How to pierce a seal for sample testing

Abstract

Disclosed herein include methods, compositions, and kits suitable for use in sample testing. The sample testing system can include a cartridge body for receiving a sample, at least one reaction chamber coupled to the cartridge body, and at least one seal between the cartridge body and the at least one reaction chamber. The sample testing system can include a sample dispensing mechanism operable to rupture the at least one seal to dispense a predetermined subvolume of the sample fluid from the cartridge body into the at least one reaction chamber. The sample dispensing mechanism can include a dispensing rod including at least one piercing tip that ruptures the at least one seal by forming at least one opening in the at least one seal. The piercing tip can include a geometry configured to generate a large opening in the at least one seal.

Description

[Technical field]

[0001] Related Applications This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Patent Application No. 63 / 241,033, filed September 6, 2021, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] The present disclosure relates generally to the field of sample preparation and testing, and more specifically to methods, compositions, systems, and devices for preparing and testing biological samples to aid in, for example, environmental, agricultural, scientific, veterinary, or medical diagnostics based on detecting the presence or absence of particular analytes in a sample and / or determining their quantities in a sample. [Background technology]

[0003] Nucleic acid amplification is important in many fields including medical, biomedical, environmental, veterinary, and food safety testing. Nucleic acids can be amplified by polymerase chain reaction (PCR) or isothermal amplification. After DNA amplification, there will be multiple copies of the target genetic configuration in the test solution. In diagnostic test assays, specific markers can be designed that will link to the target configuration and provide an optical signal or optical change that can be detected outside the test tube when conjugated. This optical signal can be a change in color and / or opacity of the sample measured by a change in the optical absorption of the sample at a specific optical wavelength. The output signal can also be from a direct light output from the sample that triggers the emission of a bioluminescent light output when the marker is activated by a target conjugation event. The optical detection output can also be from a change in the fluorescence of the solution, which can be from a fluorescent marker beacon. In this case, each marker molecule can be configured with a fluorescent quencher in close proximity to a fluorescent atom or configuration of atoms. The marker molecule can be configured such that when it selectively binds to a target DNA location within the test solution, the quencher and fluorophore are separated, and a strong fluorescent signal can then be detected by the action of the fluorophore. In this configuration, the overall fluorescence intensity of the target solution is indicative of the relative amount of target genetic material within the test solution. This signal can then be used to form the basis of a diagnostic test to determine the presence, absence, and relative amount of target material within a sample under test. Summary of the Invention [Problem to be solved by the invention]

[0004] Current sample testing systems and devices, particularly nucleic acid amplification and detection instruments, require sample preparation steps that are typically large, complex, and expensive and must be performed independently of the instrument. These preparation steps typically require a trained technical operator who and the test preparation environment can be exposed to hazardous samples such as bodily fluids and infectious agents, and the process is subject to risks from incorrect manual operations including spills and incorrect reagent additions. The resulting test sample must then be accurately subsampled and transferred in a manual transfer step, typically by skilled pipetting operations. This approach requires a trained technical operator and several separate tubes and transfer devices, all of which will become contaminated by the sample and must be properly handled and individually discarded. In these approaches, the test sample is not sealed from the environment during the process of sample preparation and transfer into the test tube in the test instrument. This exposure to the sample can pose an infectious agent risk to the user and others, and can also contaminate the test instrument and testing area, resulting in incorrect diagnostic results in subsequent tests.

[0005] An alternative approach involves the cartridge body (e.g., a sample preparation reservoir) securely holding the sample preparation solution therein until such time that a partial volume (e.g., a predetermined partial volume of sample fluid from the cartridge body) is dispensed through a perforation in an otherwise sealed wall between the cartridge body and a coupled reaction chamber (e.g., a test tube). However, to achieve the desired chemical performance, the amount of sample fluid dispensed from the cartridge body into the reaction chamber must be accurate within small tolerances. A need exists for methods, configurations, systems, and devices that improve the flow and accuracy of sample fluid dispensed from the cartridge body into this reaction chamber. [Means for solving the problem]

[0006] Disclosed herein includes a sample inspection system. In some embodiments, the sample inspection system includes a cartridge body for receiving a biological or environmental sample for preparation of a sample fluid therefrom in a sample preparation fluid contained in the cartridge body. In some embodiments, the sample inspection system includes at least one reaction chamber coupled to the cartridge body. In some embodiments, the sample inspection system includes at least one seal between the cartridge body and the at least one reaction chamber to prevent fluid transfer between the cartridge body and the at least one reaction chamber. In some embodiments, the sample inspection system includes a sample dispensing mechanism for insertion into the cartridge body (e.g., after receiving a biological or environmental sample therein). The sample inspection system can include at least one seal to prevent fluid transfer between the cartridge body and the at least one reaction chamber. In some embodiments, the at least one seal is located between the cartridge body and the at least one reaction chamber. In some embodiments, the at least one seal can be pierced by a piercing tip, thereby allowing fluid transfer between the cartridge body and the at least one reaction chamber.

[0007] In some embodiments, the sample dispensing mechanism is operable to rupture at least one seal to allow sample fluid to flow from the cartridge body into the at least one reaction chamber, and to dispense a predetermined subvolume of sample fluid from the cartridge body into the at least one reaction chamber for testing therein while preventing further fluid transfer between the cartridge body and the at least one reaction chamber. In some embodiments, the sample dispensing mechanism includes a dispensing rod including at least one piercing tip that ruptures the at least one seal by forming at least one opening therein, the at least one piercing tip including a geometry configured to generate a large opening in the at least one seal. In some embodiments, the cartridge body provides an initially unobstructed open volume such that a swab carrying a biological or environmental sample can be used to agitate the sample preparation fluid in the cartridge body and to flush the biological or environmental sample from the swab into the sample preparation fluid.

[0008] The sample inspection system may include a closure for sealing the cartridge body and the sample dispensing mechanism therein after receipt of a biological or environmental sample. In some embodiments, at least one of the closure and the cartridge body is configured to prevent or at least inhibit removal of the closure from the cartridge body such that fluids remain sealed within the sample inspection system. In some embodiments, the sample dispensing mechanism is attached to the closure such that the act of adding the closure to the cartridge body also accomplishes insertion of the sample dispensing mechanism into the cartridge body. The seal may prevent fluid transfer prior to seal puncture. In some embodiments, the shape of the cartridge body prevents rotation when screwing on the cap and orients the multiple reaction chambers in a specific position. In some embodiments, a single action by a user causes the sample dispensing mechanism to rupture at least one seal and dispense sample fluid from the cartridge body into at least one reaction chamber. In some embodiments, the single action by a user is a sustained screwing action applied to the closure against the cartridge body, the screwing action causing actuation of the sample dispensing mechanism and sealing the cartridge body. In some embodiments, the closure includes a screw thread. The sample testing system may include a second closure that seals the sample preparation fluid within the cartridge body prior to use and is removed to allow a biological or environmental sample to be added to the sample preparation fluid contained in the cartridge body. In some embodiments, a single action by a user is a downward force applied to the closure against the cartridge body. In some embodiments, the downward force forms a snap fit between the closure and the cartridge body. In some embodiments, the downward force causes actuation of a sample dispensing mechanism and seals the cartridge body. In some embodiments, the downward force includes a downward force of a lever means. In some embodiments, the closure includes a snap-on dispensing cap including one or more snap-on members configured to form a snap fit with a distal end of the cartridge body upon a single action by a user.

[0009] In some embodiments, the sample dispensing mechanism includes a dispensing chamber that forms a second seal against the at least one seal to capture a predetermined partial volume of sample fluid in the dispensing chamber, and a plunger mechanism that forms a sliding seal with an inner surface of the dispensing chamber, the sliding seal configured to slide along the inner surface of the dispensing chamber to dispense the predetermined partial volume of sample fluid therefrom through the at least one opening and into the at least one reaction chamber. In some embodiments, the dispensing chamber includes an outer surface having spaced apart chamber positioning features extending therefrom and configured to center the dispensing chamber in the cartridge body to allow sample fluid to flow between the chamber positioning features when the sample dispensing mechanism is inserted into the cartridge body.

[0010] In some embodiments, the sample dispensing mechanism is configured such that a single action performed by a user triggers two stages of operation of the sample dispensing mechanism including a first stage of operation in which a predetermined partial volume of sample fluid is captured in the dispensing chamber and a second stage of operation in which the sample fluid is dispensed from the dispensing chamber. In some embodiments, the sample dispensing mechanism includes a force prioritization component that is reconfigured or destroyed to allow the second stage of operation. In some embodiments, the force prioritization component includes a breakable component configured to break to allow operation of the sample dispensing mechanism to proceed from the first stage of operation to the second stage of operation. In some embodiments, the force prioritization component includes a collapsible or crushable spacer that presses against the dispensing chamber to seal it in the first stage of operation and that is crushed or squashed in the second stage of operation to maintain the seal, perform the piercing action, and actuate the plunger so that the sample fluid is dispensed from the dispensing chamber.

[0011] In some embodiments, at least one penetrating tip comprises a ball point tip. In some embodiments, at least one penetrating tip comprises an arrowhead tip. In some embodiments, at least one penetrating tip comprises a frusto-conical tip. In some embodiments, at least one penetrating tip does not comprise a sharp tip. In some embodiments, a distal portion of at least one penetrating tip comprises a flat surface. In some embodiments, the flat surface forms an angle of less than about 20°, about 15°, about 10°, about 5°, or about 1° with respect to a surface of the at least one seal.

[0012] In some embodiments, the at least one piercing tip is fluted. In some embodiments, the at least one piercing tip includes one or more flow channels. In some embodiments, the one or more flow channels are positioned (i) at the proximal end of the at least one piercing tip, (ii) at the distal end of the at least one piercing tip, or (iii) over the length of the at least one piercing tip. In some embodiments, at least a portion of the predetermined subvolume of the sample fluid flows through the at least one opening through the one or more flow channels. In some embodiments, the fluid flow is at a high flow rate compared to a sample inspection system in which the at least one piercing tip does not include one or more flow channels. In some embodiments, the one or more flow channels include a longitudinal groove extending along the at least one piercing tip. In some embodiments, the step of the at least one piercing tip rupturing the at least one seal includes the step of the at least one piercing tip penetrating the at least one seal and moving into at least a portion of the at least one reaction chamber. In some embodiments, the at least one opening increases in size as the at least one piercing tip moves into at least a portion of the at least one reaction chamber, hi some embodiments, the at least one opening remains substantially the same size as the at least one piercing tip moves into at least a portion of the at least one reaction chamber.

[0013] In some embodiments, the at least one reaction chamber includes a trapped gas, and the cartridge body includes a gaseous head space above the sample fluid. In some embodiments, the dispensing rod is configured to equalize pressure between the gaseous head space and the at least one reaction chamber after the at least one seal is ruptured. In some embodiments, the at least one penetrating tip includes at least one vent opening leading to a vent lumen extending through the dispensing rod, and the dispensing rod includes a vent port positioned in the gas head space and in fluid communication with the vent lumen of the dispensing rod. In some embodiments, the sample dispensing mechanism includes at least one hydrophobic filter. In some embodiments, any fluid passing between the at least one vent port and the at least one vent opening must pass through the hydrophobic filter. In some embodiments, the vent opening is positioned (i) at a proximal end of the at least one penetrating tip, (ii) at a distal end of the at least one penetrating tip, or (iii) over the length of the at least one penetrating tip. In some embodiments, trapped gas displaced by the at least one penetration tip and / or the predetermined subvolume of the sample fluid can escape into the gas headspace through the at least one vent opening.

[0014] In some embodiments, the at least one piercing tip that ruptures the at least one seal functions to generate one or more flaps, the one or more flaps comprising a portion of the at least one seal ruptured by the at least one piercing tip. In some embodiments, the flaps do not adhere to the at least one piercing tip and / or do not disrupt fluid flow through the opening. In some embodiments, the piercing tip comprises a geometry configured to reduce wicking of sample fluid into the at least one piercing tip and / or the one or more flaps.

[0015] The large opening can include, for example, piercing at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 99% of the area of ​​the at least one seal. In some embodiments, at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 99% of the area of ​​the at least one seal is in contact with the at least one piercing tip. In some embodiments, at least about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 99% of the predetermined subvolume of the sample fluid flows into the at least one reaction chamber. In some embodiments, after at least one seal is ruptured, less than about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, or about 1% of the sample fluid of a predetermined partial volume of the sample fluid remains in or on the dispensing chamber, the at least one seal, and / or the at least one piercing tip. In some embodiments, the predetermined subvolume of the sample fluid comprises at least about 10 μL, about 15 μL, about 20 μL, about 25 μL, about 30 μL, about 35 μL, about 40 μL, about 45 μL, about 50 μL, about 60 μL, about 70 μL, about 80 μL, about 90 μL, about 100 μL, about 110 μL, about 120 μL, about 128 μL, about 130 μL, about 140 μL, about 150 μL, about 160 μL, about 170 μL, about 180 μL, about 190 μL, or about 200 μL of the sample fluid.

[0016] In some embodiments, the sample dispensing mechanism includes an overmolded layer disposed on a surface of at least a portion of the dispensing rod and / or dispensing chamber. In some embodiments, the overmolded layer forms a seal. In some embodiments, it is a cylindrical seal. In some embodiments, the overmolded layer includes a thermoplastic elastomer (TPE) having a different durometer hardness than at least a portion of the dispensing rod and / or dispensing chamber. In some embodiments, the overmolded layer exhibits a Shore D durometer hardness or a Shore A durometer hardness of about 20-30.

[0017] In some embodiments, the dispensing chamber is initially configured such that when the sample dispensing mechanism is inserted into the cartridge body, the sample fluid is forced to flow around the outside of the dispensing chamber before it can flow into the dispensing chamber, and the fluid flowing around the outside of the dispensing chamber is forced through a filter or porous filling material that retains and / or captures particles and contaminants, and / or incorporates biological or chemical components that bind to or capture sample fluid components that may otherwise inhibit or interfere with sample testing.

[0018] In some embodiments, the cartridge body contains one or more magnetic particles along with a sample preparation fluid, the surfaces of which are coated or functionalized to bind to and capture at least one predetermined target chemical species of a biological or environmental sample when mixed into the sample fluid, the sample dispensing mechanism is configured to force the sample fluid through the dispensing chamber when the sample dispensing mechanism is inserted into the cartridge body, one or more magnets are positioned in close proximity to an inner surface of the dispensing chamber such that magnetic particles contained within the sample fluid and that have captured the target chemical species are attracted to and held against the inner surface of the dispensing chamber, and a plunger mechanism that forms a sliding seal with the inner surface of the dispensing chamber thereby collects the magnetic particles held against the inner surface and dispenses them into at least one reaction chamber, generating an increase in the concentration of at least one predetermined target chemical species in a predetermined subvolume of the sample fluid dispensed therein.

[0019] In some embodiments, the at least one reaction chamber is a double reaction chamber and the at least one piercing tip is a double piercing tip. In some embodiments, the reaction chamber comprises a polymerase chain reaction (PCR) tube. In some embodiments, the two reaction chambers comprise one or more mixing beads. In some embodiments, the reaction chambers comprise different reagents selected to perform respective different tests and / or detect respective different target entities. In some embodiments, the cartridge body comprises sample preparation reagents and at least one of the reaction chambers comprises one or more reagents suitable for reverse transcription and / or amplification reactions. In some embodiments, the cartridge body comprises one or more alignment features configured to align with and engage one or more mating slots of a testing device. In some embodiments, the one or more alignment features prevent rotation of the cartridge body when the cartridge body is positioned in the testing device. In some embodiments, the one or more alignment features allow a user to remove the second closure and / or perform a single action in a one-handed operation.

[0020] Disclosed herein includes a method of sample testing. In some embodiments, the method of sample testing includes adding a biological or environmental sample into a sample preparation fluid contained in a cartridge body of a sample testing system disclosed herein for preparation of the sample in the system, and after the adding step, inserting a sample dispensing mechanism into the cartridge body and attaching a closure to the cartridge body, and rupturing at least one seal between the cartridge body and the at least one reaction chamber to allow sample fluid to flow from the cartridge body into the at least one reaction chamber, while preventing further fluid transfer between the cartridge body and the at least one reaction chamber, and actuating the sample dispensing mechanism to dispense a predetermined partial volume of the sample fluid from the cartridge body into the at least one reaction chamber for testing in the at least one reaction chamber. The method may include, before the adding step, placing the sample testing system into a receiving port of a testing device configured to perform a test on the biological or environmental sample therein.

