder 5 all the way through the tube 10 to apply positive pressure, where the release of the contents is effected through puncture of the aluminum films or negative pressure to release air from the cylinder 5.
  • the piston rod 12 has a sharp tip 13 configured to pierce the films by a predetermined pressure.
  • the sample processing unit 100 functions by having the cylinder 5 run through the tube 10, cutting through the films 6, 7, 8 and 9 to begin mixing the solutions.
  • An important feature of the invention is that it allows multiple stepwise processing of the sample to be done sequentially all in one closed system automatically, with minimal manual intervention. This is accomplished by having sequential chambers in the sample processing unit 100, each initially sealed off by a film. This allows solutions to be combined in a specific order in the process, without having to open and expose the system
  • FIGs. 4 to 6 show how the cylinder 5 and piston rod 12 move through the tube 10 and facilitate the processes.
  • the machine 200 can automatically run a program to drive the piston 11 , piston rod 12 and the cylinder 5 up and down, and run PCR module according to the workflow.
  • the sizing of the piston rod 12 and the corresponded tube 10 are calculated to ensure the proper volumes are attained. In this preferred embodiment the volume of the cylinder 5 minus the volume of the core equals to 10mI.
  • the piston rod 12 is fully depressed with the cylinder 5
  • all the solutions are mixed with the final reagents and flow into the PCR tube 1 .
  • a set of dents 14 and 16 interact with slits on the side of the cylinder 5 to allow for air to escape and release pressure while the contents of the first and second chamber 3a and 3b are initially mixed.
  • the piston rod 12 moves up and down to suck air out and push the liquid in for mixing.
  • specific airways are designed in this invention to allow the release of air pressure as the cylinder 5 and piston rod 12 are depressed, to facilitate movement but also to prevent as much cross-contamination as possible with the outside environment as well.
  • An air passage is created by the dents 14 and 16 and a gap 15 for the air to go in and out to balance the air pressure inside the chambers.
  • the dents 14 and 16 and the gap 15 are designed to create a path for air 28.
  • a top O-ring 18 acts to seal the gap 15 between the cylinder 5 and the piston rod 12 allowing it to act as a syringe for mixing purposes.
  • a piston ring 17 allows the sealing of the first and second chambers 3a and 3b at the final step of the process to prevent evaporation of the reaction during PCR.
  • the piston 11 moves to a lower position.
  • the dents 14 and 16 move away from the O-ring 18, thereby the O-ring 18 seals the air way and the liquid is sealed inside the chamber, thereby the vapor cannot escape out to contaminate the air.
  • FIG. 7 shows the dimensions of the parts of the sample processing unit 100 in the preferred embodiment.
  • the unit 100 can be designed in various sizes and dimension for different protocols.
  • FIGs. 8A to 10 illustrate the POCT machine 200.
  • the machine 200 comprises of a vertical frame 211 .
  • a set of horizontal plates 201 and 202 are movable mounted on the front side of the frame 211 and move up and down on the linear railings 207 and 208.
  • the upper plate 201 is configured to hold the piston rod 12 and the lower plate 202 is configured to hold the cylinder 5 of the processing unit 100.
  • the cylinder 5 and piston rod 12 are locked into the plates 201 and 202 by a plurality of ball screws.
  • Trollo motors 203 and 204 are provided onto the bottom of the machine 200 to drive the upper plate 201 and the lower plate 202 upwards and downwards.
  • the motors 203 and 204 rotate a set of ball screws 205 and 206 to drive the plates 203 and 204 upwards and downwards, thereby move the cylinder 5 and the piston rod 12.
  • the operation of each plate 201 and 202 is controlled separately by each motor.
  • the motor 203 operates the upper plate 201 and the motor 204 operates the lower plate 202.
  • the motors are in communication with a processor or controller that controls the movement of the cylinder 5.
  • the processor or controller can send instructions to activate or stop the vertical motion of the cylinder 5.
  • the machine itself facilitates the automation of the whole process.
  • the lower plate 202 controls the depression of the cylinder 5 while the upper plate 201 controls the depression of the piston rod 12. In this way, the machine 200 can be programmed to control each of the plates 201 and 202 separately, facilitating automatic processing.
  • the PCR module 210 is installed on the rear side of the machine. The machine works in conjunction with a heat block unit 220.
  • the heat block 220 is in a location above the PCR machine 210, allowing the heat inactivation of the sample before PCR, without affecting the PCR reagents themselves, which are sensitive to heat.
  • the tubes 10 and the pistons 12 are set into the machine 200 and locked in place by a plurality of plungers 209 provided on the machine 200. In this way, sample collection all the way to PCR output can be automated by the invention.
