Diagnostic test assemblies, systems and methods

Abstract

A diagnostic test assembly for use in conducting biological or environmental tests. The diagnostic test assembly comprises a cartridge comprising: a cartridge body configured to house sample preparation fluid and configured to receive a biological or environmental sample through an opening, a reaction chamber comprising one or more amplification tubes, and one or more membranes configured to prevent fluid communication between the cartridge body and the one or more amplification tubes of the reaction chamber. The diagnostic test assembly comprises a dispense component configured to cooperate with the cartridge, wherein the dispense component comprises a securing portion configured to secure the dispense component to the cartridge and a dispensing portion configured to extend within the cartridge body of the cartridge when the dispense component is secured to the cartridge, and in response to actuation, the dispensing portion is configured to move relative to the securing portion.

Description

[0001] DIAGNOSTIC TEST ASSEMBLIES, SYSTEMSAND METHODS

[0002] Technical field

[0003] [1] Described embodiments relate to diagnostic test assemblies for use in conducting biological or environmental tests, instruments configured to cooperate with the diagnostic test assemblies to perform biological or environmental tests, and methods for conducting biological or environmental tests using the diagnostic test assemblies and instruments. Some embodiments relate to diagnostic test assemblies, instruments, systems, and methods for performing diagnostic tests and / or analysing biological samples to aid in environmental, agricultural, scientific, veterinary, and / or medical diagnoses based on detection of the presence or absence of one or more specific analytes in a sample and / or determining their quantities in the sample. The analyte may be detected using methods of molecular DNA amplification and detection of specific genetic markers, for example.

[0004] Background

[0005] [2] The amplification of nucleic acids is important in many fields, including medical, biomedical, environmental, veterinary and food safety testing. In general, nucleic acids are amplified by one of two methods: polymerase chain reaction (PCR) or isothermal amplification.

[0006] [3] Prior art diagnostic test system and apparatuses, in particular nucleic acid amplification and detection instruments are typically large, complex, and costly, and require sample preparation steps that must be conducted independently of the instrument. These preparation steps typically require a trained technical operator, and this operator and the test preparation environment can be exposed to hazardous samples such as body fluids and infectious agents, and the process is at risk from incorrect manual operations, including spills and incorrect reagent additions.

[0007] [4] The resulting test sample must then be accurately subsampled and transferred by a manual transfer step, typically a skilled pipetting operation. This approach requires a trained technical operator and a number of separate tubes and transfer devices, all of which will be contaminated by the sample and must be correctly handled and disposed of individually. [5] In these prior art approaches, the test sample is not sealed from the environment during the process of sample preparation and transfer into test tubes in the test instrument. This exposure to the sample can present infection agent risks to users and others, and can also contaminate the test instrument and test area, resulting in incorrect diagnostic results in subsequent tests.

[0008] [6] The diagnostic test system described in Australian Patent No. 2018340855 or in US Patent No. 11,709,175 addresses one or more of the above difficulties, the entirety of each are incorporated herein by reference.

[0009] [7] The diagnostic test system described in those patents comprises a diagnostic test apparatus (referred to as ‘instrument’) and a diagnostic test assembly. The diagnostic test assembly takes the form of a disposable diagnostic test cartridge that is produced prior to a diagnostic test, and already incorporates the precursor chemical components (i.e. reagents) to run a specific set of one or more diagnostic tests. The disposable diagnostic test cartridge is sealed with a shipping cap, and diagnostic the test assembly further comprises a dispense cap with an associated dispensing mechanism. The diagnostic test assembly is configured so that it can be safely handled without contamination from the environment or causing contamination of the user or the environment with the test materials or causing interference with these chemical components or otherwise affecting the subsequent operations of the cartridge, which require interactions with the diagnostic test instrument.

[0010] [8] In a typical test sequence, the diagnostic test cartridge body is preloaded with a volume of sample preparation fluid such as buffer and lysis liquid reagent and the separately sealed reaction tubes are pre-loaded with reagents to run a test such as isothermal DNA. This cartridge assembly is inserted into the instrument, and the instrument detects the presence of the diagnostic test cartridge and begins warming the sample preparation fluid. When the sample preparation fluid has reached a suitable temperature for sample addition, the instrument then prompts the user to remove the shipping cap and add the biological or environmental sample to be analysed.

[0011] [9] In the case of biological samples, the step of adding the sample to the sample preparation fluid within a sample preparation reservoir of the diagnostic test cartridge initiates a specific biological and chemical process that may include, PH buffering, specific salt concentrations, deactivation of inhibitors, sample dilution and cell lysis to prepare the sample material, including its included RNA or DNA nucleic acid, for testing. Typically, this sample preparation step requires a time duration at a controlled fluid temperature with the sample material present and can take many minutes of wait time.