[0021] In some embodiments, a testing device is provided. The testing device may include a receiving port configured to receive a sample testing system provided herein. In some embodiments, the testing device is configured to perform a test thereon on a biological or environmental sample. In some embodiments, the testing device further includes a lever means configured to apply a downward force to a sample testing system positioned in the receiving port. In some embodiments, the single action is to apply a downward force to the closure against the cartridge body through the lever means. In some embodiments, the downward force forms a snap-fit ​​between the closure and the cartridge body. In some embodiments, the lever means includes a hinged lid. In some embodiments, the hinged lid includes a ridge configured to contact a surface of the closure. In some embodiments, the testing device further includes a sleeve for storing the lever means. In some embodiments, the hinged lid is substantially parallel to the cartridge body when stored in the sleeve. In some embodiments, at least a portion of the hinged lid is configured to slide up and out of the sleeve when lifted by a user to expose a hinge of the hinged lid. In some embodiments, when the hinge is exposed, the hinged lid has the ability to pivot to a horizontal position substantially perpendicular to the cartridge body. In some embodiments, the testing device includes one or more mating slots configured to align with and engage with one or more alignment features of the cartridge body. In some embodiments, the one or more mating slots are located at the receiving port. In some embodiments, the one or more alignment features prevent rotation of the cartridge body when the cartridge body is positioned in the testing device. In some embodiments, the one or more alignment features allow a user to remove the second closure and / or perform a single action in a one-handed operation. [Brief description of the drawings]

[0022] [Figure 1]FIG. 1 is a non-limiting, illustrative schematic diagram of a dual reaction chamber cartridge with an insert sample dispensing mechanism (including a dual piercing tip) in an initial position. [Diagram 2] FIG. 1 is a non-limiting, illustrative schematic diagram of a dual reaction chamber cartridge with an insert sample dispensing mechanism (including a dual piercing tip) in an initial position. [Diagram 3] A non-limiting illustrative schematic diagram of the cartridge shown in Figures 1 and 2 with the sample dispensing mechanism fully inserted, such that the seal is fully penetrated by the piercing tip and a predetermined partial volume of sample fluid from the cartridge body is dispensed into the dual reaction chamber. [Figure 4] A non-limiting illustrative schematic diagram of the cartridge shown in Figures 1 and 2 with the sample dispensing mechanism fully inserted, such that the seal is fully penetrated by the piercing tip and a predetermined partial volume of sample fluid from the cartridge body is dispensed into the dual reaction chamber. [Diagram 5] FIG. 13 is a non-limiting exemplary schematic diagram of a penetrating tip ball point flow channel embodiment with an overmolded layer of a sample dispensing mechanism shown in brackets. [Figure 6] FIG. 13 is another non-limiting exemplary schematic diagram of a ball point flow channel embodiment of a penetrating tip. [Figure 7] 1A-1C are non-limiting, exemplary schematic diagrams of ball point embodiments of penetrating tips provided herein with vent openings positioned at the distal end of the penetrating tip. [Figure 8] FIG. 13 is a non-limiting, exemplary schematic diagram of a sample dispensing mechanism with vent openings positioned over the length of the penetrating tip. [Figure 9] FIG. 9 is a non-limiting, exemplary schematic diagram of a cross-sectional side view of the sample dispensing mechanism shown in FIG. 8 showing gas vent openings leading to the interior chamber. [Figure 10] 1A-1C are non-limiting, exemplary schematic diagrams of a penetrating tip with vent openings positioned over the length of the penetrating tip. [Figure 11] FIG. 13 is a non-limiting exemplary schematic diagram of a dispensing cap assembly in which a piercing tip includes a flow channel and a transition to a larger opening. [Figure 12] FIG. 13 is a non-limiting, exemplary schematic diagram of a sample dispensing mechanism with a vent opening positioned at the distal end of the penetrating tip. [Figure 13] FIG. 13 is a non-limiting, exemplary schematic diagram of a cross-sectional side view of the sample dispensing mechanism shown in FIG. 12 showing an internal vent path having a hydrophobic filter. [Figure 14] 1A-1C are non-limiting exemplary schematic diagrams of cross-sectional views of penetration rods provided herein. [Figure 15] FIG. 2 is a non-limiting, exemplary schematic diagram of a dispensing rod including a piercing tip that does not have a sharp tip. [Figure 16] FIG. 13 is a non-limiting, exemplary schematic diagram of a dispensing rod including a fluted sagittal penetrating tip. [Figure 17] FIG. 1 is a non-limiting, illustrative schematic diagram of a cartridge having two diagnostic test reservoirs (eg, reaction chambers). [Figure 18] FIG. 18 is a non-limiting, exemplary schematic diagram of the dual reaction chamber cartridge of FIG. 17. [Figure 19] A non-limiting illustrative schematic diagram of the cartridge of Figures 17 and 18 with the shipping cap removed and a swab inserted into the open volume of the cartridge body of the dual reaction chamber cartridge to allow sample material to accumulate in this volume. [Figure 20] FIG. 2 is a non-limiting illustrative schematic diagram of a cap assembly including a cap and an embodiment of a dual reaction chamber dispensing mechanism immediately prior to insertion into a cartridge body of a dual reaction chamber cartridge. [Figure 21] FIG. 1 is a non-limiting, exemplary schematic diagram of a dual reaction-chamber dispensing mechanism in which the dispensing chamber (e.g., dispensing insert) is separated from the dispensing rod. [Figure 22] FIG. 1 is a non-limiting, exemplary schematic diagram of a cross-sectional side view of a dual reaction chamber cartridge with a dispense mechanism partially inserted. [Diagram 23] A non-limiting, illustrative schematic diagram of a side cross-sectional view of a dual reaction chamber cartridge with a dispense mechanism further inserted, with the dispense chamber of the dispense mechanism seated against the base of the cartridge, but prior to piercing and dispensing. [Figure 24] FIG. 2 is a non-limiting, exemplary schematic diagram of a cross-sectional side view of a dual reaction chamber cartridge with the dispense mechanism fully inserted, thus completing the piercing and dispense action. [Diagram 25] FIG. 25 is a non-limiting exemplary schematic diagram of an exterior view of a dual reaction chamber cartridge after fully engaging the cap assembly as in FIG. 24. [Figure 26] 1A-1C are non-limiting, illustrative schematic diagrams of fluid flow, including mixing, through and around the dispensing insert as the dispensing mechanism is pressed into the cartridge. [Figure 27] FIG. 2 is a non-limiting, illustrative schematic diagram of an isolated dispense chamber. [Figure 28] A non-limiting exemplary schematic diagram of a side cross-sectional view showing an alternative embodiment of a dispensing chamber in which when the base of the insert bore is sealed and the insert is pressed into the cartridge, the sample and sample reagent fluids can only flow through the outer bypass area of ​​the insert and then flow back to the top of the insert bore, the bypass area may include a filter, porous material, or filling material to remove or capture any particles in the sample fluid and prevent these particles from flowing into the test reservoir. [Figure 29] FIG. 13 is a non-limiting, illustrative schematic diagram of a cross-sectional side view showing yet another alternative embodiment of a dispensing chamber in which sample fluid can only flow through the internal bore of the insert when the insert assembly is pressed into the cartridge, the bypass area is blocked so that the sample fluid cannot bypass the insert barrel, and the insert includes a magnet around the bore for collecting and concentrating DNA and RNA captured on the surface of the magnetic beads. [Figure 30A] A non-limiting exemplary schematic diagram of a top view of a snap-on dispensing cap assembly and a dual reaction chamber cartridge in an initial position before a snap fit is formed between the cap assembly and the dual reaction chamber cartridge. [Figure 30B]A non-limiting illustrative schematic diagram of an oblique view of a snap-on dispensing cap assembly and cartridge in an initial position before a snap fit is formed between the cap assembly and dual reaction chamber cartridge, with the arrow indicating the downward force applied by the user to engage the snap. [Figure 30C] A non-limiting exemplary schematic diagram of a side view of a snap-on dispensing cap assembly and a dual reaction chamber cartridge in an initial position before a snap fit is formed between the cap assembly and the dual reaction chamber cartridge. [Figure 30D] A non-limiting exemplary schematic diagram of a side view of a snap-on dispensing cap assembly and a dual reaction chamber cartridge in an initial position before a snap fit is formed between the cap assembly and the dual reaction chamber cartridge. [Figure 31A] FIG. 30C is a non-limiting, exemplary schematic diagram of a top view of the snap-on dispensing cap assembly shown in FIGS. 30A-30D and the dual reaction chamber cartridge after it has been fully engaged to form a snap fit. [Figure 31B] A non-limiting exemplary schematic diagram of a perspective view of the snap-on dispensing cap assembly shown in Figures 30A-30D with the arrow indicating the snap-on and the dual reaction chamber cartridge after it has been fully engaged therewith to form a snap-on. [Figure 31C] FIG. 30C is a non-limiting, exemplary schematic diagram of a side view of the snap-on dispensing cap assembly shown in FIGS. 30A-30D and the dual reaction chamber cartridge after it has been fully engaged to form a snap fit. [Figure 31D] FIG. 30C is a non-limiting, exemplary schematic diagram of a side view of the snap-on dispensing cap assembly shown in FIGS. 30A-30D and the dual reaction chamber cartridge after it has been fully engaged to form a snap fit. [Figure 32A] A non-limiting exemplary schematic diagram of a perspective view of a testing device with a snap-on dispensing cap assembly and a dual reaction chamber cartridge (in an initial position before a snap-fit ​​is formed between them) positioned in a receiving port. [Figure 32B]A non-limiting, exemplary schematic diagram of a side view of a testing device with a snap-on dispensing cap assembly and a dual reaction chamber cartridge (in an initial position before a snap fit is formed between them) positioned in a receiving port. [Figure 33A] A non-limiting exemplary schematic diagram of a perspective view of the testing device shown in Figures 32A-32B after the hinged lid has been pressed onto the top of the snap-on dispensing cap to form a snap fit, with an arrow indicating where the user can press on the hinged lid to engage the snap. [Figure 33B] 32A-32B after the hinged lid has been pressed onto the top of the snap-on dispensing cap to form a snap fit. FIG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In these drawings, similar symbols generally identify similar components unless the context dictates otherwise. The exemplary embodiments described in the detailed description, drawings, and claims are not intended to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described and illustrated in the figures herein, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are expressly contemplated herein and made a part of the present disclosure herein.

[0024] All patents, published patent applications, other literature, and sequences from GenBank and other databases referenced herein are incorporated by reference in their entirety for the relevant art.

[0025] Disclosed herein includes a sample inspection system. The sample inspection system can include a cartridge body for receiving a biological or environmental sample into a sample preparation fluid contained in the cartridge body for preparation of the sample fluid from the cartridge body. In some embodiments, the sample inspection system includes at least one reaction chamber coupled to the cartridge body. In some embodiments, the sample inspection system includes at least one seal between the cartridge body and the at least one reaction chamber to prevent fluid transfer between the cartridge body and the at least one reaction chamber. In some embodiments, the sample inspection system includes a sample dispensing mechanism for insertion into the cartridge body after receiving the biological or environmental sample into the cartridge body.

[0026] In some embodiments, the sample dispensing mechanism is operable to rupture at least one seal to allow sample fluid to flow from the cartridge body into the at least one reaction chamber, and dispense a predetermined subvolume of sample fluid from the cartridge body into the at least one reaction chamber for testing in the at least one reaction chamber while preventing further fluid movement between the cartridge body and the at least one reaction chamber. In some embodiments, the sample dispensing mechanism includes a dispensing rod including at least one piercing tip that ruptures the at least one seal by forming at least one opening therein, the at least one piercing tip including a geometry configured to generate a large opening in the at least one seal. In some embodiments, the cartridge body provides an initially unobstructed open volume to allow a swab carrying a biological or environmental sample to be used to agitate the sample preparation fluid within the cartridge body and to flush the biological or environmental sample from the swab into the sample preparation fluid.

[0027] Disclosed herein includes a method of sample testing. The method of sample testing may include, for example, adding a biological or environmental sample into a sample preparation fluid contained in a cartridge body of a sample testing system disclosed herein for preparation of the sample in the system, and after the adding step, inserting a sample dispensing mechanism into the cartridge body and attaching a closure to the cartridge body, and rupturing at least one seal between the cartridge body and the at least one reaction chamber to allow sample fluid to flow from the cartridge body into the at least one reaction chamber, while preventing further fluid transfer between the cartridge body and the at least one reaction chamber, and actuating the sample dispensing mechanism to dispense a predetermined partial volume of the sample fluid from the cartridge body into the at least one reaction chamber for testing in the at least one reaction chamber. The method may include, before the adding step, placing the sample testing system into a receiving port of a testing device configured to perform a test on the biological or environmental sample therein.

[0028] Unless otherwise defined, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. See, for example, Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons, NY, USA, 1994; Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, NY, USA, 1989. For purposes of this disclosure, the following terms are defined:

[0029] In some embodiments, methods, configurations, systems, and devices for sample testing are provided. Disclosed herein include methods, configurations, and systems for penetrating seals for patient sample assays and penetrating sample assays. In some embodiments, a dual rod penetrating and fluid dispensing device is provided. The device can be part of a consumable for a molecular point-of-care rapid testing system. The consumable device can include a two-part system. The first part of the system can include a main cartridge that includes a cap and an upper section (e.g., sample preparation reservoir) called the cartridge body filled with diluent (sample preparation fluid), and a lower section called the dual tube, which is a symmetrical reaction chamber pair each containing dried molecular reagents and mixing beads, with the upper and lower sections separated by a foil seal and an elastomeric gasket. The second part of the system can include a dispensing rod and / or a cap assembly.

[0030] In some embodiments of the uses provided herein, a patient sample collected using a swab is generally mixed into a diluent in the cartridge. The swab can be removed and then a dispensing rod can be inserted into the diluent in the cartridge. In some embodiments, the step of screwing the cap and cap assembly of the dispensing rod drives the dispensing rod downward through the cartridge body, piercing the foil seals just above each of the two reaction chambers, and driving the sample fluid through the seals and into the reaction chambers. To achieve the desired chemical performance, the amount of fluid dispensed into the reaction chambers must be accurate within a small tolerance.

[0031] In some embodiments, a dispensing rod configured to improve the flow and accuracy of dispensing fluid is provided. Provided herein are various piercing tip geometries with the central goal of generating larger openings in the foil and larger volumetric pathways through which to direct liquid flow.

[0032] Currently available configurations and methods suffer from cartridge design shortcomings where liquid is forced into a sealed reaction chamber, compressing the gas in the chamber and creating pressure against the seal, increasing the chance of leakage, and thereby reducing the accuracy of the dispensed volume. The embodiments provided herein contemplate various geometries that allow the gas in the reaction chamber to vent into the headspace of the cartridge body to equalize the pressure, thereby improving the reliability of the seal and the accuracy of the dispensed volume. In some embodiments, a composite plastic disposable is provided herein that pierces the foil and pushes the patient sample towards the assay, delivering and dispensing a predetermined subvolume (e.g., 100 uL) of the sample fluid from the cartridge body into at least one reaction chamber of the reagent dual tube region.

[0033] In some embodiments, the piercing tip geometry is such that a maximum opening is provided that allows maximum dispensing into the reaction chamber. In some embodiments, the piercing tip geometry is also designed to limit wicking of the aliquot and / or vent trapped gas in the reagent chamber. The embodiments of the dispensing rod and the piercing tip provided herein can be combined, for example, a geometry can be added that allows venting of trapped gas to increase the accuracy of the dispensed aliquot. In some embodiments, the tip geometry provided herein allows for a larger piercing orifice suitable for more precise volume dispensing as well as possible venting of trapped gas in the closed chamber.

[0034] A sharp tip may be able to pierce a seal such as an AL seal, but close back around the shaft, causing poor flow and fluid adhesion. The ball point tip of the piercing tip described herein may allow for a larger puncture, increasing fluid flow and preventing the tendency of seals such as foils to become trapped on the shaft of the dispensing rod. The fluted tip design of the piercing tip described herein may allow for a larger puncture while allowing for venting and allowing fluid to flow through the flutes. The arrowhead design of the piercing tip described herein may improve fluid flow and increase the accuracy of the dispensed aliquot.

[0035] Provided herein are foil-piercing rods with unique geometries to improve fluid dispensing and dual-piercing dispensing rods for volume consistency. In some embodiments, PCR foil dispensing rods are provided. The methods, configurations, systems, and devices provided herein, such as the dispensing rods disclosed herein, can be used in a variety of dispensing situations other than PCR tubes. The configurations, systems, and methods described herein are useful in a variety of different environments where a predetermined partial volume of a sample fluid is dispensed into a chamber (e.g., a reaction chamber).

[0036] Without being bound by any particular theory, the methods, configurations, systems, and devices disclosed herein provide improved performance compared to currently available methods and systems due to the improved fluid flow, reduced fluid adhesion, and / or venting of trapped gas in the embodiments provided herein.

[0037] Disclosed herein includes a sample inspection system. The sample inspection system may include a cartridge body for receiving a biological or environmental sample into a sample preparation fluid contained in the cartridge body for preparation of the sample fluid from the cartridge body. The sample inspection system may include at least one reaction chamber coupled to the cartridge body. The sample inspection system may include at least one seal between the cartridge body and the at least one reaction chamber to prevent fluid transfer between the cartridge body and the at least one reaction chamber. The sample inspection system may include a sample dispensing mechanism for insertion into the cartridge body after receiving the biological or environmental sample into the cartridge body.

[0038] In some embodiments, the sample dispensing mechanism can be operable to rupture at least one seal to allow sample fluid to flow from the cartridge body into the at least one reaction chamber, and dispense a predetermined sub-volume of sample fluid from the cartridge body into the at least one reaction chamber for testing in the at least one reaction chamber while preventing further fluid movement between the cartridge body and the at least one reaction chamber. The sample dispensing mechanism can include a dispensing rod including at least one piercing tip that ruptures the at least one seal by forming at least one opening in the at least one seal. The at least one piercing tip can include a geometry configured to generate a large opening in the at least one seal. The large opening can include puncturing of at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% of the area of ​​the at least one seal, or a number or range between any two of these values. In some embodiments, the cartridge body provides an initially unobstructed open volume so that a swab carrying a biological or environmental sample can be used to agitate the sample preparation fluid within the cartridge body and wash the biological or environmental sample off the swab and into the sample preparation fluid.

