Infectious disease testing system

Abstract

An infectious disease testing system (1) for testing for infectious diseases such as COVID-19 infection. The system includes an ultrasonic transducer (49) that generates ultrasonic waves to lyse cells in a biological sample. The system (1) includes a controller that controls the ultrasonic transducer (49) to vibrate at an optimal frequency for lysing the cells, a PCR device (16) that receives and amplifies DNA from the sample, and a detection device (70) that detects the presence of the infectious disease in the amplified DNA and provides an output indicating whether a detection sequence (70) detects the presence of the infectious disease in the amplified DNA.

Description

Technical Field

[0001] (Cross - reference to related applications) This application claims the benefit of each of the following priorities and incorporates the entire contents herein by reference: European Patent Application No. 20177685.3 filed on June 1, 2020, European Patent Application No. 20200852.0 filed on October 8, 2020, European Patent Application No. 20214228.7 filed on December 15, 2020, and International Patent Application No. PCT / GB2021 / 050822 filed on April 1, 2021.

[0002] The present invention relates to an infectious disease testing system for performing tests on infectious diseases including, but not limited to, COVID - 19 infection. More specifically, the present invention relates to an infectious disease testing system for testing viral infectious diseases using the polymerase chain reaction (PCR) process, including, but not limited to, testing for SARS - CoV - 2 viral infection.

Background Art

[0003] Advances in technology in the medical field have improved the efficiency of diagnostic methods and diagnostic devices. The testing time has been significantly shortened, and at the same time, reliable results can be obtained. There are various testing methods for detecting all kinds of infectious diseases. In the testing of viral infectious diseases, PCR (polymerase chain reaction) has proven to be the most reliable method. Similar to other methods, the PCR method has also evolved as a more time - efficient and cost - effective method while maintaining high reliability.

[0004] PCR is a technique that uses two matching strands in DNA to amplify the target DNA sequence from just a few samples to billions of copies, and as a result, the DNA samples are analyzed by gel electrophoresis that separates them according to their size.

[0005] [[ID=2五]] Conventional polymerase chain reaction (PCR): Conventional PCR tests are composed of the following 3 - 4 steps.

[0006] 1. Cell lysis and nucleic acid (DNA / RNA) extraction: After collecting a patient sample from the nose (nasopharyngeal swab) or throat (oropharyngeal swab), mix the sample with the elution buffer. Filter the eluate to remove large particles (hair, skin fragments, etc.). Pour the filtered solution into the dissolution tank.

[0007] This provides a gateway for cell lysis, disrupting the lipid bilayer of cells in a sample and extracting cellular components, including DNA / RNA.

[0008] Cell lysis is carried out chemically, electromechanically, or a combination of both. This process involves extracting components and filtering the solution to separate nucleic acids (DNA / RNA) from other cellular components. This DNA / RNA can then proceed to the next step.

[0009] 2. Reverse transcription (RT): This step is only necessary if the nucleic acid is RNA and not DNA.

[0010] In this process, an enzyme called reverse transcriptase is introduced into a PCR solution containing RNA, and a complementary DNA (cDNA) sequence is created from the RNA at a temperature of 40-50°C. Since PCR requires DNA or cDNA, the reverse transcription step is performed before any PCR-related actions.

[0011] 3. Polymerase chain reaction (PCR) The principle of PCR is the same regardless of the type of DNA sample. PCR requires five core materials: the DNA sample, primers, DNA bases, polymerase enzyme, and buffer to ensure the appropriate conditions for the reaction.

[0012] PCR involves a heating and cooling process called thermal cycling. Thermal cycling consists of three steps: denaturation, annealing, and extension.

[0013] Denaturation begins by heating the reaction solution to 95°C to 100°C. This high temperature is necessary to separate double-stranded DNA or cDNA into single strands.

[0014] Annealing is the process of binding primers to denatured strands of sample DNA or cDNA. This process requires a temperature of 55°C to 62°C. Once this temperature is reached, the annealing stage begins, in which the primers bind to a single strand.

[0015] Once the primer binds, polymerase binds and extends the primer along the length of the single strand, raising the temperature to approximately 72°C to create new double-stranded DNA.

[0016] To obtain optimal results, the thermal cycle should be repeated 20 to 40 times, depending on the number of base pairs required for the test, ensuring that the desired temperature is achieved at each stage.

[0017] 4. Gel electrophoresis After PCR is complete, the quantity and size of the generated DNA fragments can be confirmed using a method called electrophoresis. Because DNA is negatively charged, to separate them by size, the PCR-treated sample is placed on an agarose gel, and an electric current is passed through the gel to attract the negatively charged DNA to the opposite end. Larger DNA fragments encounter greater resistance in the solution and therefore do not move as much as smaller fragments in the same amount of time.

[0018] The test results are obtained by comparing the migration distance of DNA fragments with that of a known sample. During solution preparation, before the gel electrophoresis step, a fluorescent dye is added to identify the DNA band, and the length of the DNA can be determined from its position.

[0019] Fast PCR: High-speed PCR shortens the overall inspection time by performing thermal cycling in a shorter time (20 - 60 seconds per cycle) compared to conventional PCR. Additionally, high-speed PCR employs a real-time PCR process, an automated high-speed thermal cycling process that performs amplification and detection in a single step within a sealed reaction vessel. This process can significantly reduce the risk of infection. High-speed PCR uses fluorescence spectroscopy to detect simultaneously with the thermal cycle of PCR.

[0020] When testing for viruses (RNA), high-speed RT-PCR adds one step to the overall test. That additional step is a reverse transcription step to create cDNA from RNA before the above-mentioned PCR step.

[0021] Fluorescence spectroscopy: Fluorescence spectroscopy is used to shorten the overall test time as an alternative to gel electrophoresis. Fluorescence spectroscopy uses light to excite electrons within the molecules of a specific compound and cause them to emit light. The detector senses this light to measure the fluorescence, enabling the identification of molecules and molecular changes.

[0022] Due to the global pandemic of the SARS-CoV-2 virus (COVID-19 infection), the demand for virus test kits has increased rapidly. Also, conventional tests take 4 - 8 hours to complete, and even high-speed tests take more than 2 hours, so faster tests are being demanded.

[0023] In conventional virus tests, it is common to process a large number of samples simultaneously. However, because the time for each step (mainly PCR) is long, the waiting time until the results are obtained is long. In the high-speed PCR method, by shortening the thermal cycle time, a certain lead time is ensured compared to the conventional PCR method, and the overall test time is shortened to about 1 - 2 hours. However, even this test time is too long for useful large-scale high-speed testing for infectious diseases such as COVID-19.

[0024] There is a need for improved systems and devices for infectious disease testing that mitigate at least some of the problems outlined in this specification.

Summary of the Invention

[0025] Some configurations of infectious disease testing systems include the following. An ultrasonic treatment chamber configured to receive a biological sample to be tested for an infectious disease, an ultrasonic transducer configured to output ultrasonic waves in a frequency range of about 2800 kHz to about 3200 kHz to lyse cells from the biological sample within the ultrasonic treatment chamber, a controller having an AC driver configured to generate an alternating current drive signal at a predetermined frequency within the frequency range of about 2800 kHz to about 3200 kHz and configured to output the alternating current drive signal to drive the ultrasonic transducer, an effective power monitoring device configured to monitor the effective power used by the ultrasonic transducer when the ultrasonic transducer is driven by the alternating current drive signal, the effective power monitoring device being configured to provide a monitoring signal indicative of the effective power used by the ultrasonic transducer, a processor configured to control the AC driver and receive the monitoring signal from the effective power monitor, and when executed by the processor, the processor A. Controls the AC driver to output an alternating current drive signal to the ultrasonic transducer at a predetermined sweep frequency, B. Calculates the effective power being used by the ultrasonic transducer based on the monitoring signal, C. Controls the AC driver to modulate the alternating current drive signal to maximize the effective power used by the ultrasonic transducer, D. Saves a record of the maximum effective power used by the ultrasonic transducer and the sweep frequency of the alternating current drive signal in memory, E. After a predetermined number of iterations, repeats steps A through D for a predetermined number of iterations such that the sweep frequency increments from a sweep start frequency to a sweep end frequency, incrementing the sweep frequency in each iteration, F. From the records stored in memory, identify the optimal frequency of the AC drive signal, which is the sweep frequency of the AC drive signal at which the maximum active power is used by the ultrasonic transducer, and G. The system includes a memory that stores instructions for controlling an AC driver to output an AC drive signal to an ultrasonic transducer at an optimal frequency. The system further includes a polymerase chain reaction (PCR) apparatus configured to receive and amplify DNA from lysed cells of a biological sample, and an infectious disease detection device configured to detect the presence of an infectious disease in the amplified DNA and to provide an output indicating whether the infectious disease detection device has detected the presence of an infectious disease in the amplified DNA.