[0012]

[0010] Subsequently, when adequate sample preparation time has elapsed, the user is prompted by the instrument software, to install the dispensing cap to the cartridge sample preparation reservoir, and the action of operating the closure not only seals the sample and sample preparation fluid within the cartridge, but also actuates a dispensing mechanism within the sample preparation reservoir to deliver a sub-sample of predetermined volume into one or more diagnostic test reservoirs within the cartridge.

[0013]

[0011] The instrument then controls the temperature of the one or more diagnostic test reservoirs and the sample fluid, and reagents contained within them. A cycle series or a time series of optical measurements of the contents of the test tubes is acquired by the instrument. The instrument processes these measurements to determine a test result which is provided as an output to a user.

[0014]

[0012] The sealed cartridge protects the reagents in transport and storage prior to running a test and supports the test process while the diagnostic test is underway. The test reagents, amplification genetic products, and contaminants are retained within the cartridge at all times, including at the completion of the test. The sealed cartridge can be removed for disposal at the completion of a test, and the instrument is protected from fluids and contamination at all times. Further, after the biological sample is added to and then sealed within the cartridge, the user is protected from the biological or chemical hazards of the sample during the subsequent test process and after the cartridge is removed for disposal.

[0015]

[0013] It is desired to address or ameliorate one or more of the disadvantages of the prior art, or at least to provide a useful alternative thereto.

[0016]

[0014] Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present disclosure as it existed before the priority date of each of the appended claims.

[0015] Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers, or steps, but not the exclusion of any other element, integer or step, or group of elements, integers, or steps.

[0017] Summary

[0018]

[0016] Some embodiments relate to a diagnostic test assembly for use in conducting biological or environmental tests, the diagnostic test assembly comprising: a cartridge comprising: a cartridge body configured to house sample preparation fluid and configured to receive a biological or environmental sample through an opening; a reaction chamber comprising one or more amplification tubes; and one or more membranes configured to prevent fluid communication between the cartridge body and the one or more amplification tubes of the reaction chamber; and a dispense component configured to cooperate with the cartridge, wherein the dispense component comprises: a securing portion configured to secure the dispense component to the cartridge; and a dispensing portion configured to extend within the cartridge body of the cartridge when the dispense component is secured to the cartridge, and in response to actuation, the dispensing portion is configured to move relative to the securing portion; wherein, when the dispense component is secured to the cartridge and the dispensing portion is selectively actuated to a first extent, the dispensing portion moves a first distance within the cartridge body, away from the securing portion toward the reaction chamber, and disrupts at least one of the one or more membranes, bringing the cartridge body into fluid communication with at least one of the one or more amplification tubes of the reaction chamber, and allowing fluid within the cartridge body to be dispensed from the cartridge body into the at least one of the one or more amplification tubes.

[0019]

[0017] The dispensing portion may be substantially elongate and is configured to move longitudinally relative to the securing portion.

[0020]

[0018] In some embodiments, securing the securing portion to the cartridge body does not cause actuation of the dispensing portion.

[0021]

[0019] In some embodiments, when the dispense component is fitted to the cartridge and the dispensing portion is selectively actuated to a second extent, the dispensing portion moves a second distance within the cartridge body away from the securing portion and without disrupting the one or more membranes to agitate fluid within the cartridge body, wherein the second distance is less than the first distance. Selective actuation of the dispensing portion to the second extent followed by return of the dispensing portion toward the securing portion may cause the dispensing portion to perform a reciprocal motion to thereby mix fluid within the cartridge body.

[0022]

[0020] In some embodiments, the dispensing portion comprises one or more piercing tips to disrupt the at least one of the one or more membranes, and wherein in response to actuation of the dispensing portion to the first extent, the one or more piercing tips of the dispensing portion protrude from the cartridge body and disrupt the at least one of the one or more membranes.

[0023]

[0021] The dispensing portion may comprise a plunger head connected to a plunger body, and wherein the one or more piercing tips protrude from the plunger body and wherein the plunger head is configured to be actuated to cause the dispensing portion to move relative to the securing portion. The plunger body may comprise one or more leg members, each leg member corresponding to a respective one of the one or more amplification tubes, and each leg member comprising a respective one of the one or more piercing tips, and wherein each of the one or more piercing tips is equipped with a tip seal to facilitate movement of the one or more piercing tips relative to internal walls of the amplification tubes when the dispense portion moves relative to the securing portion. The plunger head may define a first aperture in a side wall of the plunger head, the first aperture being arranged to receive an actuating member, which when it exerts a force, causes the dispensing portion to move relative to the securing portion.

[0024]

[0022] In some embodiments, the securing portion comprises a cap defining a chamber arranged to receive an upper section of the dispensing portion, wherein the cap defines a second aperture to provide access to the chamber, and wherein actuation of the dispensing portion can be performed through the second aperture. The first and second apertures may cooperate with one another such that both the first and second apertures are arranged to receive the actuating member therethrough to allow for actuation of the dispensing portion.