[0039] The sample dispensing mechanism may include a dispensing chamber that forms a second seal against the at least one seal to capture a predetermined partial volume of sample fluid in the dispensing chamber. The sample dispensing mechanism may include a plunger mechanism that forms a sliding seal with an inner surface of the dispensing chamber configured to slide along the inner surface of the dispensing chamber to dispense a predetermined partial volume of sample fluid from the dispensing chamber through the at least one opening into the at least one reaction chamber. In some embodiments, the dispensing chamber may include an outer surface having spaced apart chamber positioning features extending from an outer surface thereof and configured to center the dispensing chamber in the cartridge body, the chamber positioning features configured to allow sample fluid to flow between the chamber positioning features when the sample dispensing mechanism is inserted into the cartridge body.

[0040] The sample dispensing mechanism may be configured such that a single action performed by a user triggers two stages of operation of the sample dispensing mechanism including a first stage of operation in which a predetermined partial volume of sample fluid is captured in the dispensing chamber and a second stage of operation in which sample fluid is dispensed from the dispensing chamber. The sample dispensing mechanism may include a force prioritization component that is reconfigured or destroyed to enable the second stage of operation. The force prioritization component includes a breakable component configured to break to enable operation of the sample dispensing mechanism to proceed from the first stage of operation to the second stage of operation. The force prioritization component may include a collapsible or crushable spacer that presses against the dispensing chamber to seal the dispensing chamber in the first stage of operation and that is crushed or squashed to maintain the seal and perform a piercing action to actuate the plunger such that sample fluid is dispensed from the dispensing chamber in the second stage of operation.

[0041] At least one of the piercing tips may include a ball-point tip. At least one of the piercing tips may include an arrowhead tip. At least one of the piercing tips may include a truncated cone tip. Figures 1 and 2 depict non-limiting, exemplary schematic diagrams of a dual reaction chamber cartridge with an inserted sample dispensing mechanism 50 (including a dual piercing tip 52) in an initial position. The dual reaction chamber 54 may be joined to a cartridge body 56 provided herein. A seal 58 between the cartridge body and the reaction chamber may prevent fluid transfer between the cartridge body and the reaction chamber. Figures 3 and 4 depict non-limiting, exemplary schematic diagrams of the cartridge shown in Figures 1 and 2 with the sample dispensing mechanism fully inserted such that the seal 58 is fully pierced by the piercing tip and a predetermined partial volume of sample fluid from the cartridge body 56 is dispensed into the dual reaction chamber 54.

[0042] Some embodiments of the sample inspection system provided herein include one or more overmolded layers. One or more components of the cartridge provided herein may include a thermoplastic elastomer (TPE). Provided herein are cartridges that include one or more overmolded layers of different durometer hardness. The selection of the TPE and its durometer hardness may vary depending on the embodiment and the nature and purpose of the overmolded layer. In some embodiments, the sample dispensing mechanism includes an overmolded layer disposed on a surface of at least a portion of the dispensing rod and / or dispensing chamber. The overmolded layer may form a seal, such as a cylindrical seal. The overmolded layer may include a thermoplastic elastomer (TPE) different from the surface that the overmolded layer covers, such as at least a portion of the dispensing rod and / or dispensing chamber. The durometer hardness of the TPE may vary in various embodiments. In some embodiments, the durometer hardness of the TPE is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or a number or range between or about any two of these values. In some embodiments, the durometer hardness of the TPE is, on the Shore A scale or Shore D, min or max 1, min or max 2, min or max 3, min or max 4, min or max 5, min or max 6, min or max 7, min or max 8, min or max 9, min or max 10, min or max 11, min or max 12, min or max 13, min or max 14,min or max 15, min or max 16, min or max 17, min or max 18, min or max 19, min or max 20, min or max 21, min or max 22, min or max 23, min or max 24, min or max 25, min or max 26, min or max 27, min or max 28, min or max 29, min or max 30, min or max 31, min or max 32, min or max 33, min or max 34, min or max 35, min or max 36, min or max 37, min or max 38, min or max 39, min or max 40, min or max 41, min or max 42, min or max 43, min or max 44, min or max 45, min or max 46, min or max 47, min or max 48, min or max 49, min or max 50, min or max 51, min or max 52, min or max 53, min or max 54, min or max 55, min or max 56, min or max 57, min or max 58 for maximum, 59 for minimum or maximum, 60 for minimum or maximum, 61 for minimum or maximum, 62 for minimum or maximum, 63 for minimum or maximum, 64 for minimum or maximum, 65 for minimum or maximum, 66 for minimum or maximum, 67 for minimum or maximum, 68 for minimum or maximum, 69 for minimum or maximum, 70 for minimum or maximum, 71 for minimum or maximum, 72 for minimum or maximum, 73 for minimum or maximum, 74 for minimum or maximum, 75 for minimum or maximum, 76 for minimum or maximum, 77 for minimum or maximum, 78 for minimum or maximum, 79 for minimum or at most 80, at least or at most 81, at least or at most 82, at least or at most 83, at least or at most 84, at least or at most 85, at least or at most 86, at least or at most 87, at least or at most 88, at least or at most 89, at least or at most 90, at least or at most 91, at least or at most 92, at least or at most 93, at least or at most 94, at least or at most 95, at least or at most 96, at least or at most 97, at least or at most 98, at least or at most 99, at least or at most 100. In some embodiments, the overmold layer exhibits a Shore D durometer hardness of about 20-30 or a Shore A durometer hardness of about 20-30.FIG. 1 depicts a non-limiting, exemplary schematic diagram of a ball point flow channel embodiment of a penetrating tip with an overmolded layer 60 of a sample dispensing mechanism (shown in brackets). A flow channel 62 is shown beginning at the distal end of the penetrating tip and running along the length of the penetrating tip. FIG. 6 depicts another non-limiting, exemplary schematic diagram of a ball point flow channel embodiment of a penetrating tip. FIG. 11 depicts a non-limiting, exemplary schematic diagram of a dispensing cap assembly (including a cap 72 and a dispensing rod 74) in which the penetrating tip includes a flow channel 76 and a transition to a larger opening.

[0043] The dispensing rod can include at least one piercing tip. The number of piercing tips can vary. The number of piercing tips can correspond to the number of reaction chambers in the cartridge. For example, a dual reaction chamber cartridge can include a dispensing rod with a dual function piercing tip. The number of piercing tips on the dispensing rod can vary in various embodiments. In some embodiments, the number of piercing tips on a dispensing rod is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 109, 109, 109, 102, 104, 105, 106, 107, 108, 109, 110, 2, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or a number or range between or about any two of these values. In some embodiments, the number of piercing tips on a dispensing rod is min or max 1, min or max 2, min or max 3, min or max 4, min or max 5, min or max 6, min or max 7, min or max 8, min or max 9, min or max 10, min or max 11, min or max 12, min or max 13, min or max 14, min or max 15, min or max 16, min or max 17, min or max 18, min or max 19, min or max 20 at, 21 at min or max, 22 at min or max, 23 at min or max, 24 at min or max, 25 at min or max, 26 at min or max, 27 at min or max, 28 at min or max, 29 at min or max, 30 at min or max, 31 at min or max, 32 at min or max, 33 at min or max, 34 at min or max, 35 at min or max, 36 at min or max, 37 at min or max, 38 at min or max, 39 at min or max, 40 at min or max, 41 at min or max,Min or max 42, Min or max 43, Min or max 44, Min or max 45, Min or max 46, Min or max 47, Min or max 48, Min or max 49, Min or max 50, Min or max 51, Min or max 52, Min or max 53, Min or max 54, Min or max 55, Min or max 56, Min or max 57, Min or max 58, Min or max Maximum 59, Minimum or Maximum 60, Minimum or Maximum 61, Minimum or Maximum 62, Minimum or Maximum 63, Minimum or Maximum 64, Minimum or Maximum 65, Minimum or Maximum 66, Minimum or Maximum 67, Minimum or Maximum 68, Minimum or Maximum 69, Minimum or Maximum 70, Minimum or Maximum 71, Minimum or Maximum 72, Minimum or Maximum 73, Minimum or Maximum 74, Minimum or Maximum 75, Minimum or Maximum 76, Maximum minimum or maximum 77, minimum or maximum 78, minimum or maximum 79, minimum or maximum 80, minimum or maximum 81, minimum or maximum 82, minimum or maximum 83, minimum or maximum 84, minimum or maximum 85, minimum or maximum 86, minimum or maximum 87, minimum or maximum 88, minimum or maximum 89, minimum or maximum 90, minimum or maximum 91, minimum or maximum 92, minimum or maximum 93, minimum or maximum 94, minimum or maximum 95, minimum or maximum 96, minimum or maximum 97, minimum or maximum 98, minimum or maximum 99, minimum or maximum 100, minimum or maximum 200, minimum or maximum 300, minimum or maximum 400, minimum or maximum 500, minimum or maximum 600, minimum or maximum 700, minimum or maximum 800, minimum or maximum 900, or minimum or maximum 1000. The number of reaction chambers coupled to the cartridge body can vary in various embodiments. In some embodiments, the number of reaction chambers coupled to the cartridge body is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or a number or range between or about any two of these values. In some embodiments, the number of reaction chambers coupled to the cartridge body is at least or at most 1, at least or at most 2, at least or at most 3, at least or at most 4, at least or at most 5, at least or at most 6, at least or at most 7, at least or at most 8, at least or at most 9, at least or at most 10, at least or at most 11, at least or at most 12, at least or at most 13, at least or at most 14, at least or at most 15, at least or at most 16, at least or at most 17, at least or at most 18, at least or at most 19, at least or at most 20, at least or at most 21, at least or at most 22, at least or at most 23, at least or at most 24, at least or at most 25, at least or at most 26, at least or at most 27, at least or at most 28, at least or at most 29, at least or at most 30, at least or at most 31, at least or at most 32, at least or at most 33, at least or at most 34, at least or at most 35, at least or at most 36, at least or at most 37, at least or at most 38, at least or at most 39, at least or at most 40, at least or at most 41, at least or at most 42, at least or at most 43, at least or at most 44, at least or at most 45, at least or at most 46, at least or at most 47, at least or at most 48, at least or at most 49, at least or at most 50, at least or at most 51, at least or at most 52, at least or at most 53, at least or at most 54, at least or at most 55, at least or at most 56, 8, min or max 19, min or max 20, min or max 21, min or max 22, min or max 23, min or max 24, min or max 25, min or max 26, min or max 27, min or max 28, min or max 29, min or max 30, min or max 31, min or max 32, min or max 33, min or max 34, min or max 35, min or max 36, min or max 37, min or max 38, min or max 39 for large, 40 for min or max, 41 for min or max, 42 for min or max, 43 for min or max, 44 for min or max, 45 for min or max, 46 for min or max, 47 for min or max, 48 for min or max, 49 for min or max, 50 for min or max, 51 for min or max, 52 for min or max, 53 for min or max, 54 for min or max, 55 for min or max, 56 for min or max, 57 for min or max, 58 for min or max, 59 for min or maximum 60, minimum or maximum 61, minimum or maximum 62, minimum or maximum 63, minimum or maximum 64, minimum or maximum 65, minimum or maximum 66, minimum or maximum 67, minimum or maximum 68, minimum or maximum 69, minimum or maximum 70, minimum or maximum 71, minimum or maximum 72, minimum or maximum 73, minimum or maximum 74, minimum or maximum 75, minimum or maximum 76, minimum or maximum 77, minimum or maximum 78, minimum or maximum 79, minimum or maximum 80,minimum or maximum 81, minimum or maximum 82, minimum or maximum 83, minimum or maximum 84, minimum or maximum 85, minimum or maximum 86, minimum or maximum 87, minimum or maximum 88, minimum or maximum 89, minimum or maximum 90, minimum or maximum 91, minimum or maximum 92, minimum or maximum 93, minimum or maximum 94, minimum or maximum 95, minimum or maximum 96, minimum or maximum 97, minimum or maximum 98, minimum or maximum 99, minimum or maximum 100, minimum or maximum 200, minimum or maximum 300, minimum or maximum 400, minimum or maximum 500, minimum or maximum 600, minimum or maximum 700, minimum or maximum 800, minimum or maximum 900, or minimum or maximum 1000. In some embodiments, the number of seals is min or max 1, min or max 2, min or max 3, min or max 4, min or max 5, min or max 6, min or max 7, min or max 8, min or max 9, min or max 10, min or max 11, min or max 12, min or max 13, min or max 14, min or max 15, min or max 16, min or max 17, min or max 18, min or max 19, min or max 20, min or max 21, min or max 22, min or max 23, min or max 24, min or max 25, min or max 26, min or max 27, min or max 28, min or max 29, min or max 30, min or max 31, min or max 32, min or max 33, min or max 34, min or max 35, min or max 36, min or max 37, min or max 38, min or max 39, min or max 40, min or max 41, min or max 42, min or max 43, min or max 44, min or max 45, min or max 46, min or max 47, min or max 48, min or max 49, min or max 50, min or max 51, min or max 52, min or max 53, min or max 54, min or max 55, min or max 56, min or max 57, min or max 58, min or max 59, min or max 60, min or max 61, min or max 62, min or max 63, min or max 64, min or max 65, min or max 66, min or max 67, min or max 68, min or max 69, min or max 70, min or max 71, min or max 72, min or max 73 34 for maximum, 35 for minimum or maximum, 36 for minimum or maximum, 37 for minimum or maximum, 38 for minimum or maximum, 39 for minimum or maximum, 40 for minimum or maximum, 41 for minimum or maximum, 42 for minimum or maximum, 43 for minimum or maximum, 44 for minimum or maximum, 45 for minimum or maximum, 46 for minimum or maximum, 47 for minimum or maximum, 48 for minimum or maximum, 49 for minimum or maximum, 50 for minimum or maximum, 51 for minimum or maximum, 52 for minimum or maximum, 53 for minimum or maximum, 54 for minimum or maximum, 55 for minimum or maximum, 56 for minimum or maximum, 57 for minimum or maximum, 58 for minimum or maximum, 59 for minimum or maximum, 60 for minimum or maximum, 61 for minimum or maximum, 62 for minimum or maximum, 63 for minimum or maximum, 64 for minimum or maximum, 65 for minimum or maximum, 66 for minimum or maximum, 67 for minimum or maximum, 68 for minimum or maximum,Min or Max 69, Min or Max 70, Min or Max 71, Min or Max 72, Min or Max 73, Min or Max 74, Min or Max 75, Min or Max 76, Min or Max 77, Min or Max 78, Min or Max 79, Min or Max 80, Min or Max 81, Min or Max 82, Min or Max 83, Min or Max 84, Min or Max 85, Min or Max 86, Min or Max 87, Min or Max 88, Min or Max 89, Min or Max The range may be 90, a minimum or maximum of 91, a minimum or maximum of 92, a minimum or maximum of 93, a minimum or maximum of 94, a minimum or maximum of 95, a minimum or maximum of 96, a minimum or maximum of 97, a minimum or maximum of 98, a minimum or maximum of 99, a minimum or maximum of 100, a minimum or maximum of 200, a minimum or maximum of 300, a minimum or maximum of 400, a minimum or maximum of 500, a minimum or maximum of 600, a minimum or maximum of 700, a minimum or maximum of 800, a minimum or maximum of 900, or a minimum or maximum of 1000.

[0044] The volume of the predetermined partial volume of sample fluid can vary in various embodiments. The predetermined partial volume of sample fluid includes at least about 10 μL, about 15 μL, about 20 μL, about 25 μL, about 30 μL, about 35 μL, about 40 μL, about 45 μL, about 50 μL, about 60 μL, about 70 μL, about 80 μL, about 90 μL, about 100 μL, about 110 μL, about 120 μL, about 128 μL, about 130 μL, about 140 μL, about 150 μL, about 160 μL, about 170 μL, about 180 μL, about 190 μL, or about 200 μL, or a value or range between any two of these values.

[0045] Reaction chambers coupled to the same cartridge may contain different reagents selected to perform different respective tests and / or detect different respective target entities. In some embodiments, the number of different types of reagents can be a minimum or maximum of 2, a minimum or maximum of 3, a minimum or maximum of 4, a minimum or maximum of 5, a minimum or maximum of 6, a minimum or maximum of 7, a minimum or maximum of 8, a minimum or maximum of 9, a minimum or maximum of 10, a minimum or maximum of 20, a minimum or maximum of 30, a minimum or maximum of 40, a minimum or maximum of 50, a minimum or maximum of 60, a minimum or maximum of 70, a minimum or maximum of 80, a minimum or maximum of 90, a minimum or maximum of 100, a minimum or maximum of 200, a minimum or maximum of 300, a minimum or maximum of 400, a minimum or maximum of 500, a minimum or maximum of 600, a minimum or maximum of 700, a minimum or maximum of 800, a minimum or maximum of 900, a minimum or maximum of 1000, a minimum or maximum of 10000, or a minimum or maximum of 100000. The reagents can be, for example, lyophilized, heat dried, freeze dried, or in a stable buffer. The test reagents contained in the cartridge prior to testing can be configured for other types of tests that do not necessarily utilize nucleic acid amplification. For example, in some embodiments, direct detection of a chemical reaction can be used to detect the presence of trace elements or additives in the sample. Optionally, immunoassay detection methods can be used to directly conjugate and provide detection of certain proteins in the sample material that is diluted and dispensed into one or more reaction chambers (e.g., test tubes).

[0046] At least one piercing tip that ruptures at least one seal can function to generate one or more flaps. The one or more flaps can include a portion of the at least one seal ruptured by the at least one piercing tip. In some embodiments, the flaps do not adhere to the at least one piercing tip and / or do not disrupt fluid flow through the opening. The piercing tip can include a geometry configured to reduce wicking of sample fluid to the at least one piercing tip and / or the one or more flaps.

