Method and apparatus for controlling fluid volumes to achieve separation and PCR amplification

Abstract

A control device for a fluid volume comprising a motor, a camshaft connected to the motor at an axis of rotation of the camshaft, at least one cam arranged on a circumference of the camshaft, a pin frame, and at least one pin arranged within the pin frame and operatively associated with the at least one cam, wherein rotation of the camshaft by the motor causes the at least one cam to contact the at least one pin and drives the at least one pin in a first direction.

Description

(Cross-reference to related applications)

[0001] This application claims the benefit of priority under PCT Article 8 of U.S. Provisional Patent Application No. 63 / 093,640, filed October 19, 2020, and entitled "Point of Collection qPCR System." This application is also related to PCT applications entitled "Fluidic Detection and Control Algorithm for PCR Analysis," "Disposable Cartridge for Reagent Storage and Methods Using Same," and "Apparatuses with Fluidic Channel Geometry for Sample to Answer PCR Analysis and Methods of Using Same," all filed concurrently on October 19, 2021, and listing the same applicant, Formulatrix, Inc., and U.S. Design Application No. 29 / 812,034, entitled "Fluidic Channel Geometry of a Chip." The contents of all of the above applications are incorporated by reference in their entireties as if fully set forth herein. [Technical Field]

[0002] The present invention, in some embodiments thereof, relates to fluid flow, and more particularly, but not exclusively, to devices and methods for rapidly and efficiently moving small volumes of fluid. [Background technology]

[0003] Most current approaches to moving microliter-scale liquids involve mechanically complex approaches. Consider a syringe with a piston sealed against a cylinder. In most systems, this is a direct method of applying pressure or vacuum, but considering the sealing force (an O-ring or sealing interface sliding against the cylinder), driving the piston up and down is typically achieved by a motor rotating a ball screw to drive the relative motion of the piston with respect to the cylinder.

[0004] Peristaltic pumps are another simpler method, but they add or remove discrete volumes of gas or liquid, which may not be desirable in some applications.

[0005] Another method is the centrifugal device, so-called "cd-microfluidics," which uses different rotational speeds and interfacial characteristics to achieve liquid movement. See ufluidix.com / circle / whats-a-discman-and-how-is-it-a-medical-diagnostic-device-cd-microfluidics / . While using centrifugal devices may be convenient for some workflows, certain processes, such as real-time quantitative polymerase chain reaction ("qPCR"), cannot currently utilize this mechanism effectively. Summary of the Invention

[0006] According to an aspect of some embodiments of the present invention, there is provided an apparatus for controlling a fluid volume comprising: a motor; a camshaft coupled to the motor at an axis of rotation of the camshaft; at least one cam disposed on a circumference of the camshaft; a pin frame; and at least one pin disposed within the pin frame and operatively associated with the at least one cam, wherein rotation of the camshaft by the motor causes the at least one cam to contact the at least one pin and drive the at least one pin in a first direction.

[0007] In one embodiment of the present invention, the camshaft includes a plurality of cams and a plurality of pins, each of the plurality of cams corresponding to one of the plurality of pins.

[0008] In one embodiment of the present invention, multiple cams are arranged around the circumference of the camshaft such that rotation of the camshaft about the axis of rotation utilizes each of the multiple cams to drive a corresponding pin, thereby resulting in actuation of the multiple pins in a desired timing and sequence.

[0009] In one embodiment of the invention, the device further comprises a cartridge including a flexible, elastic membrane, the membrane being disposed between the cartridge and the at least one pin.

[0010] In one embodiment of the invention, the cartridge includes at least one well formed therein that corresponds to at least one pin.

[0011] In one embodiment of the invention, at least one pin is spring-loaded by a flexible, elastic membrane.

[0012] In one embodiment of the present invention, the at least one pin is provided by a flexible and elastic membrane so as to be movable in a second direction opposite to the first direction.

[0013] In one embodiment of the present invention, the pin frame comprises at least one slot through which at least one pin passes.

[0014] In one embodiment of the invention, a slot in the pin frame positions at least one pin over a well in the cartridge, with the at least one pin positioned between the cartridge and the at least one cam.