[0026] In some configurations, the active power monitor includes a current sensor to sense the drive current of the AC drive signal that drives the ultrasonic transducer, and the active power monitor is designed to provide a monitoring signal indicating the sensed drive current.

[0027] In some configurations, memory stores instructions that, when executed by the processor, cause the processor to increment the sweep frequency from a starting sweep frequency of 2800 kHz to an ending sweep frequency of 3200 kHz and repeat steps A through D.

[0028] In some configurations, memory stores instructions that, when executed by the processor, cause the processor to control an AC driver to output an AC drive signal to an ultrasonic transducer at a frequency shifted between 1% and 10% of the optimal frequency in step G.

[0029] In some configurations, the AC driver modulates the AC drive signal by pulse width modulation to maximize the active power used by the ultrasonic transducer.

[0030] In some configurations, memory stores instructions that, when executed by the processor, cause the processor to control an AC driver, alternately performing actions such as outputting an AC drive signal to the ultrasonic transducer at an optimal frequency during a first predetermined time, and not outputting an AC drive signal to the ultrasonic transducer during a second predetermined time.

[0031] In some configurations, memory stores instructions that, when executed by the processor, cause the processor to alternate between outputting AC drive signals and not outputting AC drive signals, according to an operating mode selected from the following:

[0032] [Table 1]

[0033] In some configurations, the system further comprises a heating device incorporating a heating recess that receives part of a PCR device, a movable support element, a first heating element supported by the support element, a second heating element supported by the support element at a position spaced apart from the first heating element (characterized by the support element being movable between a first position in which the first heating element is positioned closer to the heating recess than the second heating element and a second position in which the second heating element is positioned closer to the heating recess than the first heating element), and a motor configured to periodically move the support element between the first and second positions.

[0034] In some configurations, the heating device includes a temperature sensor configured to sense the temperature of the liquid in the PCR device located within a heating recess, and a controller configured to control the movement of first and second heating elements in response to the sensed temperature.

[0035] In some configurations, the controller is configured to control a first heating element to heat the liquid in the PCR instrument to substantially 45°C during the reverse transcriptase step.

[0036] In some configurations, during the PCR process, the controller is configured to control a first heating element to heat the liquid in the PCR instrument to substantially 55°C, control a second heating element to heat the liquid in the PCR instrument to substantially 95°C, and periodically move a support element between first and second positions so that the first and second heating elements circulate the temperature of the liquid in the PCR instrument between substantially 55°C and substantially 95°C.

[0037] In some configurations, the system further includes movable channels to selectively provide fluid channels between the sample chamber, sonication chamber, or PCR chamber, enabling the continuous transfer of at least a portion of the sample between the sample chamber, sonication chamber, and PCR chamber.

[0038] In some configurations, the system further includes a filtration configuration configured to filter the fluid flowing out of the movable channel. This filtration configuration comprises a first filter element with pores ranging from 2 μm to 30 μm in diameter.

[0039] In some configurations, the filtration system includes a second filter element superimposed on a first filter element. The pores of the second filter element range in diameter from 0.1 μm to 5 μm.

[0040] In some configurations, the system is a COVID-19 infection testing system, and this infection detection device is a SARS-CoV-2 virus detection device configured to detect the presence of the SARS-CoV-2 virus that causes COVID-19 infection in amplified DNA and to provide an output indicating whether or not the SARS-CoV-2 virus detection device detects the presence of COVID-19 infection in the amplified DNA.

[0041] Some configurations of infectious disease testing methods include: placing a biological sample to be tested for infectious disease into an ultrasonic processing chamber (this ultrasonic processing chamber is equipped with an ultrasonic transducer that outputs ultrasound in the frequency band of approximately 2800 kHz to approximately 3200 kHz and dissolves cells obtained from the biological sample within the ultrasonic processing chamber); generating an AC drive signal at a predetermined frequency within the frequency range of approximately 2800 kHz to approximately 3200 kHz using an AC driver, outputting this AC drive signal to the ultrasonic transducer to drive the ultrasonic transducer; monitoring the active power used by the ultrasonic transducer when the ultrasonic transducer is driven by the AC drive signal using an active power monitor (the active power monitor is characterized by providing a monitoring signal indicating the active power used by the ultrasonic transducer); and receiving the monitoring signal from the active power monitor with a processor, and the method further consists of the following.

[0042] A. The processor controls the AC driver to output an AC drive signal to the ultrasonic transducer at a predetermined sweep frequency. B. The processor calculates the active power used by the ultrasonic transducer based on the monitoring signal. C. The processor modulates the AC drive signal and controls the AC driver to maximize the active power used by the ultrasonic transducer. D. The processor stores in memory the maximum active power used by the ultrasonic transducer and the sweep frequency of the AC drive signal. E. After a predetermined number of iterations, steps A to D are repeated a predetermined number of times with the sweep frequency that is increased in each iteration, such that the sweep frequency increases from the sweep start frequency to the sweep end frequency. F. The processor identifies the optimal frequency of the AC drive signal, which is the sweep frequency of the AC drive signal at which the maximum active power is used by the ultrasonic transducer, from the records stored in memory, and G. A processor controls the AC driver to output an AC drive signal to the ultrasonic transducer at an optimal frequency. This method is characterized by receiving and amplifying DNA from lysed cells of a biological sample using a polymerase chain reaction (PCR) device, detecting the presence of an infectious disease in the amplified DNA using an infectious disease detection device, and providing an output indicating whether or not the infectious disease detection device has detected the presence of an infectious disease in the amplified DNA.

[0043] In some configurations, the method further includes using a current sensor to sense the drive current of an AC drive signal that drives an ultrasonic transducer, and using a current sensor to provide a monitoring signal indicating the sensed drive current.

[0044] In some configurations, this method further involves pulse-width modulation of the AC drive signal by an AC driver in order to maximize the active power used by the ultrasonic transducer.

[0045] In some configurations, the method further includes controlling an AC driver by a processor, which alternately outputs an AC drive signal to the ultrasonic transducer at an optimal frequency for a first predetermined time, and then does not output an AC drive signal to the ultrasonic transducer for a second predetermined time.

[0046] In some configurations, this method further includes controlling the AC driver alternately to output an AC drive signal and then to stop outputting an AC drive signal, according to an operating mode selected from the following:

[0047] [Table 2]

[0048] In some configurations, the method is a COVID-19 infection testing method, which includes detecting the presence of the SARS-CoV-2 virus that causes COVID-19 infection in amplified DNA using a SARS-CoV-2 virus detection device, and providing an output indicating whether or not the SARS-CoV-2 virus detection device detects the presence of COVID-19 infection in the amplified DNA. [Brief explanation of the drawing]

[0049] To make the present invention more easily understood, embodiments of the present invention will now be described by reference to the accompanying drawings.

[0050] [Figure 1] Figure 1 is a schematic perspective view of a system in a configuration that includes an assay apparatus in a configuration. [Figure 2] Figure 2 is a schematic diagram of an assay apparatus with a specific configuration. [Figure 3] Figure 3 is a schematic diagram showing a part of a system with an assay apparatus in a specific configuration. [Figure 4] Figure 4 is a schematic perspective view showing a portion of an assay apparatus in a specific configuration. [Figure 5] Figure 5 is a side view showing a part of the assay apparatus shown in Figure 4. [Figure 6] Figure 6 is an end view showing a part of the assay apparatus shown in Figure 4. [Figure 7] Figure 7 is a schematic diagram showing a portion of an assay apparatus in a particular configuration. [Figure 8] Figure 8 is a cross-sectional view of a part of the assay apparatus shown in Figure 7. [Figure 9] Figure 9 is a cross-sectional view of a part of the assay apparatus shown in Figure 7. [Figure 10] Figure 10 is a schematic diagram showing the components of some filtration configurations. [Figure 11] Figure 11 is a schematic diagram showing a portion of an assay apparatus in a particular configuration. [Figure 12]Figure 12 is a schematic diagram showing a piezoelectric transducer modeled as an RLC circuit. [Figure 13] Figure 13 is a graph of frequency versus logarithmic impedance for an RLC circuit. [Figure 14] Figure 14 is a graph of frequency versus logarithmic impedance showing the inductive and capacitive operating regions of a piezoelectric transducer. [Figure 15] Figure 15 is a flowchart illustrating the operation of a controller in a specific configuration. [Figure 16] Figure 16 is a partial perspective view of an assay apparatus in a particular configuration. [Figure 17] Figure 17 is a partial perspective view of an assay apparatus in a specific configuration. [Figure 18] Figure 18 is a partial perspective view of an assay apparatus in a specific configuration. [Figure 19] Figure 19 is a side view showing a part of the assay apparatus shown in Figure 18. [Figure 20] Figure 20 is an end view showing a part of the assay apparatus shown in Figure 18. [Figure 21] Figure 21 is a cross-sectional view showing part of the system in one configuration and part of the assay apparatus in another configuration. [Figure 22] Figure 22 is a perspective view showing a portion of the system in one configuration and a portion of the assay apparatus in another configuration. [Figure 23] Figure 23 is a side view showing a portion of an assay apparatus in a particular configuration. [Figure 24] Figure 24 is a perspective view showing part of a system in a particular configuration. [Modes for carrying out the invention]

[0051] Detailed explanation The aspects of this disclosure will be best understood by the following detailed description provided with the accompanying drawings. Note that, in accordance with standard industry practice, various features in the drawings are not depicted to scale. In fact, the dimensions of various features may be arbitrarily increased or decreased for the sake of clarity in the discussion.