[0047] In some embodiments, at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% of the area of ​​the at least one seal is in contact with the at least one penetrating tip. In some embodiments, at least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% of the predetermined sub-volume of the sample fluid, or a value or range between any two of these values, flows into the at least one reaction chamber. In some embodiments, after at least one seal is ruptured, less than about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, or about 1% of a predetermined partial volume of the sample fluid, or a value or range between any two of these values, of the sample fluid remains in or on the dispensing chamber, the at least one seal, and / or the at least one piercing tip.

[0048] 16 depicts a non-limiting exemplary schematic diagram of a dispensing rod including a sagittal penetrating tip having longitudinal grooves 78. At least one penetrating tip can be fluted. At least one penetrating tip can include one or more flow channels. The one or more flow channels can be positioned (i) at a proximal end of the at least one penetrating tip, (ii) at a distal end of the at least one penetrating tip, or (iii) over a length of the at least one penetrating tip. In some embodiments, at least a portion of the predetermined subvolume of the sample fluid flows through the at least one opening through the one or more flow channels. The fluid flow can be at a higher flow rate (e.g., at least about 1.5 times (e.g., 1.5 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times, 100 times, or a multiple or multiple range between any of these values) higher flow rate) compared to a sample inspection system in which the at least one piercing tip does not include the one or more flow channels. The one or more flow channels can include a longitudinal groove extending along the at least one piercing tip. The step of the at least one piercing tip rupturing the at least one seal can include the at least one piercing tip penetrating the at least one seal and moving into at least a portion of the at least one reaction chamber. The at least one opening can expand in size as the at least one piercing tip moves into at least a portion of the at least one reaction chamber. The at least one opening can remain substantially the same size as the at least one piercing tip moves into at least a portion of the at least one reaction chamber. Figure 14 depicts a non-limiting exemplary schematic diagram of a cross-section of a piercing rod provided herein.

[0049] In some embodiments, at least one penetrating tip does not include a sharp tip. FIG. 15 depicts a non-limiting exemplary schematic diagram of a dispensing rod including a penetrating tip without a sharp tip. A distal portion of the at least one penetrating tip can include a flat surface. The flat surface can be angled at about 20°, about 15°, about 10°, about 5°, or less than about 1° with respect to the surface of the at least one seal. In some embodiments, the flat surface of the at least one penetrating tip is angled at about 1°, about 2°, about 3°, about 4°, about 5°, about 6°, about 7°, about 8°, about 9°, about 10°, about 11°, about 12°, about 13°, about 14°, about 15°, about 16°, about 17°, about 18°, about 19°, about 20°, about 21°, about 22°, about 23°, about 24° with respect to the surface of the at least one seal. , about 25°, about 26°, about 27°, about 28°, about 29°, about 30°, about 31°, about 32°, about 33°, about 34°, about 35°, about 36°, about 37°, about 38°, about 39°, about 40°, about 41°, about 42°, about 43°, about 44°, about 45°, about 46°, about 47°, about 48°, about 49°, about 50°, about 51°, about 52°, about 53°, about 54°, about 55°, 56°, 57°, 58°, 59°, 60°, 61°, 62°, 63°, 64°, 65°, 66°, 67°, 68°, 69°, 70°, 71°, 72°, 73°, 74°, 75°, 76°, 77°, 78°, 79°, 80°, 81°, 82°, 83°, 84°, 85°, 86°, The angle can be 87°, about 88°, about 89°, about 90°, about 91°, about 92°, about 93°, about 94°, about 95°, about 96°, about 97°, about 98°, about 99°, about 100°, about 110°, about 120°, about 130°, about 140°, about 150°, about 160°, about 170°, about 180°, or a value or range between any two of these values. In some embodiments, the flat surface of the at least one penetration tip is at an angle of at least 1° min or max, 2° min or max, 3° min or max, 4° min or max, 5° min or max, 6° min or max, 7° min or max, 8° min or max, 9° min or max, 10° min or max, 11° min or max, 12° min or max, 13° min or max, 14° min or max, 15° min or max,min or max 16°, min or max 17°, min or max 18°, min or max 19°, min or max 20°, min or max 21°, min or max 22°, min or max 23°, min or max 24°, min or max 25°, min or max 26°, min or max 27°, min or max 28°, min or max 29°, min or max 30°, min or max 31°, min or max 32°, min or max 33°, min or max 34°, min or max 35°, min or max 36°, min or max 37°, min or max 38°, min or max 39°, min or max 40°, min or max 41°, min or max 42°, min or max 43°, min or max 44°, min or max 45°, min or max 46°, min or max 47°, min or max 48°, min or max 49°, min or max 50°, min or max 51°, min or max 52°, min or max 53°, min or max 54°, min or max 55°, min or max 56°, min or max 57°, min or max 58°, min or max 59°, min or max 60°, min or max is maximum 61°, minimum or maximum 62°, minimum or maximum 63°, minimum or maximum 64°, minimum or maximum 65°, minimum or maximum 66°, minimum or maximum 67°, minimum or maximum 68°, minimum or maximum 69°, minimum or maximum 70°, minimum or maximum 71°, minimum or maximum 72°, minimum or maximum 73°, minimum or maximum 74°, minimum or maximum 75°, minimum or maximum 76°, minimum or maximum 77°, minimum or maximum 78°, minimum or maximum 79°, minimum or maximum 80°, minimum or maximum 81°, minimum or maximum 82°, minimum or maximum 83° , min or max 84°, min or max 85°, min or max 86°, min or max 87°, min or max 88°, min or max 89°, min or max 90°, min or max 91°, min or max 92°, min or max 93°, min or max 94°, min or max 95°, min or max 96°, min or max 97°, min or max 98°, min or max 99°, min or max 100°, min or max 110°, min or max 120°, min or max 130°, min or max 140°, min or max 150°,It may be at least or at most 160°, at least or at most 170°, or at least or at most 180°.

[0050] The at least one reaction chamber may include a trapped gas. The cartridge body may include a gas headspace above the sample fluid. The dispensing rod may be configured to equalize pressure between the gas headspace and the at least one reaction chamber after the at least one seal is ruptured. The at least one penetrating tip may include at least one vent opening leading to a vent lumen extending through the dispensing rod. The dispensing rod may include a vent port positioned in the gas headspace and in fluid communication with the vent lumen of the dispensing rod. The sample dispensing mechanism may include at least one hydrophobic filter. In some embodiments, any fluid passing between the at least one vent port and the at least one vent opening must pass through the hydrophobic filter. The vent opening may be positioned (i) at a proximal end of the at least one penetrating tip, (ii) at a distal end of the at least one penetrating tip, or (iii) over the length of the at least one penetrating tip. The trapped gas displaced by at least one of the penetrating tips and / or the predetermined subvolume of the sample fluid can have the ability to leak into the gas headspace through at least one vent opening. FIG. 7 illustrates a non-limiting, exemplary schematic diagram of a ball point embodiment of a penetrating tip provided herein with a vent opening 64 positioned at the distal end of the penetrating tip. FIG. 8 illustrates a non-limiting, exemplary schematic diagram of a sample dispensing mechanism with a vent opening 66 positioned throughout the length of the penetrating tip. FIG. 9 illustrates a non-limiting, exemplary schematic diagram of a side cross-sectional view of the sample dispensing mechanism shown in FIG. 8 showing the gas vent opening 66 leading to a vent lumen extending through the dispensing rod. FIG. 10 illustrates a non-limiting, exemplary schematic diagram of a penetrating tip with a vent opening 70 positioned throughout the length of the penetrating tip. FIG. 12 illustrates a non-limiting, exemplary schematic diagram of a sample dispensing mechanism with a vent opening 78 positioned at the distal end of the penetrating tip. FIG. 13 depicts a cross-sectional side view, non-limiting exemplary schematic diagram of the sample dispensing mechanism shown in FIG. 12 showing an internal vent path 80 having a hydrophobic filter.

[0051] In some embodiments, a (diagnostic) testing device (also referred to as an "instrument") and a sample testing system (also referred to herein for ease of reference as a "cartridge") suitable for use with the testing meter to perform tests on biological or environmental samples are provided. The cartridges and meters described herein can be easy for a user to operate without the need for common laboratory equipment. A sample testing system including a diagnostic testing assembly and a diagnostic testing device is disclosed in U.S. Patent Application Publication No. 2020 / 0278368, the entire contents of which are incorporated herein by reference.

[0052] In some embodiments, a test cartridge is provided having a removable closure or cap to allow for the addition of a test sample, the cartridge incorporating a cartridge body containing sample preparation fluids such as buffers or lysates to aid in sample preparation and may include isolation of target DNA material from amongst sample cells. The sample preparation fluid reservoir section of the cartridge (e.g., cartridge body) may be a closed volume to securely hold the sample preparation solution until such time as a partial volume (e.g., a pre-determined partial volume of sample fluid from the cartridge body) is dispensed through a perforation in an otherwise well-sealed wall between the reservoir and a reaction chamber coupled thereto. Optionally, the cartridge incorporates chemical and biological reagents required for sample preparation and testing. In some embodiments, these reagents include those configured for nucleic acid amplification using isothermal nucleic acid amplification methods, gene sequence binding, and light output. Optionally, the cartridge incorporates chemical and biological reagents required for sample preparation, nucleic acid amplification, and gene sequence detection using polymerase chain reaction, PCR, and nucleic acid amplification methods.

[0053] In some embodiments, the sample testing system is manufactured prior to the testing (e.g., diagnostic tests) and comes in the form of a disposable diagnostic testing cartridge that is pre-loaded with all of the precursor chemical components (e.g., reagents) to perform a particular set of one or more diagnostic tests. In some embodiments, the sample testing system / cartridge is configured such that it can be safely processed without contamination from the environment, or without causing contamination of the user or the environment with test substances, or without causing interference with these chemical components, or otherwise affecting subsequent actions of the cartridge that may require interaction with a diagnostic testing instrument.

[0054] A user of the sample testing system wishing to perform a test on a biological or environmental sample introduces the sample into the cartridge. At this stage, with the cartridge closure removed, the cartridge provides an unobstructed open volume, i.e., a swab carrying the biological or environmental sample can be conveniently used to agitate the sample preparation fluid within the cartridge body and to flush the biological or environmental sample from the swab into the sample preparation fluid without encountering any obstacles that would impede this stage. However, while this characterizes an open volume within the cartridge, it will be apparent to one skilled in the art that the use of a swab is by no means necessary and that the sample in any suitable form can be added to the sample preparation fluid by any suitable means.

[0055] The sample inspection system may include a closure for sealing the cartridge body and the sample dispensing mechanism therein after receipt of a biological or environmental sample. At least one of the closure and the cartridge body may be configured to prevent or at least inhibit removal of the closure from the cartridge body so that the fluid remains sealed within the sample inspection system. The sample dispensing mechanism may be attached to the closure such that the act of adding the closure to the cartridge body also accomplishes insertion of the sample dispensing mechanism into the cartridge body. In some embodiments, a single action by a user causes the sample dispensing mechanism to rupture at least one seal and dispense sample fluid from the cartridge body into at least one reaction chamber. The single action by a user may be a sustained screwing action applied to the closure against the cartridge body, the screwing action causing actuation of the sample dispensing mechanism to seal the cartridge body. The closure may include a screw thread. The sample inspection system may include a second closure that is removed to seal the sample preparation fluid within the cartridge body prior to use and to allow a biological or environmental sample to be added to the sample preparation fluid contained in the cartridge body.

[0056] In the case of biological samples, the step of adding the sample to the sample preparation fluid in the cartridge body initiates a specific biological or chemical sample dilution and cell lysis process to prepare the sample material, including the contained RNA or DNA nucleic acid, for testing. However, the system is not limited to biological tests and can be used, for example, to detect the presence of trace elements or to measure the amount of these trace elements in any type of sample. Other suitable types of diagnostic tests will be apparent to those skilled in the art in light of the present disclosure.

[0057] The cartridge protects the reagents during shipping and storage prior to running a test and aids in the testing process while a diagnostic test is in progress. In some embodiments, the cartridge retains the test reagents, amplified gene products, and contaminants at all times, including upon completion of the test. The sealed cartridge can be removed for disposal upon completion of the test, and in some embodiments, the meter is protected from fluids and contamination at all times.

[0058] The user can be protected from biological or chemical hazards of the sample during the subsequent testing process after the biological sample has been added to the cartridge and then sealed therein, and after the cartridge has been removed for disposal.

[0059] The systems provided herein (e.g., diagnostic test cartridges) can include one or more reaction chambers, also referred to herein for convenience as "test tubes," intimately coupled to a cartridge body within the cartridge by a separating wall. In some embodiments, the cartridge body is completely sealed from the coupled reaction chambers and is typically supplied pre-filled with a volume of sample preparation fluid and with a removable closure.

[0060] In use, the cartridge can be supported and heated within the testing device and the removable closure removed to add sample, in some embodiments the sample can be any biological or chemical sample incorporated into a test tube to which appropriate diagnostic testing reagents and test-indicating chemistry reagents are coupled.

[0061] In some embodiments, the test cartridge is supplied with an additional cap that has a dispenser mechanism attached. In some embodiments, the additional cap incorporates the dispenser mechanism and is installed after the first cap is removed and sample is added. In some embodiments, when the additional cap and dispenser mechanism are inserted and the cap is closed by an action such as screwing it on, the dispenser mechanism pierces the base of the sample chamber and dispenses a measurement volume of the prepared sample fluid into one or more reaction chambers. The cap is then closed to seal the sample in the cartridge assembly.

[0062] Alternatively, the first cap may have a dispensing mechanism which, when removed, is attached to the first cap in a separate operation to form an additional cap with an associated dispensing mechanism ready to be reattached to perform the dispensing function and close the cartridge.

[0063] Optionally, after the sample is added, the dispensing mechanism is directly inserted on its own, then the cap is fitted and the action of closing this cap, such as screwing the cap closed, activates the dispensing mechanism to close the cartridge.

[0064] In some embodiments, a method of sample testing is provided. In some embodiments, the method of sample testing includes adding a biological or environmental sample into a sample preparation fluid contained in a cartridge body of a sample testing system disclosed herein for preparation of the sample in the system, and after the adding step, inserting a sample dispensing mechanism into the cartridge body and attaching a closure to the cartridge body, and rupturing at least one seal between the cartridge body and the at least one reaction chamber to allow sample fluid to flow from the cartridge body into the at least one reaction chamber, while preventing further fluid transfer between the cartridge body and the at least one reaction chamber, and actuating the sample dispensing mechanism to dispense a predetermined subvolume of the sample fluid from the cartridge body into the at least one reaction chamber for testing in the at least one reaction chamber. The method may include, prior to the adding step, placing the sample testing system into a receiving port of a testing device configured to perform a test on the biological or environmental sample therein.

[0065] Cartridge duplexing Within a single test well, there can be several different markers that will provide light output based on conjugation to several different target gene DNA configurations. In this case, several different sensors or sensors with more than one selective output are used. For example, in a two-channel system, two different fluorescent markers can be used that will be detected by two different fluorescent sensors configured to detect emissions in each frequency range specific to each fluorescent marker to allow the channels to be distinguished.

[0066] A control channel can be provided with the embodiments provided herein, in which the test assay chemistry is configured such that a control target must always be present if the test process is running properly. In this case, the output of the control channel is used to verify that the test process is running properly by the system and to verify that the test results obtained by the other channels and measured by the system are valid. The embodiments provided herein can be used as a multiplex test to test more than one target gene configuration in each test well. Multiple test wells can be used, each processing a differently configured amplification chemistry and a different target marker set. The control channel can operate in one or more wells, and the test can cover the tests performed with the other wells. This configuration allows several tests to be performed on a single sample as different multiplexing approaches.

[0067] Tests (e.g., amplification tests) in a single reaction chamber can be multiplexed in that more than one DNA or RNA target location and control channel can be detected in a single reaction chamber. When the system uses fluorescence as the detection method, different targets can be detected using probes that emit at different fluorescent wavelengths, referred to in the art as detection channels. In the instruments described herein, two detection channels can be included. Using a single tube (e.g., reaction chamber) cartridge described herein and a two-channel detection instrument described herein, the system can allow for detection of two different DNA or RNA targets. In other embodiments, additional reaction chambers can be provided when it is desired to multiplex additional targets from the same sample into a single diagnostic test. In this way, additional target and control channels can be included while using the same number of detection sensors in the instrument. For example, in the case of an embodiment having two instrument sensor channels and two reaction chambers, the system has the capability of detection of four independent DNA or RNA detection channels from a single sample prepared and dispensed from the cartridge body into the reaction chambers. In some embodiments, a cartridge is provided that includes one or more reaction chambers (e.g., test tubes). The number of reaction chambers per cartridge can vary and can be about, minimum or maximum 1, about, minimum or maximum 2, about, minimum or maximum 3, about, minimum or maximum 4, about, minimum or maximum 5, about, minimum or maximum 6, about, minimum or maximum 7, about, minimum or maximum 8, about, minimum or maximum 9, about, minimum or maximum 10, about, minimum or maximum 20, about, minimum or maximum 30, about, minimum or maximum 40, about, minimum or maximum 50, about, minimum or maximum 60, about, minimum or maximum 70, about, minimum or maximum 80, about, minimum or maximum 90, about, minimum or maximum 100, or a value or range between any of these values. The amplification step can include multiplex amplification of two or more target nucleic acid locations.The detection step may include multiplexed detection of two or more nucleic acid amplification products derived from these two or more target nucleic acid configurations. The two or more target nucleic acid configurations may be specific to two or more different organisms. The at least one reaction chamber may be two reaction chambers. The at least one penetrating tip may be two or more penetrating tips. The reaction chamber may include a polymerase chain reaction (PCR) tube. The two reaction chambers may include one or more mixing beads. The reaction chambers may include different reagents selected to perform different tests and / or detect different target entities. The cartridge body may include sample preparation reagents. At least one of the reaction chambers may include one or more reagents suitable for reverse transcription and / or amplification reactions.