[0015] According to another aspect of some embodiments of the present invention, there is provided a system for performing real-time qPCR analysis, the system comprising: a fluid volume control device as described in claim 1; a cartridge comprising a membrane and at least one well; and a chip, wherein at least one well of the cartridge is disposed between the membrane and the chip.

[0016] In one embodiment of the present invention, the camshaft includes a plurality of cams and a plurality of pins, each of the plurality of cams corresponding to one of the plurality of pins.

[0017] In one embodiment of the present invention, multiple cams are arranged around the circumference of the camshaft such that rotation of the camshaft about the axis of rotation utilizes each of the multiple cams to drive a corresponding pin, thereby resulting in actuation of the multiple pins in a desired timing and sequence.

[0018] In one embodiment of the present invention, at least one cam drives at least one pin into at least one well.

[0019] In one embodiment of the invention, the membrane is elastic.

[0020] In one embodiment of the invention, a membrane is disposed between at least one pin and at least one well, such that the membrane forms a fluid seal with the well when driven into the well by the at least one pin.

[0021] In one embodiment of the invention, at least one pin is provided with movement in a second direction opposite the first direction by a flexible, elastic membrane.

[0022] According to another aspect of some embodiments of the present invention, there is provided a method of controlling fluid volume in a real-time qPCR system, the method including rotating a camshaft about an axis of rotation with a motor, contacting at least one pin with a cam located on the camshaft, driving the at least one pin in a first direction with the cam, depressing a membrane in a well of a cartridge with the at least one pin, and expelling fluid in the well with the at least one pin and the membrane.

[0023] In one embodiment of the invention, the method further includes maintaining rotation by driving at least one additional pin with the at least one cam and using the additional pin and membrane to push additional fluid in additional wells.

[0024] In one embodiment of the present invention, the rotation results in the actuation of multiple pins in a desired timing and sequence to control fluid flow from the cartridge to at least one channel on the chip.

[0025] In one embodiment of the present invention, the method further comprises utilizing the elasticity of the membrane to move the at least one pin in a second direction opposite the first direction.

[0026] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, shall prevail. Furthermore, the materials, methods, and examples are illustrative only and are not necessarily intended to be limiting.

[0027] Implementation of the method and / or system of embodiments of the present invention may involve performing or completing selected tasks manually, automatically, or a combination thereof. Furthermore, depending on the actual instrumentation and equipment of the method and / or system embodiments of the present invention, some selected tasks may be performed by hardware, software, firmware, or a combination thereof using an operating system.

[0028] For example, hardware for performing selected tasks according to embodiments of the present invention may be implemented as a chip or circuit. As software, selected tasks according to embodiments of the present invention may be implemented as a plurality of software instructions executed by a computer using any suitable operating system. In exemplary embodiments of the present invention, one or more tasks according to exemplary embodiments of the methods and / or systems described herein are performed by a data processor, such as a computing platform for executing a plurality of instructions. Optionally, the data processor includes volatile memory for storing instructions and / or data, and / or non-volatile storage, such as, for example, a magnetic hard disk and / or removable media, for storing instructions and / or data. Optionally, a network connection is also provided. Optionally, a display and / or user input devices, such as a keyboard and / or mouse, are also provided. [Brief explanation of the drawings]

[0029] Certain embodiments of the present invention are herein described, by way of example only, with reference to the accompanying drawings. Turning now to the detailed description, with specific reference to the drawings, it is emphasized that the specific matter shown is by way of example, is not necessarily to scale, and is for purposes of illustrating embodiments of the present invention. In this regard, the description taken together with the drawings will make apparent to those skilled in the art how embodiments of the present invention may be practiced.

[0030] [Figure 1] 1 is a perspective view of a qPCR system according to an exemplary embodiment of the present invention. [Figure 2] FIG. 1 is a perspective view of a qPCR system with the cover removed, according to an exemplary embodiment of the present invention. [Figure 3] 1 is a block diagram of a control device for a fluid volume according to an exemplary embodiment of the present invention; [Figure 4] 1 is a perspective view of a control device for a fluid volume according to an exemplary embodiment of the present invention; [Figure 5] 1 is a longitudinal cross-sectional view of a fluid volume control device according to an exemplary embodiment of the present invention; [Figure 6] 1 is a cross-sectional view of a minor axis of a control device for a fluid volume according to an exemplary embodiment of the present invention; [Figure 7] 10 is a flowchart of a method of using a fluid volume control device according to an exemplary embodiment of the present invention. [Figure 8] FIG. 1 is a top perspective view of a cartridge according to an exemplary embodiment of the present invention. [Figure 9] FIG. 2 is a bottom perspective view of a chip according to an exemplary embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0031] The present invention, in some embodiments thereof, relates to fluid flow, and more particularly, but not exclusively, to devices and methods for rapidly and efficiently moving small volumes of fluid.