[0052] The following disclosure provides many different embodiments or examples for carrying out various features of the subject matter provided. Specific examples of components, concentrations, uses, and arrangements are described below for the sake of brevity of this disclosure. Naturally, these are merely examples and are not intended to limit the scope. For example, the mounting of the first and second features in the following description may include embodiments in which the first and second features are mounted in direct contact, and may also include embodiments in which an additional feature may be positioned between the first and second features so that the first and second features do not have to be in direct contact. In addition, the disclosure may repeat reference numerals and / or letters in various examples. This repetition is for simplification and clarity and is not in itself intended to indicate relationships between the various embodiments and / or configurations discussed.

[0053] The following disclosures describe typical configurations or examples. Each example can be considered an embodiment, and references to “configuration” or “example” may be replaced with “embodiment” in this disclosure.

[0054] This disclosure establishes an improved embodiment of a rapid result diagnostic assay system designed for point-of-care (POC) and / or home use for infectious disease testing, particularly for SARS-CoV-2, which is known to cause COVID-19 infection.

[0055] Some configurations of assay devices and systems are for testing for any other infectious diseases caused by pathogens such as bacteria or viruses. In some configurations, assay devices and systems are for testing for infectious substances or diseases selected from the group including, but not limited to, influenza, coronavirus, measles, HIV, hepatitis, meningitis, tuberculosis, Epstein-Barr virus (glandular fever), yellow fever, malaria, norovirus, Zika virus infection, or anthrax.

[0056] In some configurations, the assay apparatus and system are for testing target samples in the form of saliva, sputum, or blood samples. In other configurations, the assay apparatus and system are for testing target samples collected from the user by nasopharyngeal or oropharyngeal swab.

[0057] Some configurations of the assay system comprise 13 main components, including an assay apparatus or pod containing various liquid chambers, a plunger column, flow direction gears, an ultrasonic chamber, a filtration array, PCR fins, PCR reagents, a PCR method, a thermal cycler, an infection detection device, a lid, a results reporting method, and a housing containing all necessary components for operating the pod.

[0058] Referring to Figure 1 of the attached drawings, System 1 for infectious disease testing is configured in this configuration to be used with a removable assay device 2, which is in the form of a single-use pod. In some configurations, System 1 is supplied separately from the assay device 2. In other configurations, System 1 is supplied in combination with the assay device 2. In yet another configuration, the assay device 2 is supplied without System 1 but is used in combination with System 1.

[0059] System 1 comprises a housing 3 that accommodates various components of System 1. In this configuration, the housing 3 has an opening 4 that is closed by a door 5. The door 5 is configured to move between an open position, as shown in Figure 1, and a closed position, where the door 5 closes the opening 4 of the housing 3. In this configuration, the door 5 is provided with a handle 6 to facilitate opening and closing by the user. In this configuration, the door 5 is provided to allow the user to open System 1 and insert the assay apparatus 2 into System 1, as is commonly indicated by arrow 7 in Figure 1. In other configurations, different access means are incorporated to allow the user to insert the assay apparatus 2 into System 1.

[0060] In this configuration, System 1 is a portable system. The enclosure 3 is compact so that System 1 can be easily carried and placed inconspicuously in a convenient location, such as adjacent to a building entrance door. The portable configuration of System 1 in some configurations allows System 1 to be easily transported to locations where infectious disease testing is needed. In some configurations, System 1 is configured to be powered by a battery or other low-power source so that System 1 can be used in remote locations without requiring a mains power supply. In other configurations, System 1 has a power input that is connected to a mains power supply to power System 1 and / or charge the battery in System 1.

[0061] System 1 includes a support base 8 provided at the base of the housing 3. The support base 8 has a surface on which the assay device 2 is placed. The support base 8 includes a plurality of guide members 9 arranged around the support base 8 to guide the assay device 2 into a predetermined position when the assay device 2 is inserted into System 1. In this arrangement, the support base 8 has a central opening 10 that is located beneath the assay device 2 when the assay device 2 is carried by the support base 8.

[0062] Referring next to Figure 2 of the attached drawings, in this arrangement, the assay apparatus 2 comprises a base 11, which has an enlarged lower end to provide stability to the assay apparatus 2 when the assay apparatus 2 is resting on the base 11. The assay apparatus 2 further comprises an assay apparatus housing 12 that houses the internal components of the assay apparatus 2, which will be described in more detail below. The assay apparatus housing 12 has an upper end 13 away from the base 11, which is configured to be open to provide access to the inside of the assay apparatus 2. A cover 14 is movably attached to the assay apparatus housing 12 so as to at least partially cover the upper end 13. The cover 14 has a central opening 15. The cover 14 will be described in more detail below.

[0063] The assay apparatus 2 includes a PCR device 16 that protrudes from one side of the assay apparatus 2. The PCR device 16 will be described in more detail below.

[0064] Next, referring to Figure 3 of the attached drawings, when the assay device 2 is inserted into the system 1, the assay device 2 is guided to a predetermined position on the support base 8 such that the PCR device 16 is at least partially received in the heating recess of the heating device 17, which will be described in detail below.

[0065] Assay apparatus 2 is located beneath a drive unit 18, which forms part of system 1. In this arrangement, the drive unit 18 comprises a drive element in the form of a plunger 19. This is configured such that the tip 20 of the plunger 19 moves outward by the drive unit 18 through an opening 15 in the cover 14 of assay apparatus 2, generally in the direction indicated by arrow 21, and engages with a piston 22 in assay apparatus 2. System 1 is configured to extend and retract the plunger 19 to move the piston element 22 during the operation of system 1.

[0066] System 1 includes a controller 23 that incorporates an arithmetic unit such as a microprocessor and memory. The controller 23 is configured to control the operation of System 1, as described below.

[0067] Referring here to Figures 4-6 of the attached drawings, the assay apparatus 2 comprises a body portion 24 that is elongated and defines at least one internal chamber. In this arrangement, the body portion 24 has sides defined by eight generally planar surfaces arranged such that the body portion 24 has an octagonal cross-section. However, it should be understood that in other arrangements, body portions with different shapes and different cross-sections may be incorporated.

[0068] In this configuration, the main body 24 defines multiple internal chambers. In this configuration, the main body 24 defines six internal chambers: a sample chamber 25, a washing chamber 26, a solvent chamber 27, a liquid reagent chamber 28, a dry reagent chamber 29, and a waste chamber 30. The main body 24 is also provided with a central opening 31.

[0069] The number of chambers in the assay apparatus can vary from 1 to a maximum of 10 depending on the configuration. In the configuration for the SARS-CoV-2 assay, assay apparatus 2 has 6 chambers.

[0070] One end of the main body portion 24 is provided with a protrusion 32, as shown in Figure 5. The protrusion 32 is provided with a plurality of openings 33, as shown in Figure 6. Each opening 33 provides a fluid communication path to one of the chambers 25 to 30.

[0071] Next, referring to Figure 7 of the attached drawings, the assay apparatus 2 includes a transfer device 34 that is movably mounted on the main body portion 24. The transfer device 34 includes a plunger column 35 that defines an elongated transfer chamber 36. In this arrangement, the plunger column 35 is an elongated, generally cylindrical column configured to be at least partially received within the central opening 31 of the assay apparatus body 24.

[0072] The plunger column 35 is the central part of the assay apparatus 2. The liquid contained within the assay apparatus 2 is moved and manipulated between various chambers as it progresses through all stages of preparation for PCR. The transfer chamber 36 contains a rubber plunger tip connected to the plunger arm contained within the housing of System 1. The liquid is drawn into the transfer chamber 36 via negative pressure and then forcibly discharged from the transfer chamber 36 towards its target chamber via positive pressure.

[0073] The transfer device 34 includes an enlarged end 37. In this arrangement, the enlarged end 37 is generally cylindrical and includes a drive unit in the form of teeth 38 located spaced apart around the enlarged end 37. The teeth 38 are configured to engage with the corresponding drive unit of System 1 such that the rotation of the corresponding drive unit of System 1 causes the transfer device 34 to rotate. The movement of the transfer device is controlled by a motor housed within the housing of System 1. The motor is a brushless DC motor, a stepper motor, or any type of electronically driven motor.