  • the machine 200 is preferably constructed of a relatively stiff, strong and sterilisable material that may be assembled to provide structural support to the various units of the machine 200 and to be able to operate a fully automated sample testing system.
  • the overall process flow of the invention is depicted in FIGs. 11 A to 22.
  • step 1 as shown in FIG. 11 A and 11 B a sample is added to the first chamber 3a and flows into the second chamber 3b for mixing with the TE buffer 30.
  • Four types of chemicals are preset in the tube 10 and sealed and separated with the aluminum films 6, 7, 8 and 9 respectively.
  • the chemicals consist of: TE Buffer 30 (150 pi); Lysis Buffer 31 (50pl); Reagent 32(30pl) and Enzyme 33 (1 pg powder).
  • step 2 and 3 as shown in FIG. 12 to 13C, the tube 10 and the piston 12 are set into the machine 200 and locked in place by a plurality of plungers 209 provided on the machine 200.
  • the machine 200 starts to operate and the motors drive the two plates 201 and 202 to move the piston 11 and rod 12 downwards into the tubes 10.
  • the tubes 10 are fixed on heat blocks 220 and the machine 200 drives piston 11 and piston rod 12 in the tubesl 0 while the tube 10 is heated by the heat block 220 up to 95°C.
  • the heat block 220 is in a location above the PCR machine 210, allowing the heat inactivation of the sample before PCR, without affecting the PCR reagents themselves, which are sensitive to heat.
  • the cylinder 5 and the piston rod 12 drive further downwards and push to punch the aluminum film 6, thereby the sample- flows into the second chamber 3b where it mixes with TE buffer 30.
  • the piston rod has a sharp tip 13 configured to pierce the aluminum films by a predetermined force, which will be automatically applied by the machine.
  • the bottom space of the second chamber 3b will retain 10mI of liquid. Meanwhile, piston rod 12 will pierce aluminum film 7 to open an air channel that leads from the PCR tube 1 , through the gap 15 between the piston rod 12 and the cylinder 5 allowing for air to escape, so that the build of air pressure does not prevent the proper flow of the solutions into the PCR tube 1.
  • the machine can automatically run a program to heat, drive the cylinder 5 and piston rod 12 up and down, and run PCR according to the workflow.
  • step 4 the piston rod 12 drives up and down to mix the liquid while heated by the heat block 220.
  • the air can enter and exit into the chamber to balance the pressure.
  • the exit of the air can let the reagent (150 mI TE Buffer) enter the chamber.
  • the piston rod 12 continuously drives up and down 30 minutes to mix the liquid.
  • the piston rod 12 releases a set amount of the liquid in the fourth chamber 2b where the liquid will be isolated.
  • 10 mI liquid is measured and released into the fourth chamber 2b.
  • the piston rod 12 drives upwards and releases O-ring 18 to seal the top air way, thereby the liquid will be isolated in the fourth chamber 2b.
  • the fourth chamber 2b fits snugly into the heat block 210 and PCR machine 210.
  • the piston rod 12 pierce aluminum film 8 and let the air exit the chamber through an air channel 28 and pushes 10 pi liquid and 30 pi reagent 32 into the fourth chamber 2b for further reaction.
  • the cylinder 5 and piston rod 12 are designed to measure out various volumes of reagents needed in the process and to mix them all together. It is to be understood that the sizing of the tube10 in the sample processing unit 100 can be adjusted to fit different protocols. For example, the PCR reaction tube 1 could be bigger or smaller depending on the reaction volume required, or depending on the size of the PCR machine. One element that might increase the flexibility of this invention would be to allow the switching of the reagents in the tubes for further applications.
  • the piston rod 12 pierce aluminum film 9 and pushes 10 pi liquid into the PCR tube 1 for the reaction to take place via the PCR machine 210.
  • the PCR tube 1 has enzymes 33 for the reaction process.
  • FIG. 20 depicts the PCR 210 module of the Machine 200.
  • the PCR module 210 is placed beneath the heat block 220 of the machine. Therefore, the present invention eliminates the separate transferring step, from sample collection to the testing facility where the PCR is run, which may include possible exposure and cross-contamination risk as samples must be opened to be put into the PCR machine manually.
  • the present disclosure is an implementation of a PCR machine 210 with an optical module and depicts the excitation light system of the first channel.
  • the beam of light coming from the LED 410 is aligned through the collimator lens 400, then passed through an optical filter 510 to filter out the stray light.
  • the filtered beam passes through the focusing lens 500 to be coupled, and the coupled beams subsequently pass through the launch fiber 600 to be re-coupled with the other beams.
  • the re-coupled beam passes through the light guide 700 and should pass evenly into 4 launch fiber 650, 651 , 652, 653 to the PCR tube 800.