[0032] Before describing at least one embodiment of the present invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details of construction and the arrangement of components and / or methods set forth in the following description and / or illustrated in the drawings. The invention is capable of other embodiments or of being practiced or being carried out in various ways.

[0033] In general, the devices and methods described herein accelerate the process of sample extraction and purification, as well as the subsequent thermal processes of the reverse transcription, extension, and denaturation steps of polymerase chain reaction ("PCR") that may occur on the product of sample purification. The presently described devices and methods rapidly and efficiently move small volumes of liquid, optionally multiple different liquids disposed in multiple corresponding wells, through one or more and / or a series of flow channels within a fluid-containing cartridge within a larger system, such as those used for real-time ("RT") qPCR analysis for COVID-19 testing. The solutions described herein achieve fluid / liquid movement in a desired sequence using a minimal number of simple components, very quickly circulating liquid volumes between at least two different regions within a chip in an RT-qPCR system.

[0034] Referring now to the drawings, Figure 1 is a perspective view of an RT-qPCR system 100 according to an exemplary embodiment of the present invention. In an embodiment of the present invention, a disposable cartridge 406 (shown in more detail with respect to Figures 4-8) is inserted into a slot 102 of the RT-qPCR system 100 for analysis, as described in more detail herein and in the related applications filed evenly and referenced in the Related Applications section. The RT-qPCR system 100 is intended to be fast, conveniently compact, easy to use, accurate, affordable, and scalable. An exemplary RT-qPCR system 100 may be available from Formulatrix, Inc. of Bedford, Massachusetts.

[0035] 2 is a perspective view of an RT-qPCR system 100 with the cover removed, according to an exemplary embodiment of the invention. Shown is a fluid volume control device 200, which forms a component part of the RT-qPCR system 100 and is shown and described in more detail with respect to FIGS. 3-6.

[0036] 3 is a block diagram of fluid volume control device 200, according to an exemplary embodiment of the present invention. In this embodiment, device 200 includes at least one camshaft 300 including at least one cam 302 located on the circumference of camshaft 300, a motor 304 for driving camshaft 300, and one or more pins 306 driven by at least one cam 302 of camshaft 300. It should be understood that device 200, which is itself a multi-part system, is a component part of a larger qPCR system 100. Within this larger RT-qPCR system 100, fluid volume control device 200 operatively interacts with cartridge 406, as described below, to, among other things, achieve controlled fluid flow within the cartridge and the overall RT-qPCR system 100.

[0037] FIG. 4 is a perspective view of a fluid volume control device 400, an example of a fluid volume control device 200, in accordance with an exemplary embodiment of the present invention. For brevity, the device 400 of FIG. 4 will be described in conjunction with FIG. 7, which is a flowchart 700 of a method of using the fluid volume control device 200 / 400. It should be appreciated that a feature of the present invention is the ability to control a variety of different liquids contained in different fluid reservoirs / wells within a multi-channel system using only a single axis of rotation of the device 200 / 400, as described in more detail below. This allows a complete sample-to-answer sequence of sample processing to occur with minimal mechanical complexity for actuating or driving the fluids within the system, using membranes positioned above all of the actuating fluids in the system.