[0074] Referring next to Figures 8 and 9 of the attached drawings, the transfer device 34 includes a movable channel 39 defined by an internal passage within the enlarged end 37. The movable channel 39 is configured to move relative to the assay apparatus body 24 together with the transfer device 34. The transfer device 34 includes flow openings 40, 41 which are fluidically coupled to the movable channel 39. The flow openings 40, 41 are positioned so as to selectively align with openings 33 on the assay apparatus body 24 in order to selectively fluidize each chamber 25-30 to the movable channel 39 depending on the relative position of the transfer device 34 to the assay apparatus body 24.

[0075] One of the flow openings 40 is fluidically coupled to the transfer chamber 36 to allow fluid to flow into or out of the transfer chamber 36 when the piston 22 moves along at least a portion of the length of the transfer chamber 36 due to the positive or negative pressure generated in the transfer chamber 36 as a result of the movement of the piston 22.

[0076] The transfer device 34 includes a filtration arrangement 42 located within the enlarged end 37, such that the fluid flowing along the movable channel 39 passes through the filtration arrangement 42. In this arrangement, the filtration arrangement 42 comprises an array of filters, gaskets, and microbeads designed to separate large contaminants from cells contained in the sample and capture the cells within a “dissolution region”.

[0077] Referring to Figure 10 of the attached drawings, the filtration configuration 42 comprises at least one filter element. In this configuration, the filtration configuration 42 comprises a first filter element 43 with pores 2 μm to 30 μm in diameter, designed to filter out contaminants such as hair or dust. In this configuration, the filtration configuration 42 comprises a second filter element 44 superimposed on the first filter element 33. The second filter element 44 has pores between 0.1 μm and 5 μm in diameter, and the pore size is selected to be slightly smaller than the average size of target cells, so that target cells cannot pass through the second filter element 44.

[0078] In this configuration, the filtration configuration 42 includes gaskets 45-47 that seal around the filter elements 42 and 43. In this configuration, a larger gasket (approximately 200 μm thick) is provided between the first and second filter elements 43 and 44, forming a space for the dissolution region between the first and second filters.

[0079] In this configuration, the filtration configuration 42 comprises several beads B held between a first filter element 43 and a second filter 44. In some configurations, the beads B are microbeads with a diameter of approximately 100 microns. In some configurations, about half of the beads B are designed to have buoyancy so that they gather near the top of the filter configuration 42 during sonication, while the other half are designed to have no buoyancy and gather near the bottom of the filter configuration 42. Between the two types of beads, the majority of the dissolution area is filled with microbeads, which help to break down cell membranes during sonication.

[0080] Referring here to Figure 11 of the attached drawings, the transfer device 34 is positioned adjacent to the filtration arrangement 42 and includes an ultrasonic treatment chamber 48 that is fluidly coupled to the movable channel 39. In some arrangements, the ultrasonic treatment chamber 48 has a volume between 100 μl and 1000 μl. In some arrangements, the inlet to the ultrasonic treatment chamber 48 is positioned below the outlet of the ultrasonic treatment chamber 48 when the assay device 2 is upright, so that the liquid flows from lower to higher and any bubbles escape during the process.

[0081] The filtration configuration 42 is located within the sonication chamber, and the ultrasonic transducer 49 is located at one end of the sonication chamber 48. In some configurations, the filtration configuration 42 separates the inlet and outlet regions of the sonication chamber 48 by a distance of substantially half to one-quarter of the distance between the inlet and outlet of the sonication chamber 48.

[0082] The ultrasonic transducer 49 is electrically coupled to the controller 23 of system 1 when the assay apparatus 2 is inserted into system 1. The ultrasonic transducer 49 is configured to be controlled by the controller 23. The controller 23 comprises a processor and memory configured to control at least one process of the system, the memory storing executable instructions that, when executed by the processor, cause the processor to provide an output that performs at least one process. The memory of the controller 23 stores executable instructions that, when executed by the processor, cause the processor to control the ultrasonic transducer 49 and vibrate it at a selected frequency in order to lyse cells in the sonication chamber 48 and release nucleic acids (DNA / RNA) from the cells.

[0083] In some configurations, the ultrasonic transducer 49 is a compound containing, at least in part, lead, zirconium, and titanium. The compound of the ultrasonic transducer 49 is selected to give it properties for vibrating at frequencies of about 2.8 MHz to about 3.2 MHz. This frequency range is a preferred frequency range in which the ultrasonic transducer 49 generates ultrasound that dissolves or ruptures cells.

[0084] In some configurations, the ultrasonic transducer 49 comprises an upper first electrode and a lower second electrode located on the opposite side of the ultrasonic transducer 49. In some configurations, the first and second electrodes are made of silver, such as in the form of silver stamp paint. In some configurations, the capacitance between the first and second electrodes is 800 pF to 1300 pF.

[0085] In some configurations, the upper first electrode of the ultrasonic transducer 49 is covered with at least a portion of a glass coating. The glass coating minimizes or prevents contamination of the liquid in the ultrasonic chamber 48 by the material of the first electrode. The glass coating can also minimize or prevent erosion of the silver of the first electrode by cavitation bubbles collapse caused by ultrasound traveling through the liquid in the ultrasonic chamber 48 during system use, for example.

[0086] The first and second electrodes of the ultrasonic transducer 49 are electrically connected to the first and second electrical terminals of the controller 23, respectively.

[0087] In some configurations, the controller 23 includes an AC driver. The AC driver generates an AC drive signal of a predetermined frequency and outputs an AC drive signal to drive the ultrasonic transducer 49. The AC driver consists of a circuit incorporating electronic components connected to generate the AC drive signal from power received from the power supply. In some configurations, the AC driver includes an H-bridge circuit. In some configurations, the H-bridge circuit includes four MOSFETs connected to convert DC to AC at high frequencies (e.g., frequencies in the range of 2.8 MHz to 3.2 MHz).

[0088] In some configurations, the controller 23 includes an active power monitor. The active power monitor includes electronic circuitry that monitors the active power used by the ultrasonic transducer 49 when the ultrasonic transducer 49 is driven by an AC drive signal. The active power monitor provides a monitoring signal indicating the active power used by the ultrasonic transducer 49. In some configurations, the active power monitor includes a current sensor that senses the drive current of the AC drive signal driving the ultrasonic transducer 49 and provides a monitoring signal indicating the sensed drive current.

[0089] The processor in controller 23 controls the AC driver and receives monitoring signals from the active power monitor.

[0090] In some configurations, the controller 23 includes a frequency controller and is implemented in executable code stored in memory. This executable code, when executed by the processor, performs at least one function.

[0091] The memory of the controller 23 stores executable instructions that, when executed by the processor, control the ultrasonic transducer 49 to vibrate at multiple frequencies within a predetermined sweep frequency range, and to select the drive frequency of the ultrasonic transducer 49 between a first predetermined frequency and a second predetermined frequency for lysing cells in the sonication chamber 48.

[0092] In some arrangements, certain cells may require different frequencies due to their physical characteristics (size, shape, presence of a cell wall, etc.), so the frequency is determined by the type of cell being lysed.

[0093] There is an optimal frequency or frequency range for lysing cells within an ultrasonic chamber. The optimal frequency or frequency range depends on at least the following four parameters:

[0094] 1. Transducer manufacturing process In some configurations, the ultrasonic transducer 49 is equipped with a piezoelectric ceramic. The piezoelectric ceramic is manufactured by mixing compounds to create a ceramic base, and this mixing process may not be consistent throughout the manufacturing process. This lack of consistency can result in variations in the resonant frequency of the cured piezoelectric ceramic.

[0095] If the resonant frequency of the piezoelectric ceramic does not correspond to the required operating frequency, the cell lysis process will not be optimal. Even a slight deviation in the piezoelectric ceramic's resonant frequency will affect the lysis process, meaning the system will not function optimally.

[0096] 2. Transducer load During operation, changes in the load on the ultrasonic transducer 49 inhibit the overall displacement of the ultrasonic transducer 49's vibration. To achieve optimal vibration displacement of the ultrasonic transducer 49, the drive frequency needs to be adjusted so that the controller 23 can supply sufficient power for the maximum displacement.

[0097] The types of loads that may affect the efficiency of the ultrasonic transducer 49 may include the amount of liquid on the transducer (i.e., the amount of liquid in the ultrasonic processing chamber 48).

[0098] 3. Temperature The ultrasonic vibrations of the ultrasonic transducer 49 are partially attenuated by its assembly into the assay device 2. This attenuation of vibrations may cause a localized temperature rise on and around the ultrasonic transducer 49.

[0099] An increase in temperature affects the vibration of the ultrasonic transducer 49 due to changes in its molecular behavior. A rise in temperature means that more energy is imparted to the ceramic molecules, temporarily affecting their crystal structure. As the temperature decreases, the effect reverses, but modulation of the supply frequency is necessary to maintain optimal vibration.

[0100] Since an increase in temperature also reduces the viscosity of the solution in the sonication chamber 48, it may be necessary to change the drive frequency to optimize the lysis of cells in the sonication chamber 48.