  • the emission lights from the reaction tube would pass to the receiving fibers 900 to the optical filters 810.
  • the stray excitation and emission lights would be filtered out by the filter and then coupled and guided into the tapered light guide 910. These lights will converge onto the photodiode 920 for signal processing.
  • the signal acquisition processes of the second, third and fourth channel following the other LEDs 420- 430- 440, optical filters 520-530-540 and optical filters 820- 830- 840 follow the same process as the first channel.
  • FIG. 21 shows a cross sectional view of the POCT machine 200 and the arrangement of the sample processing unit 100 of the invention.
  • FIG. 22 shows the last step of the procedure in the POCT machine 200.
  • the reaction procedure is finished and the tubes 10 are ready to take out.
  • the point- of-care-testing machine 200 can be used in PCR machines for analyzing any nucleic acid containing sample for any purpose, including clinical testing for various infectious diseases including bacterial or viral. This machine can be configured to run on a laboratory benchtop, or similar environments and the results are typically available in less than 1 hour.
  • a computer program for use with the machine 200 herein includes computer readable instructions thereon for operating the machine.
  • the computer program can include one or more instructions to cause the system to operate components of the system.
  • the machine has a processor, such as a microprocessor, configured to control functions of various components of the system.
  • The, processor can be configured to receive data about a sample to be analyzed, e.g., from a sample reader, which may be a barcode reader, an optical character reader, or an RFID scanner (radio frequency tag reader).
  • a processor is configured to accept user instructions from an input.
  • Processor can be also configured to communicate with a display, so that, for example, information about an analysis is transmitted to the display and thereby communicated to a user of the system.
  • Processor can be optionally further configured to transmit results of an analysis to an output device such as a printer, a display or a combination thereof.
  • Processor can be still further optionally connected via a communication interface such as a network interface to a computer network.
  • the machine 200 can further comprise a data storage medium configured to receive data from one or more of the processor.
  • Processor can be further configured to control various aspects of sample preparation and diagnosis such as operation of the piston and cylinder, operation of the heat block and the motors and the solution dispense operations in the chambers to carry out various dispense operations on respective sample, fluids and reagents in the chambers.
  • the device may be configured to carry out operation in a single location, such as a laboratory setting, or may be portable so that it can accompany, e.g., a physician, or other healthcare professional, who may visit patients at different locations.
  • the machine 200 is typically provided with a power-cord so that it can accept AC power from a mains supply or generator.
  • the machine 200 may also be configured to operate by using one or more batteries and therefore is also typically equipped with a battery recharging system, and various warning devices that alert a user if battery power is becoming too low to reliably initiate or complete a diagnostic analysis.
  • the present invention is easy to use, as it involves only two points of manual intervention: adding the sample, as well as releasing the tube when the reaction is finished.
  • Extraction-free qPCR analysis of patient samples is still a relatively new technique, mostly fast-tracked in light of the SARS-CoV2 pandemic that has stretched global supplies for RNA extraction kits to the limit.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Clinical Laboratory Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Hematology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un dispositif permettant le traitement automatique des acides nucléiques d'un échantillon, puis la réaction et l'analyse qPCR, le tout dans une seule cuve de réaction. La technique élimine le besoin d'extraction d'acide nucléique, et peut générer des résultats de diagnostic rapides avec une manipulation manuelle minimale. Après avoir ajouté l'échantillon, la machine peut se charger du traitement de l'échantillon en utilisant un type de piston de seringue pour mélanger successivement les réactifs entre eux. Un bloc thermique permet l'inactivation des échantillons et la PCR ultérieure a également lieu dans la même cuve de réaction. De cette manière, l'ensemble du traitement de l'échantillon et la PCR peuvent avoir lieu dans un système fermé, renforçant la sécurité et la fiabilité des tests.
PCT/CA2021/000002 2021-01-06 2021-01-06 Machine portable automatique de détection d'acides nucléiques en une étape WO2022147604A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CA2021/000002 WO2022147604A1 (fr) 2021-01-06 2021-01-06 Machine portable automatique de détection d'acides nucléiques en une étape