[0038] In an embodiment of the invention, a camshaft 402 having one or more cams 408 is provided, and the camshaft 402 is rotated (702) by a motor 404 about a primary axis of rotation of the device 400. In one embodiment of the invention, the motor 404 is a stepper motor. The camshaft 402 and at least one of the cams 408 are operatively arranged such that, as the camshaft 402 is rotated by the motor 404, the at least one cam 408 contacts (704) at least one pin 412 and drives (706) the at least one pin 412 into a well 414 of the lower cartridge 406. In one embodiment of the invention, the at least one pin 412 is operatively positioned in a desired position relative to the respective cam 408 by a slot in a pin frame 410 that retains the pin 412 therein and therethrough. During actuation (706), the flexible and / or elastic membrane 500 (shown and described in more detail with respect to FIG. 5) is forced (708) into the well 414 forming a fluid seal between the membrane 500 and the wall of the well 414, thereby forcing (710) fluid in the well 414 by pressure (e.g., air pressure) into and through a chip 900 (see, e.g., channel 902 of chip 900 in FIG. 9) positioned below the cartridge 406.

[0039] It should be appreciated that as the camshaft 402 rotates and at least one cam 408 rotates to contact / drive at least one corresponding pin 412, different wells 414 of the cartridge 406 are "activated" by the depression (710) of at least one pin 412 / membrane 500. In some embodiments of the present invention, there are multiple pins 412 in the system 400 corresponding to multiple wells 414 in the cartridge 406, and rotation of the camshaft 402 about its axis of rotation, in conjunction with deliberate shapes of the cams 408 on the camshaft, results in activation of the pins at a desired timing and / or sequence, allowing multiple fluids located in the wells 414 to be precisely introduced into the channels 902 of the chip 900 for rapid and automated qPCR analysis.

[0040] In some embodiments of the invention, at least one pin 412 is spring loaded or biased to return to a resting, pre-actuated configuration when the pin 412 is not actuated (706). Optionally, the resilience / elasticity of the membrane 500 provides this spring-like behavior to the at least one pin 412. In some embodiments of the invention, the rotation (702) to pushing (710) is repeated (712), e.g., using additional cams 408 by maintaining rotation of the camshaft 400, to push additional fluid in additional wells, as desired, until all fluid wells 414 have been actuated.

[0041] In one embodiment of the present invention, using the devices and methods described herein, at least one fluid volume is driven across multiple different types of regions (within chip 900), such as at least one region that is heated to a desired temperature to achieve PCR amplification and / or at least one region that is subjected to a magnetic force (e.g., to capture the sample to be tested). Additionally, alternatively and / or optionally, at least a portion of a wash fluid is driven through the magnetically captured sample and / or an elution buffer is driven through at least one magnetized region to elute the magnetically captured sample from chip 900 or from components of the chip.

[0042] Figure 5 is a longitudinal cross-sectional view of fluid volume control device 400, according to an exemplary embodiment of the invention. Shown in Figure 5 is movement 502 of pin 412 as cam 408 drives pin 412 in a first direction, e.g., downward (as viewed from the perspective), into membrane 500, thus depressing membrane 500 into well 414, and then pin 412 moves in a second direction, e.g., upward, optionally due to the resilience of membrane 500 and / or due to being spring-loaded. As described elsewhere herein, insertion of membrane 500 into well 414 increases pressure within well 414, while removal of membrane 500 from well 414 decreases pressure within the well.

[0043] 6 is a cross-sectional view along the minor axis of fluid volume control device 400, in accordance with an exemplary embodiment of the present invention. Rotation 600 of camshaft 402 is shown, and cam 408 is biased for movement in the direction of rotation 600 in an embodiment of the present invention. As cam 408i rotates about pin 412, it drives pin 412 downward, through a membrane (not shown), and into well 414 of cartridge 406.

[0044] Figure 8 is a top perspective view of cartridge 406 according to an exemplary embodiment of the invention. Figure 9 is a bottom perspective view of chip 900 mounted within cartridge 406, showing an exemplary configuration of channel 902 according to an exemplary embodiment of the invention.

[0045] The terms "comprises," "comprising," "includes," "including," "having," and combinations thereof mean "including but not limited to."

[0046] The term "consisting of" means "including and limited to." The term "consisting essentially of" means that a composition, method, or structure may include additional components, steps, and / or moieties, but only if the additional components, steps, and / or moieties do not materially alter the basic and novel characteristics of the claimed composition, method, or structure.

[0047] The term "plurality" means "two or more."

[0048] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include multiple compounds, including mixtures thereof.