[0101] 4. Distance to the power source The vibration frequency of the ultrasonic transducer 49 can vary depending on the wiring length between the ultrasonic transducer 49 and the oscillator driver. The frequency of the electronic circuit is inversely proportional to the distance between the ultrasonic transducer 49 and the controller 23.

[0102] While the distance parameter is primarily fixed in this configuration, it may change during the manufacturing process of System 1. Therefore, it is desirable to change the drive frequency of the ultrasonic transducer 49 to compensate for the fluctuations and optimize the efficiency of the system.

[0103] The ultrasonic transducer 49 can be modeled as an RLC circuit in an electronic circuit, as shown in Figure 12. The four parameters mentioned above change the resonant frequency range supplied to the transducer and can be modeled as changes in the inductance, capacitance, and / or resistance of the entire RLC circuit. As the circuit frequency rises to near the transducer's resonance point, the logarithmic impedance of the entire circuit drops to a minimum, then rises to a maximum, and settles within the median range.

[0104] Figure 13 is a general graph illustrating the change in overall impedance with increasing frequency in an RLC circuit. Figure 14 shows a piezoelectric transducer at a first predetermined frequency f s At the following frequencies, in the first capacitive region, the second predetermined frequency f p The above frequencies show how it acts as a capacitor in the second capacitive region. The piezoelectric transducer operates at the first and second predetermined frequencies f s ,f p Between these frequencies, it acts as an inductor in the inductive region. To maintain the transducer's optimal oscillation, i.e., maximum efficiency, the current flowing through the transducer must be kept at a frequency within the inductive region.

[0105] The memory of the controller 23 stores executable instructions that, when executed by the processor, cause the processor to maintain the vibration frequency of the ultrasonic transducer 49 within the induction range in order to maximize the lysis efficiency of cells in the sonication chamber 48.

[0106] The memory of the controller 23 stores executable instructions that, when executed by the processor, cause the processor to perform a sweep operation that drives the transducer at a frequency that the controller 23 progressively tracks over a predetermined sweep frequency range. In other words, the drive unit 2 drives the transducer at multiple different frequencies over a predetermined sweep frequency range. For example, frequencies that increment by a predetermined frequency from one end of the sweep frequency range to the other.

[0107] In some configurations, when the controller 23 performs a sweep, the controller 23 monitors the ADC (Analog-to-Digital Conversion) value of an analog-to-digital converter located within the controller 23 and coupled to the ultrasonic transducer 49. In some configurations, the ADC value is an ADC parameter proportional to the overall voltage of the ultrasonic transducer 49. In other configurations, the ADC value is an ADC parameter proportional to the current flowing through the ultrasonic transducer 49.

[0108] During the sweep operation, the controller 23 identifies the induction region of the transducer frequency. Once the controller 23 has identified the induction region, the controller 23 records the ADC value and optimizes the operation of the ultrasonic transducer 49 by setting the transducer's drive frequency to a frequency within the induction region (i.e., a first and second predetermined frequency f s ,f p The drive frequency locks within the inductive region. When the drive frequency is locked within the inductive region, the electromechanical coupling coefficient of the transducer is maximized, thereby maximizing the operation of the ultrasonic transducer 49.

[0109] In some configurations, the controller 23 determines the active power used by the ultrasonic transducer 49 by monitoring the current flowing through the transducer 49. Active power is the actual power or true power dissipated by the ultrasonic transducer 49.

[0110] The mechanical deformation of an ultrasonic (piezoelectric) transducer is linked to the amplitude of the AC voltage applied to it, and to ensure optimal system function and delivery, the ultrasonic transducer must always be supplied with maximum deformation. Pulse width modulation (PWM) of the AC voltage applied to the ultrasonic transducer keeps the mechanical amplitude of the vibration constant. In some configurations, the system actively adjusts the duty cycle of the AC voltage waveform to maximize the deformation of the ultrasonic transducer to ensure optimal system function and delivery.

[0111] One approach involves modifying the AC voltage applied to the ultrasonic transducer through the use of a digital-to-analog converter (DAC). While the energy transmitted to the ultrasonic probe decreases, the mechanical deformation also decreases, resulting in less deformation than the maximum possible. The effective voltage across the ultrasonic transducer remains the same as with voltage modulation, but the active power transmitted to the ultrasonic transducer degrades. In fact, this can be expressed by the following equation: The active power displayed on the ultrasonic transducer is as follows:

[0112]

number

[0113] Here

[0114]

number

[0115] I rms This is the root mean square current. V rms This is the mean square voltage.

[0116] When considering the first harmonic, pulse width modulation changes the duration of the voltage supplied to the ultrasonic transducer, thereby controlling Irms, so Irms is a function of the actual voltage amplitude applied to the ultrasonic transducer.

[0117] In this configuration, the memory of the controller 23 stores instructions that, when executed by the processor of the controller 23, cause the processor to perform the following actions:

[0118] A. The AC driver of the controller 23 is controlled to cause the ultrasonic transducer 49 to output an AC drive signal at a predetermined sweep frequency.

[0119] B. Based on the monitoring signal, calculate the active power used by the ultrasonic transducer 49.

[0120] C. Control the AC driver to modulate the AC drive signal and maximize the active power used by the ultrasonic transducer 49. D. Record the maximum active power used by the ultrasonic transducer 49 and the sweep frequency of the AC drive signal and save them to memory.

[0121] E. After a predetermined number of iterations, steps A to D are repeated for a predetermined number of iterations, with the sweep frequency incremented in each iteration, so that the sweep frequency increases from the sweep start frequency to the sweep end frequency.

[0122] F. From the records stored in memory, identify the optimal frequency of the AC drive signal, which is the sweep frequency of the AC drive signal at which the maximum active power is used by the ultrasonic transducer 49, and G. Control the AC driver to output an AC drive signal to the ultrasonic transducer 49 at the optimal frequency.

[0123] In some configurations, the starting sweep frequency is 2800 kHz and the ending sweep frequency is 3200 kHz. In other configurations, the starting and ending sweep frequencies are the lower and upper limits of the frequency range between 2800 kHz and 3200 kHz.

[0124] In some configurations, the processor controls the AC driver to output an AC drive signal to the ultrasonic transducer 49 at a frequency shifted by only 1-10% of the optimal frequency. In these configurations, the frequency shift is used to extend the service life of the ultrasonic transducer 49 by minimizing the potential damage that would occur to the ultrasonic transducer 49 if it were continuously driven at the optimal drive frequency that produces the maximum displacement.

[0125] In some configurations, the AC driver modulates the AC drive signal by pulse width modulation to maximize the active power used by the ultrasonic transducer 49.

[0126] In some configurations, the processor 40 controls the AC driver to alternately output an AC drive signal to the ultrasonic transducer 49 at an optimal frequency for a first predetermined time, and then to a second predetermined time, by not outputting an AC drive signal to the ultrasonic transducer 49. This alternating activation and deactivation of the ultrasonic transducer 49 has been found to optimize the process of lysing cells in the sample within the sonication chamber 48.

[0127] In some configurations, the controller 23 operates in recursive mode to ensure optimal operation of the ultrasonic transducer 49. When the controller 23 operates in recursive mode, it periodically performs a sweep of the frequencies in steps A to D during the operation of the system.

[0128] In some configurations, the AC driver of controller 23 is configured to alternately output AC drive signals and not output AC drive signals according to the operating mode. The timing of the 12 operating modes in some configurations is shown in Table 1 below.

[0129] [Table 3]

[0130] In some configurations, the memory of the controller 23 stores executable instructions that, when executed by the processor, cause the processor to perform a sweep operation to identify the inductive region each time vibration is started or restarted. In these configurations, the memory of the controller 23 stores executable instructions that, when executed by the processor, cause the processor to lock the drive frequency at a new frequency within the inductive region each time vibration is started, thereby compensating for parameter changes that affect the operating efficiency of the ultrasonic transducer 49.

[0131] In some configurations, the controller 23 operates in recursive mode to ensure optimal operation of the ultrasonic transducer 49. When the controller 23 operates in recursive mode, it periodically sweeps the frequency during system operation and monitors the ADC value to determine whether the ADC value is above a predetermined threshold indicating optimal oscillation for the ultrasonic transducer 49.

[0132] In some configurations, the controller 23 performs a sweep operation during the cell lysis process if the controller 23 can identify a possible and better frequency for the ultrasonic transducer 49 that maximizes the displacement of the ultrasonic transducer 49. If the controller 23 identifies a better frequency, the controller 23 locks the drive frequency to the newly identified better frequency in order to maintain optimal operation of the ultrasonic transducer 49.

[0133] Figure 15 is a flowchart of the operation of controller 23 in some configurations.

[0134] Referring next to Figures 16 and 17 of the attached drawings, the lid 14 of the assay apparatus 2 includes a generally planar cover 50 configured to close at least the open end of the sample chamber 25 of the assay apparatus body 24. The lid 14 includes side walls 51 extending around the periphery of the cover 50. In this configuration, the air intake opening 52 is provided in one of the side walls 51.