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CA2021/000002 WO2022147604A1 (fr) 2021-01-06 2021-01-06 Machine portable automatique de détection d'acides nucléiques en une étape

Publications (1)

Publication Number Publication Date
WO2022147604A1 true WO2022147604A1 (fr) 2022-07-14

Family

ID=82357029

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2021/000002 WO2022147604A1 (fr) 2021-01-06 2021-01-06 Machine portable automatique de détection d'acides nucléiques en une étape

Country Status (1)

Country Link
WO (1) WO2022147604A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116202817A (zh) * 2022-11-23 2023-06-02 安徽沙丰新材料有限公司 橡胶用水性硬脂酸钙分散液取样装置

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006121997A2 (fr) * 2005-05-09 2006-11-16 Idaho Technology, Inc. Analyse biologique autonome
WO2009009144A2 (fr) * 2007-07-12 2009-01-15 Smiths Detection Inc. Appareil de préparation d'échantillon

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006121997A2 (fr) * 2005-05-09 2006-11-16 Idaho Technology, Inc. Analyse biologique autonome
WO2009009144A2 (fr) * 2007-07-12 2009-01-15 Smiths Detection Inc. Appareil de préparation d'échantillon

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116202817A (zh) * 2022-11-23 2023-06-02 安徽沙丰新材料有限公司 橡胶用水性硬脂酸钙分散液取样装置

Similar Documents

Publication Publication Date Title
JP6838127B2 (ja) 統合された移送モジュールを有する試験カートリッジ
JP6937774B2 (ja) 生物学的アッセイを実行するためのシステムおよび方法
CN108220155B (zh) 用于分子诊断的系统和方法
CN111386465A (zh) 诊断测试系统和方法
US20110215118A1 (en) Dispensing cylinder, large capacity dispensing device and method of using large capacity dispensing device
KR20170024827A (ko) 미세유로 필름 반응기, 핵산 추출 모듈 및 qPCR 반응조성물 모듈이 구비된 qPCR 카트리지 및 이를 이용한 고속 qPCR 시스템
US20220258168A1 (en) Diagnostic cartridge for microfluidics and on-site molecular diagnosis system including same
WO2022147604A1 (fr) Machine portable automatique de détection d'acides nucléiques en une étape
US6200781B1 (en) Apparatus, system and method for automated execution and analysis of biological and chemical reactions
EP2677321A1 (fr) Dispositif intégré pour l'hybridation d'acides nucléiques et analyses immunologiques
KR102513521B1 (ko) 현장형 자동 진단 시스템을 위한 카트리지
US20230381770A1 (en) Sample Preparation Cartridge and System
EP4382915A1 (fr) Dispositif de point d'intervention pour la mise en oeuvre d'une variété de réactions biochimiques différentes
US12024739B1 (en) Method and apparatus for rapid analysis of a biological sample
JP2024510383A (ja) Ｄｎａまたはｒｎａ増幅装置とその手法
WO2021198476A1 (fr) Diagnostics de point d'intervention contrôlés par pression

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: 21916680

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: 21916680

Country of ref document: EP

Kind code of ref document: A1