[0049] Throughout this application, various embodiments of the present invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the present invention. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, and 3 to 6, as well as individual numerical values ​​within that range, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0050] Whenever a range of values ​​is given herein, it is meant to include the cited numbers (fractions or integrals) within the range given. As used herein, the terms "range between" a first indicated value and a second indicated value, and "range from" a first indicated value to a second indicated value, are used interchangeably and are meant to include the first and second indicated values, and all fractional and integral numbers therebetween.

[0051] It will be understood that certain features of the invention that are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as appropriate in other described embodiments of the invention. Certain features described in the context of various embodiments are not considered essential features of those embodiments, unless the embodiment is inoperable without those elements.

[0052] While the present invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, the present invention is intended to embrace all such alternatives, modifications, and variations that fall within the spirit and broad scope of the appended claims.

[0053] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated herein by reference. Furthermore, citation or identification of a reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent section headings are used, they should not be construed as necessarily limiting.

Claims

1. A motor; a camshaft connected to the motor by a rotation axis of the camshaft; At least one cam disposed on the outer periphery of the camshaft; Pin frame and at least one pin disposed on said pin frame and operatively associated with at least one of said cams; a cartridge including a flexible, elastic membrane; the membrane is disposed between the cartridge and at least one of the pins; rotation of the camshaft by the motor brings at least one of the cams into contact with at least one of the pins and drives the at least one pin in a first direction; At least one of the pins is spring-biased by the flexible, resilient membrane; A device for controlling a fluid volume, wherein at least one of the pins is movable in a second direction opposite to the first direction by the flexible elastic membrane.

2. 2. The apparatus of claim 1, wherein said camshaft comprises a plurality of said cams and a plurality of said pins, each of said plurality of cams corresponding to one of said plurality of pins.

3. 3. The apparatus of claim 2, wherein the plurality of cams are arranged circumferentially about the camshaft such that rotation of the camshaft about the axis of rotation utilizes each of the plurality of cams to drive a corresponding one of the pins, thereby driving the plurality of pins in a desired timing and sequence.

4. 10. The device of claim 1, wherein the cartridge includes at least one well formed therein corresponding to at least one of the pins.

5. The apparatus of claim 1 , wherein the pin frame comprises at least one slot through which at least the pin passes.

6. 6. The apparatus of claim 5, wherein the slot in the pin frame positions at least one of the pins above a well in a cartridge, and the at least one pin is positioned between the cartridge and at least one of the cams.

7. 1. A system for performing real-time qPCR analysis, comprising: The fluid volume control device according to claim 1; a cartridge comprising a membrane and at least one well; A system comprising: a chip, wherein at least one of the wells of the cartridge is disposed between the membrane and the chip.

8. The system of claim 7 , wherein said camshaft comprises a plurality of said cams and a plurality of said pins, each of said plurality of cams corresponding to one of said plurality of pins.

9. 9. The system of claim 8, wherein the cams are arranged circumferentially about the camshaft such that rotation of the camshaft about the axis of rotation utilizes each of the cams to drive a corresponding pin, thereby driving the pins in a desired timing and sequence.

10. The system of claim 8 , wherein at least one of the cams drives at least one of the pins into at least one of the wells.

11. The system of claim 7, wherein the membrane is positioned between at least one of the pins and at least one of the wells, and when at least one of the pins is driven into the well, the membrane forms a fluid seal with the well.

12. 8. The system of claim 7, wherein at least one of the pins is provided with movement in a second direction opposite the first direction by the flexible, resilient membrane.

13. 1. A method for controlling fluid volume in a real-time qPCR system, comprising: Rotating the camshaft around a rotation axis by a motor; contacting at least one pin with a cam on the camshaft; Driving at least one of the pins in a first direction with the cam; depressing a membrane with at least one of said pins within a well of a cartridge; and using at least one of said pin and said membrane to expel fluid in said well.

14. 14. The method of claim 13, further comprising maintaining rotation to drive at least one additional pin with at least one of the cams and to push additional fluid within additional wells with the additional pin and membrane.

15. 15. The method of claim 14, wherein the rotation results in actuation of a plurality of the pins in a desired timing and sequence to control the flow of the fluid from the cartridge to at least one channel on a chip.

16. The method of claim 13 , further comprising using the elasticity of the membrane to move at least one of the pins in a second direction opposite the first direction.

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