[0135] In this configuration, the lid 14 is provided with a pivot mounting arrangement 53 for pivotally attaching the lid 14 to the assay apparatus body 24. In another configuration, the lid 14 is configured with different movable mounting arrangements for movably attaching the lid 14 to the assay apparatus body 24.

[0136] The lid 14 includes a gas-permeable membrane 54 that overlaps the lid member 50 around the ends of the side walls 51. The gas-permeable membrane 54 provides a substantially airtight seal around the side walls 51 and around the central opening 15, preventing cross-contamination or accidental spillage. In some configurations, the gas-permeable membrane 54 is made of Gore-Tex® material.

[0137] During use, the air intake opening 52 allows air to flow into the lid 14, and that air to pass through the gas permeable membrane 54 and flow into at least the sample chamber 25 inside the assay apparatus body 24.

[0138] In an alternative configuration, the gas-permeable membrane 54 may be replaced by another unidirectional gas flow member, such as a valve.

[0139] Referring here to Figures 18-20 of the attached drawings, the PCR apparatus 16 of the assay apparatus 2 is equipped with a fin 55, which is coupled to the assay apparatus body 24 so as to protrude outward from the assay apparatus body 24. The fin 55 is equipped with an enlarged mounting member 56 configured to be connected to the assay apparatus body 24. The mounting member 56 is provided with a first opening 57 and a second opening 58 that extend through the fin 55 to allow fluid communication with a PCR chamber 59 defined within the fin 55. In this configuration, the fin 55 further comprises a plurality of internal chambers 60 in a central portion 61 that partially surrounds the PCR chamber 59.

[0140] The fin 55 is roughly rectangular with angled ends 62, 63 converging at point 64. In use, the sample passes through both reagent chambers of the assay apparatus 2 and is then pushed into the PCR fin 55, which contains the PCR chamber 59.

[0141] In some configurations, the reagents selected for the PCR step are chosen to facilitate the extreme rRT-PCR step while also allowing for fluorescence-based temperature monitoring. In some configurations, the reagent formulation includes 5 μM of each forward and reverse primer (a total of 6 primers, 2 sets for detecting SARS-CoV-2 and 1 set to be used as a control if the PCR reaction is successful), IX LCGreen+ dye, 0.2 μM of each deoxynucleoside triphosphate (dNTP): dATP, dTTP, dGTP, dCTP, 50 mM Tris, 1.65 μM KlenTaq, 25 ng / μL BSA, 1.25 U / μL Malone Murine leukemia virus reverse transcriptase (MMLV), 7.4 mM MgCl2, and sulfotamine B.

[0142] Referring now to Figures 21 and 22 of the attached drawings, the fins 55 of the PCR device 16 are configured such that at least a portion of them are received inside the heating device 17.

[0143] In this configuration, the heating device 17 comprises two roughly circular planar discs 65, 66 that are spaced apart from each other and rotatably mounted on a pivot member 67. The heating recess 68 is defined by a portion of the space between the discs 65, 66.

[0144] In this configuration, one of the disks 65 is a movable support element that holds the first heating element 69 and the second heating element 70, as shown in Figure 23. The first and second heating elements 69 and 70 are arranged on both sides of the disk 65 with space between them.

[0145] The heating device 17 further includes a motor configured to rotate the disc 65 around a pivot member 67 such that the disc 65 operates between a first position in which the first heating element 69 is positioned closer to the heating recess 68 than the second heating element 70, and a second position in which the second heating element 70 is positioned closer to the heating recess 68 than the first heating element 69. The motor is electrically coupled to a controller 23, which controls the motor to periodically move the disc 65 between the first and second positions.

[0146] In some configurations, the heating device 17 includes a temperature sensor configured to sense the temperature of the liquid in the PCR device located within the heating recess 68, and the system is configured to control the operation of the first and second heating elements in response to the sensed temperature.

[0147] Next, referring to Figure 24 of the attached drawings, System 1 includes an infectious disease detection arrangement in the form of a fluorescence detection arrangement 70, which comprises a generally planar support member 71 having an opening 72 from which a pivot member 67 extends. The fluorescence detection arrangement comprises a first triangular portion 73, a second triangular portion 74, and a recessed portion 75. The planar body 71 and the triangular portions 73 and 74 are positioned in the space between the disks 65 and 66 of the heating device.

[0148] The recessed portion 75 is shaped to receive the pointed end of the fin 55 of the PCR device 16.

[0149] The detection device 70 comprises a plurality of light emitters 76 along one edge of the recessed portion 75 and a plurality of photoreceptors 77 along the other edge of the recessed portion 75. In this arrangement, there are four light emitters in the form of four LEDs, each configured to transmit light of a different wavelength, and four photoreceptors 77, each configured to detect light of a different wavelength. However, in an alternative arrangement, a different number of light emitters and photoreceptors are present.

[0150] In some configurations, the detection device 70 is configured to detect fluorescence emitted from LCGreen+ and sulfofodamine B dye in order to monitor PCR, melting curves, and temperature changes.

[0151] In some configurations, the detection device is a SARS-CoV-2 virus detection device, which detects the presence of the SARS-CoV-2 virus that causes COVID-19 disease.

[0152] Results report In some configurations, System 1 includes a display, such as an LCD monitor, on the outside of the enclosure 3. After information from the system is processed by the controller 23, the test results are displayed on the display. There are four possible test results: positive, negative, indeterminate, or invalid. For COVID-19 tests, the criteria for the four results are shown in Table 2 below.

[0153] [Table 4]

[0154] SARS-CoV-2 example Next, we will describe the operation of some configurations of the SARS-CoV-2 assay system.

[0155] In assay device 2, the first chamber is a sample chamber into which the user adds the target sample to be tested. In some configurations, the target sample is a saliva sample or a sputum sample. In other configurations, the target sample is collected from the user by a nasopharyngeal swab or an oropharyngeal swab. In yet another configuration, the target sample is a blood sample.

[0156] In some configurations, the volume of the target sample is between 1 ml and 5 ml. The sample taken from the patient is placed in elution buffer before being added to the sample chamber. In some configurations, the elution buffer contains 1 M imidazole solution, 1 M Tris, 0.5 M EDTA, Milli-Q, or deionized water.

[0157] The next chamber is the washing chamber. In some configurations, the washing chamber contains an excess volume (3 ml to 5 ml) of the elution buffer described above. This washing buffer is used to wash the sample and remove potential contaminants.

[0158] The next chamber is the solubilant chamber. In some configurations, the solubilant chamber contains a mixture of chemicals to assist the cell lysis step of the assay. In some configurations, the solubilant consists of formulations and includes, but is not limited to, the following three formulations: Solvent formulation #1: 10mM Tris 0.25% Igepal CA-630 150 mM NaCl Solvent formulation #2: 10 mM Tris-HCl 10 mM NaCl 10mM EDTA 0.5% Triton-X100 Solvent formulation #3: 0.1M LiCl 0.1M Tris-HCl 1% SDS 10mm EDTA The next chamber is the liquid reagent mixing chamber. The sample is sonicated, and once cell lysis occurs, the released nucleic acids are pushed into the liquid reagent mixing chamber by pressure from the plunger column. The liquid reagent chamber contains the liquid-stabilizing components of the rRT-PCR reagent mixture. Examples of components retained in this chamber, in some configurations, are Tris, IX LCGreen Dye, free nucleotides, MgCl2, or sulforodamine B.

[0159] The next chamber is a lyophilized reagent mixing chamber. This chamber contains lyophilized or lyophilized forms of reagents that cannot be stored for long periods in liquid or hydrated state, such as proteins. Examples of components that would be lyophilized for long-term storage in an assay instrument include, in some configurations, primers, polymerases, reverse transcriptases, or bovine serum albumin (BSA).

[0160] The next chamber is the PCR chamber, which is located outside the main section of the pod within the PCR fin. This chamber is where the final mixed PCR solution (containing nucleic acids released from the initial sample and all PCR reagents) is delivered before the rRT-PCR step.

[0161] The final chamber is the waste chamber. This chamber holds all components that are discarded throughout the assay apparatus cycle. For example, when the washing solution is pushed out of the sonication chamber, this solution is sent directly to the waste chamber as it exits the sonication chamber. The volume of this chamber must be at least equal to the sum of the total volume of all liquids in the pod plus the volume of the added sample.

[0162] PCR method In some configurations, rRT-PCR is performed to rapidly detect and confirm the presence of SARS-CoV-2 in a sample. To control the heating and cooling processes necessary for the RT-PCR reaction to occur, some configurations use a heating device 17 as a thermal cycler with a dual heating element that provides the required temperature cycles.