[0068] The reaction chamber can include one or more reagents, such as, for example, amplification reagents and nucleic acid detection reagents. The components of the amplification reaction (e.g., one or more amplification reagents) can include, for example, one or more primers (e.g., individual primers, primer pairs, primer sets, oligonucleotides, and multiple primer sets suitable for multiplex amplification), a nucleic acid target (e.g., a target nucleic acid from a sample), one or more polymerases, nucleotides (e.g., dNTPs, and the like), and a suitable buffer (e.g., a buffer containing a detergent, a reducing agent, monovalent ions, and divalent ions). In some embodiments, the amplification can further include a reverse transcriptase and / or a reverse transcription primer. In some embodiments, the amplification reaction can further include one or more detection agents, such as one or more of the detection agents described herein. In some embodiments, the one or more amplification reagents include or consist of a primer, a target nucleic acid, a polymerase, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents comprise or consist of primers, target nucleic acid, polymerase, reverse transcriptase, reverse transcription primer, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents comprise primers, target nucleic acid, polymerase, detection agent, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents comprise or consist of primers, target nucleic acid, polymerase, reverse transcriptase, reverse transcription primer, detection agent, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents consist essentially of primers, target nucleic acid, polymerase, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents consist essentially of primers, target nucleic acid, polymerase, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents consist essentially of primers, target nucleic acid, polymerase, reverse transcriptase, reverse transcription primer, nucleotides, and a suitable buffer. In some embodiments, the one or more amplification reagents consist essentially of primers, target nucleic acid, polymerase, detection agent, nucleotides, and a suitable buffer.In some embodiments, one or more amplification reagents essentially comprise or consist of primers, target nucleic acid, polymerase, reverse transcriptase, reverse transcription primer, detection agent, nucleotides, and a suitable buffer. When one or more amplification reagents essentially consist of certain components, additional components or features may be included that do not have a significant effect on amplification and / or are not necessary to generate detectable products. For example, additional components or features may be included that do not have a significant effect on the function of the components and conditions herein to effect amplification under isothermal conditions to generate detectable amplification products in about 10 minutes or less. Such additional components or features may be referred to as non-essential components, and these components or features may include typical reaction components and / or common additives such as salts, buffers, detergents, ions, oils, proteins, polymers, and the like. In some embodiments, the amplification conditions include an enzymatic activity. Generally, the enzymatic activity is provided by a polymerase, and in some embodiments, by a polymerase and a reverse transcriptase. In some embodiments, the enzymatic activity consists of a polymerase activity. In some embodiments, the enzymatic activity consists of a polymerase activity and a reverse transcriptase activity. Thus, in some embodiments, the enzymatic activity does not include enzymatic activity provided by other enzymes, such as helicases, topoisomerases, ligases, exonucleases, endonucleases, restriction enzymes, nicking enzymes, and recombinases. In some embodiments, the polymerase activity and reverse transcriptase activity are provided by separate enzymes or separate enzyme types (e.g., a polymerase and a reverse transcriptase). In some embodiments, the polymerase activity and reverse transcriptase activity are provided by a single enzyme or enzyme type (e.g., a polymerase).In some embodiments, the amplification comprises one or more of loop-mediated amplification (LAMP), helicase-dependent amplification (HDA), recombinase polymerase amplification (RPA), strand displacement amplification (SDA), nucleic acid alignment-based amplification (NASBA), transcription-mediated amplification (TMA), nicking enzyme amplification reaction (NEAR), rolling circle amplification (RCA), multiple displacement amplification (MDA), ramification (RAM), circular helicase-dependent amplification (cHDA), single primer isothermal amplification (SPIA), signal-mediated amplification of RNA technology (SMART), self-sustained alignment replication (3SR), genomic exponential amplification reaction (GEAR), and isothermal multiple displacement amplification (IMDA).

[0069] In some embodiments, one or more of the amplification reagents can include non-enzymatic components and enzymatic components. Non-enzymatic components can include, for example, primers, nucleotides, buffers, salts, reducing agents, detergents, and ions, and generally do not include proteins (e.g., nucleic acid joining proteins), enzymes, or proteins with enzymatic activity, such as polymerases, reverse transcriptases, helicases, topoisomerases, ligases, exonucleases, endonucleases, restriction enzymes, nicking enzymes, and recombinases. In some embodiments, enzymatic components can include polymerases, or can include polymerases and reverse transcriptases. Thus, such enzymatic components are considered to exclude other proteins (e.g., nucleic acid joining proteins and / or proteins with enzymatic activity), such as helicases, topoisomerases, ligases, exonucleases, endonucleases, restriction enzymes, nicking enzymes, and recombinases.

[0070] The test reagents described herein may further include reagents for detecting and / or quantifying the nucleic acid amplification products. Appropriate detection and quantification reagents can be selected by one of skill in the art based on the detection and / or quantification method selected. The amplification products can be detected and / or quantified by any suitable detection and / or quantification method, including, for example, any of the detection or quantification methods described herein. Non-limiting examples of detection and / or quantification methods include molecular beacons (e.g., real-time, end-point), lateral flow, fluorescence resonance energy transfer (FRET), fluorescence polarization (FP), surface capture, 5'→3' exonuclease activity hydrolysis probes (e.g., TAQMAN), intercalation / conjugation dyes, absorbance methods (e.g., colorimetric, turbidity), electrophoresis (e.g., gel electrophoresis capillary electrophoresis), mass spectrometry, nucleic acid sequencing, digital amplification, primer extension (e.g., iPLEX®), Affymetrix, ELISA ... x molecular inversion probe (MIP) technology, restriction fragment length polymorphism (RFLP analysis), allele specific oligonucleotide (ASO) analysis, methylation specific PCR (MSPCR), pyrosequencing analysis, acycloprime analysis, reverse dot blot, GeneChip microarray, dynamic allele specific hybridization (DASH), peptide nucleic acid (PNA) probes and locked nucleic acid (LNA) probes, AlphaScreen, SNPstream, genetic These include bit analysis (GBA), multiplex minisequencing, SNaPshot, GOOD assay, microarray miniseq, arrayed primer extension (APEX), microarray primer extension, tag arrays, coded microspheres, template-directed incorporation (TDI), colorimetric oligonucleotide ligation assay (OLA), configuration-coded OLA, microarray ligation, ligase chain reaction, padlock probes, invader assays, hybridization with at least one probe, hybridization with at least one fluorescently labeled probe, cloning and sequencing, use of hybridization probes and quantitative real-time polymerase chain reaction (QRT-PCR), nanopore sequencing, chips, and combinations thereof.In some embodiments, detecting the nucleic acid amplification products includes the use of real-time detection methods (i.e., products are detected and / or continuously monitored during the amplification process). In some embodiments, detecting the nucleic acid amplification products includes the use of end-point detection methods (i.e., products are detected after completing or stopping the amplification process). Nucleic acid detection methods can use the use of labeled nucleotides directly incorporated into the target sequence or into a probe containing a complementary sequence to the target. Such labels can be radioactive and / or fluorescent in nature and can be degraded in any of the manners discussed herein. In some embodiments, quantification of the nucleic acid amplification products can be provided using one or more detection methods described below. In some embodiments, the detection methods can be used in conjunction with measurement of signal intensity and / or generation (or reference) of standard curves and / or look-up tables for quantification of the nucleic acid amplification products.

[0071] Figure 17 shows a non-limiting exemplary two-tube cartridge assembly embodiment. While the reaction chambers can be separate test tubes connected to the body of the cartridge, in the example shown in Figure 17, a molded plastic tube component 103 incorporates two internal cavities equivalent to two independent reaction chambers connected to the cartridge body 101. Figure 17 shows the cartridge in a shipping configuration prior to the start of testing, where the shipping cap 102 does not contact the molded latch feature 109 and can be removed by the user at the start of testing.

[0072] The sample preparation reagents may be in liquid form, a portion of which may be added to the reaction chamber to provide an aqueous solution for diluting and exposing the DNA or RNA of the test sample into solution, and may also provide a fluid for dissolving or resuspending lyophilized or dried reagents. The test reagents (e.g., amplification reagents) may be dry or lyophilized or in gel or liquid form for optimal preparation, loading, storage, and shipping.

[0073] In some embodiments, sample preparation fluids and testing reagents (eg, amplification and detection reagents) can be loaded and sealed into the cartridge at the time of manufacture prior to use.

[0074] In a shipping configuration prior to use, the cap can have a shorter length such that its bottom edge does not contact the molded latch cam on the body of the cartridge. This configuration of the shipping cap can allow the cap to seal the sample preparation liquid reagents within the cartridge body, but be removable by the user to initiate a test.

[0075] In some embodiments, the cartridge body has an alignment feature 110 that aligns and engages with a mating slot in the meter to prevent rotation of the cartridge when it is in place in the meter. This anti-rotation feature allows the user to easily remove the shipping cap and then install the inspection cap, all in a one-handed operation.

[0076] The cartridge body includes a seal on its base in such a location that when the cap is attached it forms a sealed container or reservoir that has no fluid communication with the reaction chamber, thereby confining the sample preparation fluid.

[0077] In some embodiments, the reaction chamber is supplied in a separate package and is only clipped or screwed into place onto the cartridge body immediately prior to starting the test.

[0078] A diagnostic test assembly or "cartridge" can include a sample reservoir or chamber, at least one test reservoir or reaction chamber (also referred to herein as an amplification reservoir or chamber), and at least one seal between the sample preparation reservoir and the at least one diagnostic test reservoir to prevent fluid transfer between the sample preparation reservoir and the at least one diagnostic test reservoir. In some embodiments, the sample reservoir or chamber is in the form of a cylindrical cartridge body, and the amplification reservoir or chamber is in the form of an amplification tube coupled to the cartridge body by a locking ring or locking clip and an elastomeric seal. The elastomeric component can provide a seal between the amplification tube and the molded body of the cartridge such that the contents of the amplification tube are not affected by environmental contamination before use and cannot flow out during and after use. In light of the present disclosure, other joining and sealing arrangements and configurations will be apparent to one of skill in the art and may be used in other embodiments. In some embodiments, in a shipping configuration prior to use, the sample reservoir or sample chamber is sealed by a shipping cap and partially filled with a sample preparation fluid or reagent fluid, and the reaction chamber is partially filled with test reagents (e.g., nucleic acid amplification reagents and associated detection probe reagents). These reagents can be in liquid, gel, dry, or lyophilized form. In some embodiments, when a portion of the sample reagent in liquid form is added to the amplification tube, it can be advantageous to provide an aqueous solution for diluting and exposing the DNA or RNA of the test sample into solution, and also to provide a fluid for dissolving or resuspending the lyophilized or dried reagents. The amplification reagents can be dry or lyophilized or in gel or liquid form as is optimal for preparation, loading, storage, and shipping.

[0079] FIG. 18 shows the same cartridge as shown in FIG. 18, where the tube component 103 incorporates two internal reaction chambers 107 and 108. These tubes can hold the required precursor reagents for DNA or RNA amplification and detection probes, typically in dry or lyophilized form. The reagents incorporated in each tube 107, 108 can be different to perform various tests from the same sample, or can be the same reagents to provide additional test reproducibility assurance. The tube component 103 can be clipped into place in a mating recessed feature 104 at the base of the cartridge body 101. An elastomeric seal 105 is housed between the molded cartridge body and the tube assembly, which forms a seal to prevent leakage between the reaction chambers 107, 108 and the environment. Also shown are the female threads 128, at least one seal 321, the cartridge sample volume 106, and the molded latch cam 109.

[0080] The cartridge body contains sample preparation reagents 111, which are typically in liquid form and are typically added during manufacture. It is an option to provide the sample reagents 111 in one or more separate containers and add these reagents to the cartridge when the cap is removed prior to testing. This cartridge can be operated in a similar manner as for the single reaction chamber embodiment, where the sample preparation reagents 111 form an aqueous solution for exposing and holding DNA or RNA from the sample when it is added and for resuspending or dissolving the testing reagents (e.g., amplification reagents) in the tubes 107, 108 when a partial volume (e.g., a predetermined partial volume) of sample diluent fluid is added to the aqueous solution in the tubes 107, 108 by a dispensing action.

[0081] To perform a test, the cartridge can be inserted into the meter port and begin to support and heat the sample reagent fluid 111. This heating assists, accelerates, or enables the sample preparation process, including cell lysis. In some embodiments, the cartridge is supported and operated within the meter, although for illustrative purposes, meter components are not shown in the views of the cartridge shown in Figures 17, 18, 19, 20, 21, 22, 23, 24, and 25.

[0082] FIG. 19 shows the two-tube cartridge at the start of a test with the cap removed and a swab 115 used to add sample material by rinsing the swab 115 in sample reagent fluid 111 contained within the cartridge body 101.

[0083] The sample can be one of many types and can be included in a sample preparation fluid by any suitable method, such as pipetting or dropwise addition of a fluid sample, a small tissue or bodily fluid sample, or addition of an environmental, veterinary, food, or agricultural sample. In some embodiments, the test uses nucleic acid amplification, which can be highly sensitive, and thus the test requires only a small amount of sample material to be effective. The sample or test sample can be any sample isolated or obtained from a subject or a portion thereof. Non-limiting examples of samples include blood or blood products (e.g., serum, plasma, etc.), umbilical cord blood, bone marrow, chorionic villi, amniotic fluid, cerebrospinal fluid, spinal fluid, lavage fluid (e.g., bronchoalveolar lavage fluid, gastric lavage fluid, peritoneal lavage fluid, tube lavage fluid, ear lavage fluid, arthroscopic lavage fluid), biopsy sample, body cavity puncture sample, cells (e.g., blood cells) or portions thereof (e.g., mitochondria, nuclei, or extracts), female reproductive tract washings, urine, feces, sputum, saliva, nasal mucus, prostatic fluid, lavage fluid, semen, lymph, bile, tears, sweat, breast milk, breast fluid, hard tissue (e.g., liver, spleen, kidney, lung, or ovary), the like, or combinations thereof, including fluids or tissues from a subject. The term blood encompasses whole blood, blood products, or any portion of blood, such as serum, plasma, or buffy coat, as defined conventionally. Plasma refers to the portion of whole blood that results from centrifugation of blood that has been treated with an anticoagulant. Serum refers to the watery fluid portion that remains after a blood sample has been coagulated. In many cases, fluid or tissue samples are collected according to standard protocols that are commonly followed by hospitals or clinics. With regard to blood, a suitable amount of peripheral blood (e.g., between 3-40 milliliters) is often collected and can be stored according to standard procedures before or after preparation.

[0084] Suitable samples include, but are not limited to, saliva samples, blood samples, serum samples, plasma samples, urine samples, aspirate samples, and biopsy samples. Thus, the term "sample" with respect to a patient includes blood and other liquid samples from living organisms, solid tissue samples such as biopsy samples, tissue cultures, or cells dispensed therefrom, and their progeny. This definition also includes samples that have been manipulated in any manner after sample preparation, such as by treatment with reagents, washing, or enrichment for certain cell populations, such as cancer cells. This definition also includes samples that have been enriched for a particular type of molecule, such as RNA. The term "sample" includes biological samples such as clinical samples, e.g., blood, plasma, serum, aspirate, cerebrospinal fluid (CSF), and also includes tissue obtained by surgical resection, tissue obtained by biopsy, cultured cells, cell supernatants, cell lysates, tissue samples, organs, and bone marrow. A "biological sample" includes biological fluids dispensed from a living organism (such as, e.g., cancer cells, infected cells), e.g., samples containing RNA obtained from such cells (e.g., cell lysates containing RNA).