[0163] The disks 65 and 66 of the heating device 17 rapidly rotate during extreme rRT-PCR cycling, applying different heat levels to heat the PCR chamber to the desired temperature. Heating elements 69a and 69b are located on the opposite side of the disks, each occupying an area of ​​1 / 4 of the disk's surface area. Each heating element 69a and 69b is programmed to reach a predetermined temperature.

[0164] The first heating element 69a is initially heated to 45°C, paused for the reverse transcriptase step, and then heated to its PCR temperature of 55°C. The second heating element 69b is heated to 95°C and is used only in the PCR step. The other two sections of disk 65 serve as insulating regions between heating elements 69a and 69b.

[0165] In some configurations, the thermal cycling is performed as follows: The temperature of the first heating element 69a rises to 45°C while the PCR chamber is exposed to the insulating section of the disk. Once the first heating element 69a reaches 45°C, the disk 65 rotates to expose the PCR chamber to the second heating element 69b for 2 seconds, allowing the reverse transcriptase step to occur. Immediately thereafter, the first heating element is heated to 55°C to initiate the PCR step.

[0166] In some configurations, disk 65 is started by rapidly alternating exposure to the first and second heating elements of the PCR chamber for approximately 30–35 cycles of heating and cooling. After each rotation of disk 65, the temperature of the liquid in the PCR chamber is monitored using passive fluorescence detection of sulforodamine B dye.

[0167] The second heating element 69b is positioned close to the PCR chamber, and when the temperature of the liquid in the PCR chamber reaches 95°C, the disk 65 is triggered to rotate, moving the first heating element 69a adjacent to the PCR chamber. Then, when the chamber temperature drops to 55°C, the disk 65 rotates back to the second heating element 69b. This completes one cycle.

[0168] After the final PCR cycle, the first heating element 69a rotates to a position close to the PCR chamber and begins heating at a rate of 8°C / second to a temperature between 90°C and 100°C, allowing for melt analysis to be performed to confirm the presence of a specific PCR product.

[0169] System 1 can provide test results within 10 minutes, and in some configurations, it can be done in as little as 5 minutes. This is significantly faster than conventional PCR testing and opens up the possibility of rapid testing not only in homes, shops, and entertainment venues, but also in transportation hubs such as airports and bus or train terminals.

[0170] Some configurations of System 1 are highly portable and can be easily carried to wherever testing is needed. Due to the system's efficient operation, some configurations can be battery-powered, allowing the system to provide testing in virtually any location.

[0171] The above outlines some features of embodiments so that those skilled in the art may better understand the various aspects of this disclosure. Those skilled in the art should understand that this disclosure can be readily used as a basis for designing or modifying other processes and structures to accomplish the same objectives and / or achieve the same advantages of the various embodiments introduced herein. Furthermore, those skilled in the art should recognize that such equivalent structures will not deviate from the spirit and scope of this disclosure, and that various changes, substitutions, and modifications can be made to this disclosure without departing from the spirit and scope of this disclosure.

[0172] While the subject matter has been described using language specific to structural features or methodological actions, it should be understood that the subject matter of the attached claims is not necessarily limited to the specific features or actions described above. Rather, the specific features and actions described above are disclosed as exemplary forms of implementing at least a part of the claims.

[0173] This specification provides various operations of embodiments. The order in which some or all operations are described should not be construed to mean that these operations are necessarily order-dependent. It should be understood that alternative orders are of interest in this description. Furthermore, it should be understood that not all operations are necessarily present in every embodiment provided herein. Also, it should be understood that not all operations are necessary in some embodiments.

[0174] Furthermore, in this specification, “exemplary” means to be used as an example, instance, illustration, etc., and is not necessarily advantageous. “Or” as used in this application is intended to mean inclusive “or,” not exclusive “or.” Furthermore, “a” and “an” as used in this application and the attached claims are generally interpreted as meaning “one or more,” unless otherwise specified or it is clear from the context that they are directed to the singular form. Furthermore, “including,” “having,” “possessing,” “together,” or variations thereof, are intended to be inclusive in the same manner as the term “including.” Also, unless specifically stated, “first,” “second,” etc., are not intended to suggest temporal, spatial, or sequential aspects. Rather, such terms are used merely as identifiers, names, etc., for features, elements, items, etc. For example, the first element and the second element generally correspond to element A and element B, or two different elements or two identical elements, or the same element.

[0175] Furthermore, although this disclosure has been shown and described in relation to one or more embodiments, equivalent changes and modifications will be made to others skilled in the art based on a reading and understanding of this specification and the accompanying drawings. This disclosure includes all such changes and modifications and is limited only to the scope of the following claims. In particular, with respect to the various functions performed by the features described above (e.g., elements, resources, etc.), the terms used to describe such features are intended to correspond to any feature (e.g., functionally equivalent) that performs a particular function of the described feature, even if it is not structurally equivalent to the disclosed structure, unless otherwise indicated. In addition, although certain features of this disclosure may have been disclosed in relation to only one of several embodiments, such features may be combined with one or more other features of other embodiments as desired and advantageous for any or particular application.

[0176] The subject matter and functional operating embodiments described herein can be implemented in digital electronic circuits, computer software, firmware, or hardware, or in one or more combinations thereof, including the structures disclosed herein and their structural equivalents.

[0177] Features of some embodiments are implemented using one or more modules of computer program instructions encoded on a computer-readable medium to be executed by a data processing device or controller, or to control its operation. The computer-readable medium can be a manufactured product such as a hard drive in a computer system or embedded system. The computer-readable medium can be acquired separately, for example, by distribution of one or more modules of computer program instructions via a wired or wireless network, and subsequently encoded with one or more modules of computer program instructions. The computer-readable medium can be a machine-readable storage device, a machine-readable storage board, a storage device, or a combination of one or more of these.

[0178] The terms "computing device" and "data processing device" encompass all devices, machines, and equipment for processing data, including, for example, programmable processors, computers, or multiple processors or computers. In addition to hardware, a device may include code that constitutes the execution environment of the computer program, such as processor firmware, a protocol stack, a database management system, an operating system, a runtime environment, or one or more of these. Furthermore, such a device may employ various different computing models, such as web services, distributed computing, and grid computing infrastructure.

[0179] The processes and logical flows described herein can be executed by one or more programmable processors running one or more computer programs, by acting on input data and producing outputs.

[0180] As used herein, in some embodiments, the term "module" includes memory and / or a processor configured to control at least one process of a system or circuit structure. Memory stores executable instructions that, when executed by the processor, cause the processor to provide output for executing at least one process. Embodiments of memory include non-temporary computer-readable media.

[0181] Processors suitable for executing computer programs include, as an example, general-purpose and special-purpose microprocessors, as well as any one or more processors in any type of digital computer. Generally, a processor will receive instructions and data from read-only memory, random-access memory, or both. Essential elements of a computer are a processor for executing instructions and one or more memory devices for storing instructions and data. Generally, a computer includes one or more mass storage devices for storing data, such as magnetic disks, magneto-optical disks, or optical disks, or is operablely coupled to them to receive data from them, transfer data to them, or both. However, a computer is not required to have such devices. Devices suitable for storing computer program instructions and data include all forms of non-volatile memory, media, and memory devices, including, as examples, semiconductor memory devices such as EPROM (Erasable Programmable Read-Only Memory) and EEPROM (Electrically Erasable Programmable Read-Only Memory), and magnetic disks such as internal hard disks or removable disks, magneto-optical disks, and flash memory devices such as CD-ROMs and DVD-ROMs.

[0182] To provide user interaction, some embodiments are implemented in a computer equipped with a display device for displaying information to the user, such as a CRT (cathode ray tube) or LCD (liquid crystal display) monitor, and a keyboard and pointing device that allows the user to provide input to the computer, such as a mouse or trackball. Other types of devices may also be used to provide user interaction. For example, the feedback provided to the user can be any form of sensory feedback, such as visual feedback, auditory feedback, or haptic feedback, and the input from the user can be received in any form, such as acoustic, voice, or haptic input.

[0183] In this specification, "equipped" means "to include or constitute," and "equipped" means "to include or constitute."

[0184] The features disclosed in the preceding description, the following claims, or the accompanying drawings may be expressed as appropriate in their specific forms, or in terms of means for performing the disclosed functions, or methods or processes for achieving the disclosed results, and may be used individually or in any combination of those features to realize the invention in its various forms.

Claims

1. Infectious disease testing system, An ultrasonic processing chamber configured to accept biological samples for infectious disease testing, An ultrasonic transducer configured to output ultrasound in the frequency range of 2800 kHz to 3200 kHz in order to lyse cells from a biological sample in the sonication chamber, It is a controller, An AC driver configured to generate an AC drive signal at a predetermined frequency within a frequency range of 2800 kHz to 3200 kHz, and to output the AC drive signal for driving the ultrasonic transducer, An active power monitor configured to monitor the active power used by the ultrasonic transducer when the ultrasonic transducer is driven by an AC drive signal, the active power monitor configured to provide a monitoring signal indicating the active power used by the ultrasonic transducer, A processor configured to control the AC driver and receive the monitoring signal from the active power monitor, A memory for storing instructions, which, when executed by the processor, the processor, A. Control the AC driver to output an AC drive signal to the ultrasonic transducer at a predetermined sweep frequency. B. Based on the monitoring signal, calculate the active power used by the ultrasonic transducer. C. Control the AC driver to modulate the AC drive signal and maximize the active power used by the ultrasonic transducer. D. The maximum active power used by the ultrasonic transducer and the sweep frequency of the AC drive signal are recorded and stored in the memory. E. After a predetermined number of repetitions, steps A to D are repeated for a predetermined number of repetitions, with the sweep frequency increasing with each repetition so that the sweep frequency increases from the sweep start frequency to the sweep end frequency. F. From the records stored in the memory, identify the optimal frequency of the AC drive signal, which is the sweep frequency of the AC drive signal in which the maximum active power is used by the ultrasonic transducer, and G. A controller comprising a memory that controls the AC driver and outputs the AC drive signal to the ultrasonic transducer at the predetermined optimal frequency, The system further, A polymerase chain reaction ("PCR") apparatus configured to receive and amplify DNA from lysed cells of a biological sample, and An infectious disease testing system characterized by comprising an infectious disease detection device configured to detect the presence of an infectious disease in amplified DNA and to provide an output indicating whether or not the presence of an infectious disease in the amplified DNA has been detected.

2. The aforementioned active power monitor, The infectious disease testing system according to claim 1, comprising a current sensor configured to sense the drive current of the AC drive signal that drives the ultrasonic transducer, wherein the active power monitor is configured to provide a monitoring signal indicating the sensed drive current.

3. The infectious disease testing system according to claim 1 or 2, characterized in that when the memory is executed by the processor, the system stores an instruction that causes the processor to repeat steps A to D while increasing the sweep frequency from a sweep start frequency of 2800 kHz to a sweep end frequency of 3200 kHz.

4. When the memory is executed by the processor, the processor will The infectious disease testing system according to any one of claims 1 to 3, characterized in that step G stores a command to control the AC driver to output the AC drive signal at a frequency shifted by 1 to 10% from the optimal frequency to the ultrasonic transducer.

5. The infectious disease testing system according to any one of claims 1 to 4, wherein the AC driver is configured to pulse-width modulate the AC drive signal in order to maximize the active power used by the ultrasonic transducer.

6. When the memory is executed by the processor, the processor will The infectious disease testing system according to any one of claims 1 to 5, characterized in that it records commands to alternately control the AC driver, outputting an AC drive signal of the optimal frequency to the ultrasonic transducer for a first predetermined time, and not outputting the AC drive signal to the ultrasonic transducer for a second predetermined time.

7. When the memory is executed by the processor, the processor will The infectious disease testing system according to claim 6, characterized in that it stores commands to alternately output the AC drive signal and to not output the AC drive signal according to an operation mode selected from the following. Table 1

8. The aforementioned infectious disease testing system, A heating device, A heating recess for receiving a part of the PCR device, Movable support elements, A first heating element supported by the aforementioned support element, A second heating element, which is transported by the support element at a position spaced apart from the first heating element, wherein the support element is movable between a first position in which the first heating element is positioned closer to the heating recess than the second heating element, and a second position in which the second heating element is positioned closer to the heating recess than the first heating element, and An infectious disease testing system according to any one of claims 1 to 7, further comprising a heating device including a motor configured to periodically move the support element between a first position and a second position.

9. The aforementioned heating device The infectious disease testing system according to claim 8, further comprising a temperature sensor configured to sense the temperature of the liquid in the PCR device disposed within the heating recess, wherein the controller is configured to control the movement of the first and second heating elements in response to the sensed temperature.

10. The infectious disease testing system according to claim 8 or 9, characterized in that the controller is configured to control a first heating element to substantially heat the liquid in the PCR apparatus to 45°C during the reverse transcriptase step.

11. During the PCR process, the controller The first heating element is controlled so as to substantially heat the liquid in the PCR apparatus to 55°C. The second heating element is controlled to heat the liquid in the PCR apparatus to substantially 95°C. The infectious disease testing system according to claim 9, characterized in that the first and second heating elements are configured to circulate the support element between the first and second positions such that the temperature of the liquid in the PCR apparatus circulates substantially between 55°C and substantially 95°C.

12. The aforementioned system The infectious disease testing system according to any one of claims 1 to 11, further comprising a movable channel to selectively provide a fluid flow path between the sample chamber, the sonication chamber, or the PCR chamber, so that at least a portion of the sample can be continuously transferred between the sample chamber, the sonication chamber, and the PCR chamber.

13. The aforementioned system The infectious disease testing system according to claim 12, further comprising a filtration arrangement configured to filter fluid flowing out of the movable channel, the filtration arrangement further comprising a first filter element having holes with a diameter of 2 μm or more and 30 μm or less.

14. The infectious disease testing system according to claim 13, characterized in that the filtration arrangement comprises a second filter element superimposed on the first filter element, and the pores of the second filter element are 0.1 μm to 5 μm in diameter.

15. The infectious disease testing system according to any one of claims 1 to 14, characterized in that the system is a COVID-19 infection testing system, and the infectious disease detection device is a SARS-CoV-2 virus detection device configured to detect the presence of the SARS-CoV-2 virus that causes COVID-19 infection in amplified DNA and to provide an output indicating whether or not the SARS-CoV-2 virus detection device detects the presence of COVID-19 disease in the amplified DNA.

16. An infectious disease testing method, wherein the method is An ultrasonic processing chamber is provided, which includes an ultrasonic transducer that outputs ultrasound in the frequency range of 2800 kHz to 3200 kHz in order to dissolve cells obtained from a biological sample within the ultrasonic processing chamber, and a biological sample to be tested for infectious diseases is placed in the ultrasonic processing chamber. The AC driver generates an AC drive signal at a predetermined frequency within the frequency range of 2800 kHz to 3200 kHz, and outputs the AC drive signal to an ultrasonic transducer to drive the ultrasonic transducer. The active power monitor monitors the active power used by the ultrasonic transducer when the ultrasonic transducer is driven by the AC drive signal, and the active power monitor provides a monitoring signal indicating the active power used by the ultrasonic transducer. The processor includes receiving an active signal from the active power monitor, and further the method is A. The processor controls the AC driver to output the AC drive signal to the ultrasonic transducer at a predetermined sweep frequency, B. The processor calculates the active power used in the ultrasonic transducer based on the monitoring signal, C. The processor modulates the AC drive signal and controls the AC driver to maximize the active power used in the ultrasonic transducer, D. The processor stores in memory the record of the maximum active power used by the ultrasonic transducer and the sweep frequency of the AC drive signal, E. After a predetermined number of repetitions, steps A to D are repeated a predetermined number of times with the sweep frequency incremented in each repetition, such that the sweep frequency increases from the sweep start frequency to the sweep end frequency. F. The processor identifies, from the records stored in the memory, the optimal frequency of the AC drive signal, which is the sweep frequency of the AC drive signal in which the maximum active power is used by the ultrasonic transducer, and G. The processor controls the AC driver to output the AC drive signal to the ultrasonic transducer at the predetermined optimal frequency, Furthermore, the above method, The polymerase chain reaction (PCR) device receives DNA from lysed cells in a biological sample and amplifies it. In an infectious disease detection device, the presence of pathogens causing infectious diseases in the amplified DNA is detected, An infectious disease testing method comprising providing an output indicating whether or not the infectious disease detection device has detected the presence of an infectious disease-causing pathogen in the amplified DNA.

17. The aforementioned method, The current sensor detects the drive current of the AC drive signal that drives the ultrasonic transducer, The infectious disease testing method according to claim 16, further comprising providing a monitoring signal indicating the sensed drive current using the active power monitor.

18. The aforementioned method, The infectious disease testing method according to claim 16 or 17, further comprising modulating the AC drive signal to a pulse width using the AC driver to maximize the active power used in the ultrasonic transducer.

19. The aforementioned method, The infectious disease testing method according to any one of claims 16 to 18, further comprising alternately controlling an AC driver to output the AC drive signal of the optimal frequency to the ultrasonic transducer for a first predetermined time, and not outputting the AC drive signal to the ultrasonic transducer for a second predetermined time.

20. The aforementioned method, The infectious disease testing method according to claim 19, further comprising controlling the AC driver according to an operating mode selected from the following, thereby alternately controlling the output and non-output of the AC drive signal. Table 2

21. The above method is a COVID-19 infection testing method, and the above method is In a SARS-CoV-2 virus detection device, the presence of the SARS-CoV-2 virus, which causes COVID-19 infection, is detected in the amplified DNA. An infectious disease testing method according to any one of claims 16 to 20, characterized in that it includes providing an output indicating whether or not the SARS-CoV-2 virus detection device has detected the presence of COVID-19 disease in the amplified DNA.

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