[0085] In some embodiments, the source of the sample is a diseased (or suspected) cell, fluid, tissue, or organ. In some embodiments, the source of the sample is a normal (non-diseased) cell, fluid, tissue, or organ. In some embodiments, the source of the sample is a pathogen-infected (or suspected) cell, tissue, or organ. For example, the source of the sample could be an individual, which may or may not be infected, and the sample could be any biological sample taken from the individual (e.g., blood, saliva, biopsy, plasma, serum, bronchoalveolar lavage, saliva, fecal sample, cerebrospinal fluid, fine needle aspirate, swab sample (e.g., buccal swab, cervical swab, nasal swab), interstitial fluid, synovial fluid, nasal secretion, tears, buffy coat, mucosal sample, epithelial cell sample (e.g., epithelial cell scraping)). In some embodiments, the sample is a cell-free liquid sample. In some embodiments, the sample is a liquid sample that may contain cells. Pathogens include viruses, fungi, helminths, protozoa, malarial parasites, Plasmodium parasites, Toxoplasma parasites, Schistosoma parasites, and the like. "Helminths" include roundworms, heartworms, phytophagous nematodes, trematodes, thorny head worms, and cestoda. Protozoal infections include infections from Giardia species, Trichomonas species, African trypanosomiasis, amebic dysentery, babesiosis, balantidial dysentery, Chagas disease, coccidiosis, malaria, and toxoplasmosis. Parasitic / protozoal pathogens include, but are not limited to, Plasmodium falciparum, Plasmodium vivax, Trypanosoma cruzi, and Toxoplasma gondii. Fungal pathogens include, but are not limited to, Cryptococcus and neoformans, Histoplasma and capsulatum, Coccidioides and immitis, Blastomyces and dermatitidis, Chlamydia and trachomatis, and Candida and albicans. Pathogenic viruses include, for example, immunodeficiency viruses (e.g., HIV), influenza viruses, dengue viruses, West Nile viruses, herpes viruses, yellow fever viruses, hepatitis C viruses, hepatitis A viruses, hepatitis B viruses, papilloma viruses, and the like. Pathogenic viruses include papovaviruses (e.g.,Human papillomavirus (HIV), polyomavirus), hepadnavirus (e.g., hepatitis B virus (HBV)), herpesvirus (e.g., herpes simplex virus (HSV), varicella zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), herpes lymphotropic virus, pityriasis rosea, Kaposi's sarcoma-associated herpesvirus), adenovirus (e.g., atadenovirus, avian adenovirus, The viruses may include DNA viruses such as rabies, rabies virus ... Pathogens include, for example, DNA viruses [e.g., papovaviruses (e.g., human papillomavirus (HIV), polyomaviruses), hepadnaviruses (e.g., hepatitis B virus (HBV)), herpes viruses (e.g., herpes simplex virus (HSV), varicella zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), herpes lymphotropic virus, pityriasis rosea, Kaposi's sarcoma-associated herpes virus), adenoviruses (e.g., , atadenoviruses, aviadenoviruses, ictadenoviruses, mastadenoviruses, siadenoviruses), poxviruses (e.g., smallpox, vaccinia virus, cowpox virus, monkeypox virus, orf virus, pseudocowpox, bovine papular stomatitis virus, tanapox virus, yaba monkey tumor virus, molluscum contagiosum virus (MCV)), parvoviruses (e.g., adeno-associated virus (AAV), parvovirus B19, human bocavirus, bufavirus, human parvovirus 4 G1), Geminiviridae, Nanoviridae, and Phycodnaviridae, Mycobacterium tuberculosis, group B streptococcus, methicillin-resistant Staphylococcus aureus, Legionella and pneumophila, group A streptococcus, Escherichia coli, Neisseria gonorrhoeae,Neisseria meningitidis, Streptococcus pneumoniae, Cryptococcus and neoformans, Histoplasma and capsulatum, Haemophilus influenzae type B, Treponema pallidum, Lyme disease spirochete, Pseudomonas aeruginosa, Mycobacterium leprae, Brucella abortus, rabies virus, influenza virus, cytomegalovirus, herpes simplex virus type 1, herpes simplex virus type 2, human serum parvo-like virus, respiratory syncytial virus, varicella-zoster virus, hepatitis B virus, hepatitis C virus, measles virus, adenovirus, human T-cell leukemia virus, Epstein-Barr virus, murine leukemia virus, mumps virus, vesicular stomatitis virus, Sindbis virus, lymphocytic choriomeningitis virus, wart virus, bluetongue virus, Sendai virus, feline leukemia virus, reovirus, poli ... can include mycovirus, simian virus 40, mouse mammary tumor virus, dengue virus, rubella virus, West Nile virus, Plasmodium falciparum, Plasmodium vivax, Toxoplasma gondii, Trypanosoma cruzi, Trypanosoma brucei, Trypanosoma brucei, Schistosoma mansoni, Schistosoma japonicum, Babesia bovis, coccidia of chicken cecum, Onchocerca volvulus, Leishmania tropica, Mycobacterium tuberculosis, Trichinella spiralis, Theileria and parva, Taenia hydatidi, Taenia ovis, Taenia saginata, Echinococcus granulosus, Mesozoon murine, Mycoplasma and arthritidis, Mycoplasma and hydatidi, Mycoplasma and hydatidi, Mycoplasma and arginini, Mycoplasma and laidlowii, Mycoplasma and salivarium, and Mycoplasma and pneumoniae. The pathogen may include one or more of SARS-CoV-2, influenza A, influenza B, and / or influenza C.

[0086] In some embodiments, a sampling swab is introduced into the open cartridge. When a sampling swab is used, the swab is introduced into the sample chamber by the user and rinsed in a sample preparation fluid. The sample preparation fluid can be configured to wash the sample material from the swab and can contain salts, diluent fluids, or detergents that separate cells and cause cell wall lysis to expose the nucleic acid components of the sample material in the sample chamber solution, such that the sample material containing the target material of DNA or RNA is suitable for subsequent nucleic acid amplification.

[0087] Other sampling methods or sample types can be applied to the test cartridge as alternatives to swabs, including, but not limited to, (i) the use of a pipette to add sample fluid, (ii) the use of a drop of whole blood directly from the needle, and (iii) the use of an absorbent pad or membrane to take a fluid sample, such as whole blood, and add it to a sample preparation wash fluid.

[0088] Following the addition of the sample, the instrument display may prompt the user to wait for a period of time to allow sample preparation and cell lysis under the control of the instrument software to take effect. Cell lysis procedures and reagents are known in the art and may generally be performed by chemical lysis techniques (e.g., detergents, hypotonic solutions, and enzymatic procedures, or combinations thereof), physical lysis techniques (e.g., French press and sonication, or electrolytic lysis techniques). Any suitable lysis procedure may be utilized. For example, chemical methods generally involve rupturing cells with a lysis agent, dispensing nucleic acids from the cells, and following these steps with treatment with chaotropic salts. In some embodiments, cell lysis involves the use of detergents (e.g., ionic, nonionic, anionic, zwitterionic). In some embodiments, cell lysis involves the use of ionic detergents (e.g., sodium dodecyl sulfate (SDS), sodium lauryl sulfate (SLS), deoxycholate, cholate, sarkosyl).

[0089] Once the sample preparation period is completed, the user can insert the dispensing cap assembly. Figure 20 shows the cap 120 and dispensing mechanism in position for insertion by the user into the cartridge body 101 in accordance with some embodiments provided herein. The sample inspection system (e.g., dispensing cap assembly) in this figure is comprised of the following visible components: dispensing cap 120, dispensing rod 121, and dispensing chamber (e.g., dispensing insert) 122.

[0090] FIG. 21 shows the dispensing assembly described above in an exploded view. The two-tube cartridge is circular in cross section in the upper section to allow a screw cap to be attached, but has flat sides in the lower section. In some embodiments, the dispensing chamber 122 is a loose sliding fit in the upper circular cross section of the cartridge, but a tight sliding fit in the flat cross section portion of the cartridge. In some embodiments, the tight sliding fit is used to guide the dispensing chamber 122 into a fixed position such that it aligns with a mating feature at the base of the cartridge, and further aligns with the piercing points that allow the two cylindrical bores to allow sample fluid to exit the cartridge body 101 into the two reaction chambers 103. In some embodiments, the transition from the circular cross section to the flat cross section is graduated using a twist of form such that when the dispensing chamber 122 is inserted, the cross section naturally rotates to guide the dispensing chamber into alignment. This transition from round to flat in cartridge body 101 configuration is shown in the body section between lid orientation feature 13 and cap locking feature 109 in FIG.

[0091] A non-limiting exemplary dispensing assembly is shown in an exploded view in Figure 21. In some embodiments, the dispensing rod 121 is clipped into the cap 120 so that it can rotate freely to aid in aligning the dispensing assembly 122 as it enters the flat cross section of the cartridge as it is inserted. In some embodiments, the dispensing rod 121 has two protrusions 305 and 306. The lower section for each of the protrusions 305, 306 forms a corresponding piston with a corresponding O-ring seal 303, 304, and below each piston forms a corresponding piercing tip 307, 308.

[0092] The piercing tips 307, 308 can include a geometry configured to assist fluid flow past the points 307, 308 during penetration. In the described embodiment, the piercing tips 307, 308 can include a geometry configured to generate a large opening in at least one seal. The large opening can include puncturing of at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% of the area of ​​such at least one seal, or a number or range between any two of these values.

[0093] When dispense chamber 122 is assembled into dispense rod 121, piercing tips 307, 308 of dispense rod projections 305, 306 project into the cylindrical bore of dispense chamber 122 but do not fill its volume. These bores are shown in cross section in FIG. 22. In some embodiments, this dispense chamber section is shown in FIGS. 20 and 21 and has a slide 309 that fits closely between and slides over projections 305, 306 of dispense rod 121. Once attached, dispense chamber 122 can slide until it contacts small bridge 310 on the dispense rod. Contact with small bridge 310 prevents further progression during normal handling prior to use, and the sliding attachment of insert 122 onto dispense rod 121 can hold and align dispense chamber 122 in a controlled manner, and is a firm attachment such that normal handling does not result in dispense chamber 122 being dislodged.

[0094] 22 shows a non-limiting exemplary two reaction chamber (e.g., two tube) dispense chamber 122 in cross section partially pressed into the cartridge assembly. In some embodiments, the dispense chamber 122 is slide-mounted onto a rod 121 and is prevented from further progression by a small bridge 310. In some embodiments, the dispense chamber 122 incorporates two open-ended terminated cylindrical bores 301, 302. The dispense rod can include a sealing "O" ring 124.

[0095] 23 shows in cross section where the dispensing mechanism has advanced into the cartridge to the point where the female threads of the screw cap 120 engage the threads on the cartridge body and the dispense chamber 122 has just come into contact with the thin section of material at the base 127 of the cartridge body. In some embodiments, the base of the dispense chamber 122 has features that fit into the base of the cartridge body to form a press fit therewith and form a fluid seal around the base of each of the cylindrical bores 301, 302. In some embodiments, a bridge 310 on the dispense rod 121 allows the dispense rod to apply enough force against the dispense chamber to rigidly seat it into a sealing feature at the base of the cartridge body. In some embodiments, once this portion is seated, further advancement of the cap threads caused by continued rotation of the cap 120 by the user breaks the small plastic bridge 310, allowing the dispense protrusions 305, 306 to proceed further into the dispense chamber 122.

[0096] In some embodiments, at this point the piercing tips 307, 308 begin to pierce the thin section of material at the base of the cartridge body and the piston or syringe features 303, 304 with O-ring seals advance to seal the tops of the two cylindrical bores 301, 302.

[0097] In some embodiments, further travel as the screw cap 120 is further closed by the user causes the protrusion 310 to deflect or break, and the O-rings 303 and 304 on the protrusions 305 and 306 seal the top of the dispensing barrel to form a closed volume of fluid in the respective dispensing bores 301, 302. In some embodiments, when the user completes the cap closing action, the O-ring sealing piston is pushed by the action of the engaged threaded cap 120, forcing the dispensing chamber 122 into the cartridge body 101 and travelling the full distance through the two dispensing insert bores 301, 302 to dispense the captured sample fluid into each of the amplification or testing reservoirs 103.

[0098] In some embodiments, the cartridge body 101 will have 1 to 3 milliliters of sample and sample diluent fluid 111 present, and the dispense action will dispense a small amount of fluid, on the order of about 50 to 100 microliters, into each of the reaction chambers 103. The scale of the portions used can be changed to vary both the sample diluent volume and the volume dispensed into each of the reaction chambers 103 without altering the form of this embodiment.

[0099] 24 shows a non-limiting exemplary cross-sectional view of the cartridge in a fully dispensed configuration. In some embodiments, the dispense cap 120 is longer than the shipping cap, and its bottom edge has an alignment or anti-rotation feature 129 that latches over a molded cam feature 109 on the cartridge body 101. In some embodiments, this latch prevents the cap 120 from being easily removed, ensuring that the test sample is fully sealed within the cartridge after it has been added and the dispense cap is installed. In some embodiments, this locking feature has significant advantages in terms of operator safety and test reliability, preventing contamination of the user and the test system during use and subsequent removal, handling, and disposal of the used cartridge assembly.

[0100] FIG. 25 shows a non-limiting, exemplary overall exterior view of the cartridge in a fully dispensed configuration with the dispensing cap 120 fully positioned and locked onto the cartridge body 101. In some embodiments, the dispensing assembly cap 120 has features in its configuration that aid in rotation and handling, but also includes a unique molded feature 129 that projects outward and / or downward beyond any other features of the cap 120. In some embodiments, the meters disclosed herein can incorporate a sensor that detects the position or proximity of this feature. In some embodiments, the output of this sensor can be used by the meter's controller and its control software to confirm that the cap 120 is fully closed and rotated to the fully closed position. In a typical meter application workflow, the user is prompted to install and close the dispensing assembly cap 120 until such time as the cap full closure feature 129 is detected as described above. In this case, the diagnostic test proceeds to amplify, detect, and generate a test result only after this detection. If the cap 120 is not detected after an extended period of time, the meter can optionally determine that this has caused a fault or misuse and display an error message on the meter's LCD display or communicate to one or more of the data interfaces. This arrangement has the advantage that the test only proceeds to generate a diagnostic result once it has been confirmed that the cartridge has been used properly for an appropriate period of time and the dispense function has been fully completed. This confirmation can allow the meter to self-test, improving the reliability of the final test result.

[0101] In some embodiments, a dispensing assembly is provided having a pressing cap (e.g., a snap-on cap). In some embodiments, the dispensing cap is pressed (e.g., a user applies a force downwards) and snaps into place (on the cartridge body). FIGS. 30A-30D depict non-limiting, exemplary schematic views of the cap assembly and cartridge in an initial position before a snap-on is formed between the snap-on dispensing cap assembly and the dual reaction chamber cartridge. The dispensing cap assembly can be a snap-on dispensing cap assembly 324. The snap-on dispensing cap assembly can include a snap-on dispensing cap 326, a dispensing rod 328, and a dispensing chamber (e.g., a dispensing insert) 330. The cartridge body (e.g., dual reaction chamber cartridge 332) can include a distal end 334 (e.g., an opening). The distal end 334 can include one or more recesses or protrusions (e.g., shelves 336). The snap-on dispensing cap 326 can include one or more snap-on members 338. The snap-on dispensing cap 326 may include a protrusion or tab 340 and may face inward. The snap-on dispensing cap 326 may be configured to snap-fit ​​connection to the distal end 334 (e.g., opening) of the cartridge body. The snap-on dispensing cap 326 may be formed from a material that is softer and / or more flexible than the distal end 334 of the cartridge body, such as a soft rubber or flexible plastic. The snap-on dispensing cap 326 may be formed by injection molding or another suitable molding process known in the art. In some embodiments, the snap-on dispensing cap 326 includes one or more snap-on members 338 configured to engage the distal end 334 of the cartridge. The snap-on members 338 may be configured to form a snap-on engagement with the distal end 334. The snap-fastening member 338 may include one or more protrusions (e.g., projections or tabs 340) extending radially inward, and the distal end 334 may have one or more recesses or protrusions (e.g., shelves 336) that may engage with the one or more recesses or protrusions to form a snap-fastening.In some embodiments, the snap-on member 338 includes one or more projections or tabs 340 configured to engage a portion of the distal end 334 of the cartridge body to complement the snap-on engagement of the snap-on member 338 with the distal end 334. In some embodiments, the distal end 334 includes one or more recesses or projections 336 positioned to engage the projections or tabs 340. For example, the distal end 334 can include a ledge 336 configured to contact the tab 340 and form a suitable connection therewith. The inwardly extending tab 340 of the snap-on dispensing cap 326 can be configured to grip (e.g., form a snap-on engagement therewith) the ledge 336. FIGS. 31A-31D depict non-limiting exemplary schematic views of the snap-on dispensing cap assembly and dual reaction chamber cartridge shown in FIGS. 30A-30D after the snap-on dispensing cap assembly has been fully engaged to form a snap-on. In some embodiments, the snap-on dispensing cap includes an annular ring extending radially inward, the distal end of the cartridge defining one or more recesses or protrusions, the annular ring of the snap-on dispensing cap engaging the one or more recesses or protrusions to form a snap-fit. In some embodiments, the snap-on dispensing cap includes one or more protrusions extending radially inward, the distal end of the cartridge defining one or more recesses or protrusions, the one or more protrusions of the snap-on dispensing cap engaging the one or more recesses or protrusions of the distal end of the cartridge to form a snap-fit. In some embodiments, the closure is snapped onto the cartridge body by a user directly or by a lever coupled to the test instrument. This single action by the user can be applying a downward force to the closure against the cartridge body. The downward force can form a snap-fit ​​between the closure and the cartridge body. In some embodiments, the downward force causes actuation of a sample dispensing mechanism to seal the cartridge body. The downward force can include a downward force of a lever means.The closure may include a snap-on dispensing cap including one or more snap-fitting members configured to form a snap fit with the distal end of the cartridge body upon a single action by a user.

[0102] In some embodiments, the dispensing cap assembly is supplied fully assembled in a protective packet and is removed and inserted by the user, while in other embodiments this is not required. For example, in some embodiments, the sample dispensing mechanism can be attached to a shipping cap that is removed by the user to form a cap assembly, and in some other embodiments, it can be placed into the sample preparation reservoir and coupled to the dispensing mechanism by the act of adding a cap (either the removed shipping cap or a different cap) to the sample preparation reservoir. In some embodiments, the sample dispensing mechanism is operable to break or otherwise rupture or open at least one seal to allow sample fluid to flow from the sample preparation reservoir into the at least one diagnostic test reservoir for diagnostic testing and detection in the at least one diagnostic test reservoir, and to dispense a predetermined sub-volume of sample fluid from the sample preparation reservoir into the at least one diagnostic test reservoir while preventing further fluid transfer between the sample preparation reservoir and the at least one diagnostic test reservoir (e.g., a reaction chamber).

[0103] In some embodiments, a dispensing insert (e.g., a dispensing chamber) is attached to one end of a dispensing rod. In some embodiments, the dispensing rod incorporates a flange that includes a plunger or piston that is inserted into the cylindrical bore or "cylinder" of the dispensing insert. In some embodiments, the piston forms a sliding seal by a close fit with the cylindrical bore, while other embodiments incorporate an elastomeric seal to improve the seal. In some embodiments, an "O" ring is used to improve the seal against the piston as it slides within the cylindrical bore of the dispensing insert.

[0104] In some embodiments, the dispensing insert (e.g., dispensing chamber) incorporates openings in the form of slots in its upper section. In some embodiments, these slots are positioned such that in the initial configuration of the dispensing rod piston when the assembly is inserted into the cartridge, the internal O-ring is positioned above the base of the slot, and furthermore, these slots extend to the outer diameter of the insert so that when the insert is pressed all the way into the sample preparation reservoir, fluid can flow past the outside of, and also through, the cylindrical bore of the insert. In some embodiments, this configuration prevents pressure buildup during insertion and aids in mixing of the sample fluid. Other suitable forms of openings and configurations will be apparent to those skilled in the art, such as holes or grooves, for example, where the insert does not form a seal with the inner wall of the sample preparation reservoir to accomplish this function, and thus allows the insert to easily move through the sample fluid held in the sample preparation reservoir. In some embodiments, the outer diameter and configuration of the dispensing insert allows the insert to be centered and aligned within the sample preparation reservoir when the insert is pressed in, so that the insert also easily drops within the sample preparation reservoir when inserted (without encountering significant resistance from the sample fluid). In some embodiments, this allows the base of the dispensing insert to be precisely aligned with a mating recess in the base of the sample preparation reservoir.

[0105] In some embodiments, the dispensing rod includes a piston flange with a sealing "O" ring and further includes a piercing tip at the end of the dispensing rod. In some embodiments, when the dispensing assembly is fully inserted, female threads in the cap engage male threads on the body of the cartridge. In some embodiments, when these threads engage one another, the user is prompted and able to gradually screw the cap shut. In some embodiments, the action of screwing the cap shut engages an internal component and pierces a seal at the base of the sample preparation reservoir, providing a mechanical advantage that facilitates progression of the dispensing assembly through the sample preparation reservoir to dispense a sample sub-volume of sample fluid from the sample preparation reservoir into the diagnostic test reservoir.

[0106] In some embodiments, the dispense insert (e.g., dispense chamber) is held on the dispense rod such that after contacting the base of the cartridge body, some additional force is required before the dispense rod can enter deeper into the bore of the dispense insert. In some embodiments, this additional force allows the base of the dispense insert to press under friction or snap into place within a mating feature or surface feature in a recess in the base of the sample preparation reservoir to form a fluid seal with the sample preparation reservoir. In some embodiments, a small elastomeric seal is included on either the dispense insert or the sample tube to assist in forming this seal. However, in some embodiments, the injection molded form base of the dispense insert and the mating feature in the sample preparation reservoir are sufficient to form a fluid seal under compressive forces when these parts are in contact with each other. In some embodiments, there is a detent formed by a circular groove in the dispense rod and a corresponding annular ring on the dispense insert. In some embodiments, the detent provides an initial breakaway force that attempts to lock and seal the dispensing insert in place within the base of the sample preparation reservoir when the resistive force of the detent is overcome under continued rotational action of the screw cap and the dispensing rod begins to advance through the dispensing insert prior to completion of the dispensing operation. Other arrangements for providing a dispensing insert sealing force are available and will be apparent to those of skill in the art in light of the present disclosure.

[0107] In some embodiments, the insert is mounted onto the dispense rod by capturing a collapsible or crushable spacer between it and an engagement feature extending from the dispense rod. In some embodiments, the dispense insert contacts the base of the sample chamber of the cartridge when the dispense cap assembly is pushed into the cartridge by the screw cap action. In some embodiments, the collapsible spacer allows the screw action to apply the force involved in the sealing action, and the base of the cylindrical bore of the dispense insert is forced into a mating feature in the base of the cartridge. In some embodiments, when the cap screw action applies additional force and advancement, the spacer is configured to crush in a controlled manner to push the dispense insert into place and then allow the "O" ring plunger on the dispense rod to enter the tubular bore section of the dispense mechanism. In some embodiments, after the dispense insert is held or locked in place, the "O" ring plunger on the dispense rod enters the tubular section of the dispense mechanism to form a piston and cylinder or syringe. In some embodiments, as the "O" ring plunger captures the fluid volume into the dispense tube, the piercing tip of the dispense rod pierces a plastic section in the cartridge at the base of the tube in the dispense insert. In some embodiments, this piercing action punches a hole through this plastic section and also through the foil or plastic membrane covering the top of the amplification tube. In some embodiments, continued progression of the plunger then dispenses the captured fluid volume into the amplification tube. In some embodiments, the fluid captured in this cylindrical section is a fixed, pre-determined volume of sample fluid that is dispensed through a perforation in the base of the sample chamber into the amplification tube mounted below it.

[0108] Optional chamber fluid dispensing function In some embodiments, when a dispensing chamber (e.g., a dispensing insert) is inserted into the cartridge body 1, sample fluid flows around the dispensing chamber and through the open-ended cylindrical dispensing bore, as shown in FIG. 26. FIG. 27 shows the dispensing chamber in an isometric view. In some embodiments, the dispensing chamber 22 has fins with flow channels that run past the outside of the cylindrical bore, as well as slots that run down only a portion of the length of the dispensing chamber. As a result, sample fluid that flows into the cylindrical bore can flow out through the slots when the sealing plunger is not fully pressed into the solid or "non-slotted" portion of the dispensing chamber 22. FIG. 26 shows typical fluid flow lines. This description can be equally applied to the case of a dispensing chamber in a multiple reaction chamber cartridge, such as the two-tube insert 122 shown in FIG. 21.

[0109] Filter Components In some embodiments, the dispensing chamber is initially configured such that when the sample dispensing mechanism is inserted into the cartridge body, the sample fluid is forced to flow around the outside of the dispensing chamber before it can flow into the dispensing chamber, and the fluid flowing around the outside of the dispensing chamber is forced to flow through a filter or porous filling material that incorporates biological or chemical components that retain and / or capture particles and contaminants, and / or bind to or capture sample fluid components that may otherwise inhibit or interfere with sample testing.

[0110] FIG. 28 shows an alternative embodiment of the dispense chamber (e.g., dispense insert) 22 that allows the inclusion of a filter 322 to remove particles or inclusions therein from the sample fluid that flows into the dispense chamber and is subsequently dispensed into the coupled reaction chamber or reaction tube. In this embodiment, the dispense chamber 22 has its unwanted inlet to a cylindrical bore 320 closed by a seal (e.g., membrane) 321 on its underside. When the dispense chamber 22 is pushed into the cartridge assembly after the sample is added, all of the pushed out sample fluid flows through flow channels provided around the outside of the closed cylindrical bore 320. As shown by the cross-hatching in FIG. 28, one or more filter components 322 can be included in these flow channels. These filters 322 can be fiber-based materials such as compressed glass fibers or porous foam or porous plastic materials. The one or more filter components 322 can be a single annular material disk or several smaller sections placed in each of the available flow paths.

[0111] In some embodiments, the filter component 322 physically captures or traps particles or substances that would otherwise contaminate the sample fluid to be dispensed into the testing reservoir. The filter component 322 can incorporate biological or chemical components that bind to or capture components of the sample fluid that may otherwise inhibit or interfere with the testing or amplification process. In some embodiments, the fluid through the filter component 322 will then fill the central cylindrical dispensing bore 320 from the top through the dispenser slots as portions of these slots are submerged in the sample fluid. This filtered sample fluid can then be made available in the dispenser bore 320 for subsequent sealing and dispensing through a pierce into an attached test tube.

[0112] Magnetic bead-based nucleic acid enrichment In some embodiments, the cartridge body contains one or more magnetic particles along with a sample preparation fluid, the surfaces of which are coated or functionalized to bind to and capture at least one predetermined target chemical species of a biological or environmental sample when mixed into the sample fluid, the sample dispensing mechanism is configured to force the sample fluid through the dispensing chamber when the sample dispensing mechanism is inserted into the cartridge body, one or more magnets are positioned in close proximity to an inner surface of the dispensing chamber such that magnetic particles contained within the sample fluid and that have captured the target chemical species are attracted to and held against the inner surface of the dispensing chamber, and a plunger mechanism that forms a sliding seal with the inner surface of the dispensing chamber thereby collects the magnetic particles held against the inner surface and dispenses them into at least one reaction chamber, generating an increase in the concentration of at least one predetermined target chemical species in a predetermined subvolume of the sample fluid dispensed therein.

[0113] FIG. 29 shows an alternative configuration of the dispensing chamber (e.g., dispensing insert) 22 that allows the magnetic bead enrichment function to occur within the cartridge assembly. In this embodiment, the dispensing chamber 22 does not have any flow channels passing outside the cylindrical bore 320, and all of the sample fluid pushed out when the dispensing chamber 22 is inserted into the cartridge flows through the cylindrical bore 320. In this embodiment, the sample preparation fluid contains magnetic particles or these particles can be added as an examination process step. In some embodiments, the surface of the particles is coated or functionalized to bind and capture at least one target species of interest that is mixed or dissolved in the sample fluid. For example, nucleic acid material, DNA material, or RNA material are mixed in the sample fluid contained in the sample volume 6 of the cartridge 1, so a typical application is to bind these particles onto the functionalized surface coating of the magnetic particles. The magnetic particles can be very small, typically within 0.5 micrometers to 10 micrometers. These magnetic particles remain freely mixed and suspended within the sample fluid, binding to target molecules and capturing them on their own surface coating.

[0114] In some embodiments, the dispense insert or dispense chamber is configured to have a sliding seal with the inner surface of the cartridge, where the solid section of the insert 322 blocks fluid flowing past the outside of the central cylinder, thereby forcing all of the fluid in the sample chamber to flow through the central cylindrical bore 320. The flow lines in FIG. 29 show a typical flow path. In this embodiment, the dispense chamber component incorporates one or more permanent magnets 331 that are encased in the molded plastic and positioned near the inner surface of the cylindrical bore 320. A typical arrangement is to use a ring magnet 331 that surrounds the inside of the bore 320, where the magnet 331 is introduced into the molding process when the insert 322 is injection molded and is encased in the plastic structure of the part 322. In some embodiments, as sample fluid flows through the dispensing cylinder in proximity to the internal magnet 231, the magnetic particles are attracted by the magnetic field against the side wall of the dispensing cylinder and are retained along with any DNA or RNA material captured within the cylindrical dispensing tube 320.

[0115] The piercing component breaks through the thin material at the base of the sample chamber 1 as the dispensing component is forced into the seal at the base of the sample chamber and the piston component engages and seals the top of the chamber. During the dispensing process, the magnetic beads magnetically held against the inner wall of the cylinder are swiped down the bore 320 by the O-ring sealing plunger, thereby mixing back into the sample fluid captured in the dispensing cylinder, and all of this fluid and magnetic beads are dispensed into the coupled test tube by the progressive advancement of the piston into the reaction chamber. This concentrates DNA or RNA material in the sample fluid and delivers it into the reaction chamber 107, 108. This has the advantage of concentrating and purifying the DNA or RNA nucleic acid material removed from the sample, resulting in a more sensitive and reliable diagnostic test. Reagents in the test tube 107, 108 can react with the molecules selectively bound to the magnetic particles as they are eluted by additional sample fluid. The test tube reagents may contain salts, chemicals, or pH suitable for the release of captured substances from the surfaces of the magnetic particles in the reaction chambers 107, 108 and aid in the reaction and detection of these components.

[0116] Manual operation Visual reading Non-instrumented Cartridge operation In some applications, the cartridge is used manually without a meter. For example, in some embodiments, the cartridge is held in one hand, the first cap is removed with the other hand, sample is added, and a second (dispensing) cap is put on and screwed shut. In some such embodiments where the reaction chamber is visually transparent, the dispensing of fluid into the reaction chamber can be visually observed, and a change in color or turbidity over time can be observed to allow a reading or indication of the diagnostic test. This approach takes advantage of operating with a cartridge that is completely sealed once sample is added, and of dispensing a measured volume of diluted and prepared sample fluid internally into the test tube without the use of an external fluid transfer step.

[0117] Optionally, a simple stand can be provided to support the cartridge for the purposes of removing the first cap, adding sample, and fitting and closing the dispensing cap and corresponding mechanism.

[0118] Optionally, a heater block can be provided to provide temperature control of the sample and test tube chambers of the cartridge assembly, while the cartridge is manually withdrawn to observe the test results visible in one or more coupled reaction chambers.

[0119] Inspection Instruments In some embodiments provided herein, the cartridge can be operated in a testing device or "meter" to perform a test (e.g., a diagnostic test). The sample testing system disclosed herein can include a testing device / meter. Below, cartridges and testing meters according to some embodiments of the configurations, systems, and methods provided herein are described. By preloading sample preparation and testing reagents into the cartridge, the sample testing system can be configured to perform a specific, pre-defined set of one or more tests (e.g., diagnostic tests) and provide at least one indication of the test results. Different versions of the cartridges can be manufactured with the same physical configuration but different loaded reagents to cover a wide range of test types and diagnostic applications. In some embodiments, the meter can automatically determine the type of diagnostic test to be performed from the cartridge identifier (visual or otherwise), perform the determined diagnostic test, and provide the diagnostic test results to the user by displaying the result / s on a user interface display upon completion of the diagnostic test and / or by providing the result / s in the form of one or more electronic records or other electronic data through any of several communication interfaces of the meter.

[0120] In some embodiments, a testing device is provided. The testing device may include a receiving port configured to receive a sample testing system provided herein. The testing device may be configured to perform a test on a biological or environmental sample therein. In some embodiments, the testing device further includes a lever means configured to apply a downward force to a sample testing system positioned in the receiving port. The single action may be to apply a downward force to the closure against the cartridge body through the lever means. The downward force may form a snap-fit ​​between the closure and the cartridge body. In some embodiments, a meter (e.g., testing device) is provided having a lever means (e.g., a hinged lid). The hinged lid on the meter may allow a user to depress the closure (e.g., a dispensing cap) with less force due to mechanical advantage. In some embodiments, the lid is stored vertically within the sleeve. In some embodiments, during operation, a user lifts the hinged lid and pivots it to a horizontal position. 32A-32B show non-limiting, exemplary schematic diagrams of the test device with the snap-on dispensing cap assembly and dual reaction chamber cartridge (in an initial position before a snap-fit ​​is formed therebetween) positioned in the receiving port, whereas FIGS. 33A-33B depict non-limiting, exemplary schematic diagrams of the test device shown in FIGS. 32A-32B after the hinged lid is pressed over the top of the snap-on dispensing cap to form a snap-fit. The test device 346 can include a hinged lid 342. The test device 346 can include a sleeve 344 for storing the hinged lid 342 when not in use. The hinged lid 342 can include an increased or raised portion (e.g., ridge 348) configured to contact a surface of the closure. The test device can include a sleeve for storing a lever means. The hinged lid can be substantially parallel to the cartridge body when stored in the sleeve.In some embodiments, at least a portion of the hinge-lid can be configured to slide up and out of the sleeve when lifted by a user to expose a hinge of the hinge-lid, In some embodiments, when the hinge is exposed, the hinge-lid can be capable of pivoting to a horizontal position substantially perpendicular to the cartridge body.

[0121] The testing device can include one or more mating slots configured to align with and engage with one or more alignment features of the cartridge body. The one or more mating slots can be located in the receiving port. The one or more alignment features can prevent rotation of the cartridge body when the cartridge body is in place in the testing device. The one or more alignment features can enable a user to remove the second closure with a one-handed operation and / or perform a single action.

[0122] The cartridge can be supported by, aligned with, heated and / or measured by a sample testing device / meter to assist in closure removal, warm-up, sample addition, sample preparation, sample dispensing, cartridge closure and test result measurement. In some embodiments, the meter includes separate heater areas for independent temperature control of the sample preparation and reaction chambers in the cartridge. In some embodiments of a test sequence, a cartridge containing sample preparation fluid is inserted into the meter, which detects the presence of the cartridge and begins to warm the sample preparation fluid. When the sample preparation fluid reaches a desired temperature, the meter can then prompt the user to add the biological or environmental sample to be analyzed. Heating the sample preparation fluid can be advantageous in assisting in fast and efficient sample preparation.

[0123] Thereafter, or when prompted by the meter, the user can then add a closure to the cartridge body, and this action of actuating the closure not only seals the sample and sample preparation fluids within the cartridge, but can also actuate a dispensing mechanism within the cartridge body to deliver a predetermined volume of a sub-sample into one or more reaction chambers within the cartridge.

[0124] In some embodiments, the instrument then controls the temperature of one or more reaction chambers and the temperature of the sample fluid and test reagents contained therein. This temperature control can maintain a fixed temperature, or can follow a predetermined time-varying temperature profile, for example, or can be subjected to thermal cycling by heating and cooling between various fixed temperatures in the case of a PCR reaction. In either case, a cycle or time series of optical measurements of the contents of the reaction chambers can be taken by the instrument. The instrument can process these measurements to determine a test result that can then be displayed to a user or otherwise provided as an output.

[0125] Provided herein is (i) a meter housing having an access port for receiving a plastic cartridge assembly; (ii) a sensor or switch for detecting the insertion or presence of a cartridge inserted into the device; (iii) controller electronics and associated internal electronics, microprocessor and memory for executing software programs and storing data for future recall and use; (iv) peripheral interfaces for USB, serial or Ethernet connectivity and electrical mating connectors for connection to external memory devices; (v) embedded software for providing the functionality to sequence the processing of the meter, cartridges and obtain diagnostic test measurements for determining test result interpretations; (vi) Configurations, systems and methods comprising and / or using a test instrument or device including one or more of: (i) a temperature-controlled sample chamber heater block for providing heating and temperature control of an upper sample chamber section of a cartridge assembly; (ii) a temperature-controlled heater block for providing heating and temperature of an upper contact-specific reaction chamber (e.g., an amplification test well) within an inserted cartridge and capable of applying controlled temperature, including temperature cycling, to fluids within the cartridge wells; and / or (viii) sensors for detecting and providing measurements of optical absorption, fluorescence, or bioluminescence characteristics of the reaction of reagents and additional sample fluids within the reaction chamber during and upon completion of a test.

[0126] The instrument device may incorporate one or more optical sensors, where these sensors may be scanned along a row of test wells, allowing multiple measurements to be recorded for each test well using one or more different sensors. In some embodiments, the instrument controller may be located remotely from the physical body, for example on a remote server, and the instrument controller may manage and control the operation of the device over a communication network such as the Internet. In some embodiments, one or more of the sensors are coaxial fluorescent sensors, where optically filtered emission light from a light emitting diode or laser illumination light of a selective wavelength range is emitted from the sensor lens. In some embodiments, this illumination light provides optical excitation of the sample in the test well, and this same lens further captures fluorescent emission light of different shifted wavelengths from the sample. In some embodiments, this sample fluorescent emission light is measured and forms a measurement used to determine the diagnostic test result. In some embodiments, one or more of the sensors may detect fluorescence in the sample contained in each test well, using a separate excitation illumination source to optically excite the test sample and a separate sensor to measure the resulting fluorescent emission light. In some embodiments, one or more of the sensors measure optical reflection or optical absorption in the test sample contained in each test well using reflectance or transmittance of a particular optical illumination wavelength range. In some embodiments, one or more of the sensors measure optical emission from the test sample, where the emission is caused by luminescence or bioluminescence in the test sample. In some embodiments, the sensor is scanned past all of the wells at a constant speed, resulting in a large number of measurements. Subsequent processing of this measurement data set can determine which measurements to assign to each test well. This analysis can take into account characteristics such as the relative position or time of acquisition of each measurement, and local peaks associated with an interpolated curve that encompasses the acquired measurements.

[0127] In some embodiments, the instrument device incorporates one or more ultraviolet light sources, where the ultraviolet illumination light can be turned on or off by the instrument controller. In some embodiments, the instrument device incorporates one or more reference targets in the field of view of the fluorescent or optical absorption sensor. In some embodiments, the inspection device includes at least one sensing component configured to determine the degree of closure rotation and / or thread advancement, and is configured to prompt a user to complete the closure actuation if the at least one sensing component determines that the closure actuation is incomplete, and to automatically proceed to the next stage of the diagnostic test if the closure actuation is determined to be complete.

[0128] In some embodiments, at least one of the at least one reaction chambers is transparent, and the testing device is configured to determine a test result in the at least one reaction chamber by detecting or measuring a change in emission and / or absorption at one or more wavelengths in the at least one reaction chamber, and optionally configured to illuminate the at least one reaction chamber to improve or provide the detection or measurement. In some embodiments, the testing device and sample testing system (e.g., diagnostic testing assembly) include respective alignment and support features configured to interengage with each other to ensure that the sample testing system is received in a predetermined alignment with the testing device and to maintain the alignment when a closure is added to the cartridge body and sample dispensing mechanism therein following receipt of a biological or environmental sample.

[0129] The testing device may include one or more components configured to apply a changing and / or moving magnetic field to the sample testing system to effect corresponding movement of magnetic particles within at least one of the cartridge body and the at least one reaction chamber, thereby causing mixing of the sample and sample preparation fluid within the cartridge body and the at least one reaction chamber.

[0130] In some embodiments, the testing device and sample testing system are configured to allow the testing device to independently control the temperature of the cartridge body and at least one reaction chamber.

[0131] In some embodiments, the inspection device includes one or more image sensors configured to generate image data representative of one or more images of at least a portion of the sample inspection system, the images depicting at least one of: (i) a fluid distribution in at least one of the at least one reaction chamber and the cartridge body, where the inspection device is configured to process the image data to monitor the dispensing of sample fluid and proceed to a next stage of the diagnostic test when the monitoring stage determines that dispensing is complete; and (ii) a volume of fluid contained in the at least one reaction chamber, where the inspection device is configured to process the image data to enable compensation for volume tolerances of the dispensed fluid and to enable improved test result determination. In some embodiments, the inspection device includes one or more optical sensors mounted on a translation stage under the control of a controller of the inspection device such that the optical sensors can measure optical absorption, optical emission, or fluorescence from one or more selected reaction chambers of the sample inspection system.

[0132] In some embodiments, the testing device includes at least one ultraviolet (UV) emitting source for denaturing a sample contained within the sample testing system following a diagnostic test to inhibit contamination if the sample fluid flows out of the sample testing system.

[0133] One or more image sensors, which may be incorporated into the meter, may capture digital images of the cartridge, the advancement of the dispense mechanism components, and the state and progression of the fluid contained within the cartridge. The image data obtained by the image sensor and subsequent image analysis may be used by the controller to determine the level of the dispensed sample fluid in each of the reaction chambers 107, 108, which may be used to determine proper completion of the sample fluid dispense. The level of the fluid dispensed in each of the one or more reaction chambers 107, 108 in the cartridge may be used to compensate the test results for tolerances in the dispense operation. The level of the fluid associated with each test tube may be converted to a volume by the controller using a mathematical model of the tubes 107, 108 or by using a look-up table. The volume of the dispensed fluid may affect the concentration of the test reagent in the test chamber fluid after elution into the dispensed fluid. By measuring the volume of the dispensed sample fluid, the concentration of the reagent in each test tube 107, 108 may be calculated. Knowing the effect of test reagent concentration on the test result and on the interpretation of the test time series measurements to interpret it, from a series of previous experiments or from a model of the test response, can be adjusted or compensated for in the device. The fluid sample preparation reagent stored in the cartridge can be colored with a dye. The dye can be used by an image sensor to visually image the colored or contrasted fluid flow into the cartridge reaction chambers 107, 108 to confirm the dispense action and further to confirm the dispense volume. Image analysis of the image of the fluid dispensed into one or more of the coupled reaction chambers 107, 108 can be used to measure the volume in the tubes 107, 108, and this measurement can be used to compensate the test result calculation for the amplification volume. This compensation can be particularly significant for quantitative test results where the concentration of the reagent in the test tubes 107, 108 can affect the measurement and reaction response.

[0134] Although some embodiments use optical measurements of the reaction chamber to determine the test result, sensors using alternative measurement methods can be operated in the same testing device / instrument with the diagnostic test cartridges described herein. These sensors can use magnetic, electrical, atomic, or physical properties of the test fluid to obtain measurements appropriate to determine the test result.

[0135] Cartridge body and reaction chamber mixing Some embodiments of the configurations and methods provided herein contemplate mixing of the contents of either the cartridge body or the reaction chamber, which in some embodiments can improve the reliability or accuracy of the test. Magnetic inserts, such as small steel or ferrite pellets, can be included in the cartridge body and / or reaction chambers 107, 108 to induce mixing during reagent loading of the cartridge (e.g., during initial manufacture of the cartridge). Some embodiments contemplate applying an external magnetic field to the cartridge body (e.g., provided by the testing device moving a permanent magnet or multiple magnets into the vicinity) to induce mixing in the sample fluid and / or reaction chamber. Mixing in the cartridge body can be used to mix the introduced sample material with sample preparation fluid to prepare the sample material for amplification. This preparation mixing can improve cell lysis in the sample, as well as extraction and preparation of target DNA or RNA nucleic acid material.

[0136] In at least some of the above-described embodiments, one or more elements used in an embodiment may be used interchangeably in another embodiment, unless such an exchange is technically feasible. It will be understood by those skilled in the art that various other omissions, additions, and modifications may be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and variations are intended to fall within the subject matter defined by the claims.

[0137] With respect to the use of substantially any plural and / or singular terminology herein, one of ordinary skill in the art can convert such terms from plural to singular and / or from singular to plural where appropriate with respect to relevance and / or application. Various singular / plural permutations may be expressly set forth herein for purposes of clarity. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Any reference to "or" herein is intended to include "and / or" unless otherwise specified.

[0138] In general, it will be understood by those of skill in the art that terms used herein, particularly in the claims (e.g., the body of the claims), are generally intended to be "open" terms (e.g., the term "including" should be interpreted as "including, but not limited to," the term "having" should be interpreted as "having at least," and the term "including" should be interpreted as "including, but not limited to"). Where a particular number of introductory claims is intended, such intent shall be expressly set forth in the claim, and in the absence of such recitation, it will be further understood by those of skill in the art that no such intent exists. For example, as an aid to understanding, the claims below may include the use of the introductory phrases "at least one" and "one or more" to introduce claim subject matter. However, the use of such phrases should not be construed as suggesting that the introduction of a claim recitation without such a phrase limits any particular claim, including the claim recitation so introduced, to embodiments including only one such recitation, even if the same claim includes a claim recitation with the introductory phrase "one or more" or "at least one" and a claim recitation without such an introductory phrase, and even to claims including a claim recitation with the indefinite article "a" or "an." Moreover, even if a specific number of introduced claim recitations is explicitly recited, it will be recognized by those skilled in the art that such recitation is interpreted to mean at least the recited number of introduced claim recitations (e.g., the simple recitation "two recitations" without other modifiers means at least two recitations or two or more recitations).Moreover, in instances where idiomatic expressions similar to "at least one of A, B, and C" are used, such syntax is generally intended to be the meaning that one of ordinary skill in the art would understand the idiomatic expression (e.g., "a system having at least one of A, B, and C" includes, but is not limited to, systems having A only, B only, C only, A and B together, A and C together, B and C together, and / or A, B and C together). It will be further understood by those of ordinary skill in the art that virtually any disjunction word and / or disjunction phrase providing two or more alternative items, whether in the specification, claims, or drawings, must be understood to contemplate the possibility of including one of the items, either of the items, or both items.

[0139] In addition, when features or aspects of the present disclosure are described in terms of a Marker-Mail group, it will be recognized by those skilled in the art that the present disclosure is also thereby described in terms of any individual member of the Marker-Mail group or subgroups thereof.

[0140] As will be understood by those skilled in the art, all ranges disclosed herein, for any and all purposes, including providing a written specification, will encompass any and all possible subranges and combinations of subranges of the ranges. Any ranges recited will be readily recognized as fully descriptive and allowing such ranges to be broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, a middle third, and an upper third. As will also be understood by those skilled in the art, all phrases such as "up to," "minimum," or "at least," "greater than," "less than," etc., include the recited numbers and subsequently refer to ranges that can be broken down into subranges as discussed above. Finally, as will be understood by those skilled in the art, a range includes each individual component. Thus, for example, a group having 1 to 3 items means a group having 1, 2, or 3 items. Similarly, a group having 1 to 5 items means groups having 1, 2, 3, 4, or 5 items, and so forth.

[0141] While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those of ordinary skill in the art. The various aspects and embodiments disclosed herein are intended to be illustrative and not limiting, with the true scope and spirit being indicated in the following claims.

Claims

1. 1. A sample inspection system, comprising: a cartridge body for receiving a biological or environmental sample into a sample preparation fluid contained in the cartridge body to prepare a sample fluid; at least one reaction chamber coupled to the cartridge body; at least one seal that prevents fluid transfer between the cartridge body and the at least one reaction chamber, the at least one seal being located between the cartridge body and the at least one reaction chamber, the at least one seal being capable of being pierced by a piercing tip, thereby allowing fluid transfer between the cartridge body and the at least one reaction chamber; a sample dispensing mechanism for insertion into the cartridge body; Including, the sample dispensing mechanism is operable to rupture the at least one seal to allow sample fluid to flow from the cartridge body into the at least one reaction chamber and to dispense a predetermined subvolume of the sample fluid from the cartridge body into the at least one reaction chamber for testing therein while preventing further fluid movement between the cartridge body and the at least one reaction chamber; the sample dispensing mechanism includes a dispensing rod including at least one piercing tip that ruptures the at least one seal by forming at least one opening in the at least one seal, the at least one piercing tip including a geometry configured to generate a large opening in the at least one seal; Sample inspection system.

2. a closure for sealing the cartridge body and the sample dispensing mechanism therein after receiving the biological or environmental sample; at least one of the closure and the cartridge body is configured to prevent or at least inhibit removal of the closure from the cartridge body so that the fluid remains sealed within the sample testing system; The sample inspection system of claim 1 .

3. The sample testing system of claim 2 , wherein the sample dispensing mechanism is attached to the closure such that the act of adding the closure to the cartridge body also causes the insertion of the sample dispensing mechanism into the cartridge body.

4. 4. The sample inspection system of claim 1, wherein a single action by a user causes the sample dispensing mechanism to rupture the at least one seal and dispense the sample fluid from the cartridge body into the at least one reaction chamber.

5. the single action by the user is a sustained threading action applied to the closure relative to the cartridge body; the screwing action causes actuation of the sample dispensing mechanism and seals the cartridge body, the closure including threads. A sample inspection system according to claim 4 which cites claim 2 or 3.

6. The sample inspection system of claim 1 , wherein the at least one penetrating tip comprises a ball-point tip, an arrowhead tip, or a frusto-conical tip.

7. 4. The sample inspection system of claim 1, wherein a distal portion of the at least one penetrating tip includes a flat surface, the flat surface being at an angle of less than about 20°, about 15°, about 10°, about 5°, or about 1° relative to a surface of the at least one seal.

8. The sample testing system of claim 1 , wherein the at least one penetrating tip is fluted.

9. 4. The sample inspection system of claim 1, wherein the at least one penetrating tip comprises one or more flow channels, the one or more flow channels being positioned (i) at a proximal end of the at least one penetrating tip, (ii) at a distal end of the at least one penetrating tip, or (iii) along the length of the at least one penetrating tip.

10. 10. The sample inspection system of claim 9, wherein at least a portion of the predetermined partial volume of the sample fluid flows through the one or more flow channels and through the at least one opening, and the fluid flow is at a higher flow rate compared to a sample inspection system in which the at least one piercing tip does not include one or more flow channels.

11. 4. The sample testing system of claim 1, wherein the at least one piercing tip that ruptures the at least one seal comprises the at least one piercing tip that penetrates the at least one seal and moves into at least a portion of the at least one reaction chamber.

12. 12. The sample inspection system of claim 11, wherein the at least one opening expands in size as the at least one piercing tip moves into at least a portion of the at least one reaction chamber.

13. the at least one piercing tip rupturing the at least one seal can generate one or more flaps; the one or more flaps comprising one or more portions of the at least one seal ruptured by the at least one piercing tip; The sample inspection system according to any one of claims 1 to 3.

14. The sample testing system of claim 13 , wherein the flap does not adhere to the at least one piercing tip and / or does not obstruct fluid flow through the opening.

15. 4. The sample testing system of claim 1, wherein the piercing tip comprises a geometry configured to reduce wicking of the sample fluid into the at least one piercing tip and / or the one or more flaps.

16. 4. The sample testing system of claim 1, wherein the large opening comprises puncturing at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 99% of the surface area of ​​the at least one seal.

17. 4. The sample testing system of claim 1, wherein at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 99% of the surface area of ​​the at least one seal is in contact with the at least one penetrating tip.

18. 4. The sample inspection system of claim 1, wherein at least about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 98%, or about 99% of the predetermined partial volume of the sample fluid flows into the at least one reaction chamber.

19. 4. The sample inspection system of claim 1, wherein the predetermined partial volume of the sample fluid comprises at least about 10 μL, about 15 μL, about 20 μL, about 25 μL, about 30 μL, about 35 μL, about 40 μL, about 45 μL, about 50 μL, about 60 μL, about 70 μL, about 80 μL, about 90 μL, about 100 μL, about 110 μL, about 120 μL, about 128 μL, about 130 μL, about 140 μL, about 150 μL, about 160 μL, about 170 μL, about 180 μL, about 190 μL, or about 200 μL of the sample fluid.

20. The sample inspection system of claim 1 , wherein the sample dispensing mechanism includes an overmolded layer disposed on a surface of at least a portion of the dispensing rod.

21. 21. The sample inspection system of claim 20, wherein the overmolded layer forms a cylindrical seal.

22. 21. The sample inspection system of claim 20, wherein the overmolded layer comprises a thermoplastic elastomer (TPE) having a different durometer hardness than at least a portion of the dispensing rod, the overmolded layer exhibiting a Shore D durometer hardness or a Shore A durometer hardness of approximately 20-30.

23. the at least one reaction chamber is two reaction chambers; the at least one penetrating tip is a two-penetrating tip, and optionally, the reaction chamber comprises a polymerase chain reaction (PCR) tube, and further optionally, the two reaction chambers comprise mixing beads; The sample inspection system according to any one of claims 1 to 3.

24. 24. The sample testing system of claim 23, wherein the two reaction chambers contain different reagents selected to perform different respective tests and / or to detect different respective target entities.

25. the cartridge body contains a sample preparation reagent; At least one of the reaction chambers contains one or more reagents for a reverse transcription reaction and / or an amplification reaction; The sample inspection system according to any one of claims 1 to 3.

26. 5. The sample testing system of claim 4, wherein the cartridge body includes one or more alignment features configured to align with and engage one or more mating slots of a testing device, the one or more alignment features preventing rotation of the cartridge body when the cartridge body is in position on the testing device, and the one or more alignment features enabling a user to perform the single action with one hand.

Images