[0025]

[0023] In some embodiments, an end of the securing portion defines an opening configured to receive an upper portion of the plunger body such that the upper portion of the plunger head extends through the securing portion and is actuatable to cause the dispensing portion to move relative to the securing portion when the securing portion is attached to the dispensing portion.

[0026]

[0024] In some embodiments, the dispensing component is pneumatically actuated. For example, the dispensing portion may comprise a chamber connected to a plunger body, wherein an end of the securing portion defines an opening in fluid communication with the chamber of the dispensing portion, and where the dispensing portion is configured to be pneumatically actuated by the application of air through the aperture to pressurise the chamber.

[0027]

[0025] In some embodiments, the dispensing component is spring actuated. For example, the securing portion may comprise base portion housing a sliding bracket defining an elongate opening, wherein the elongate opening is relatively broad at one end and relatively narrow at another end, and wherein the dispensing portion comprises a rod connected to a spring and a plunger body, where the spring is biased towards the plunger body, and wherein an end of the rod is configured to be received by the elongate opening, and wherein when the sliding bracket is configured to transition, by way of a sliding motion, between an unactuated state, wherein the end of the rod is retained within the relatively narrow end of the elongate opening, and an actuated state, wherein the end of the rod is released through the relatively broad end of the of elongate opening, and is drawn away from the securing portion by action of the spring.

[0028]

[0026] The securing portion may comprise a screw thread configured to engage with a complimentary screw thread disposed on the cartridge body to thereby secure the dispense component to the cartridge.

[0029]

[0027] The securing portion may comprise one or more clips and / or protrusions configured to engage with complimentary clips and / or protrusions disposed on the cartridge body to thereby secure the dispense component to the cartridge.

[0030]

[0028] In some embodiments, the reaction chamber comprises one or more nozzles to improve placement of dispensed fluid in the one or more amplification tubes.

[0031]

[0029] The dispensing portion may be configured to be selectively actuated by a cooperating arm of an instrument configured to conduct the biological or environmental test using the diagnostic test assembly. The dispensing portion may be configured to be selectively actuated by an actuating member.

[0032]

[0030] The diagnostic test assembly may comprise a removable closure, such as a cap or foil closure, configured to close the opening of the cartridge body for storage and / or shipping of the cartridge.

[0031] In some embodiments, the cartridge body and the reaction chamber are welded together to form the cartridge. In some embodiments, the cartridge body and the reaction chamber each comprise complementary components to allow the cartridge body and the reaction chamber to clip together to form the cartridge. In some embodiments, the cartridge body and the reaction chamber each comprise complementary thread to allow the cartridge body and the reaction chamber to screw together to form the cartridge.

[0033]

[0032] At least one of the one or more amplification tubes may house preloaded dry or lyophilised reagents which are configured to dissolve when fluid is dispensed into the one or more amplification tubes. At least one of the one or more of the amplification tubes may house at least a preloaded magnetic mixing bead, which can be agitated by movement of one or more permanent magnets near the reaction chamber to mix content of the amplification tubes. The cartridge body may comprise the sample preparation fluid.

[0034]

[0033] The cartridge body and / or the reaction chamber and / or the one or more amplification tubes may be partially or entirely composed of a transparent material such that the contents thereof, and reactions that take place therein, can be observed visually or optically sensed by an instrument.

[0035]

[0034] The one or more membranes comprises: (i) a foil seal which covers all the one or more amplification tubes of the reaction chamber; or (ii) one or more foil seals, each of the one of more foil seals configured to cover a respective one of the one or more amplification tubes of the reaction chamber.

[0036]

[0035] Some embodiments relate to an instrument configured to perform a biological or environmental test using the diagnostic test assembly of any one of the described embodiments, the instrument comprising: a controller including one or more processors and memory, wherein memory comprises computer executable instructions for performing the biological or environmental test; a cartridge holder configured to receive the cartridge of the diagnostic test assembly; and an arm configured to selectively engage with the dispense component of the diagnostic test assembly to cause the dispensing portion of the dispense component to perform a mixing or dispensing function under the control of the controller.

[0037]

[0036] Some embodiments relate to a method of performing a biological or environmental test using the diagnostic test assembly of any one of the described embodiments. The method may further comprise providing a biological or environmental sample to the cartridge body, wherein the cartridge body comprises a sample preparation fluid; securing the dispense component to the cartridge; controlling, by the instrument, a temperature of the mixture in the cartridge body for lysis; causing an arm of an instrument configured to perform a biological or environmental test to actuate the dispensing portion to a first extent to cause the dispensing portion to move the first distance within the cartridge body, away from the securing portion toward the reaction chamber, and to disrupt the at least one of the one or more membranes, bringing the cartridge body into fluid communication with at least one of the one or more amplification tubes of the reaction chamber, and allowing fluid within the cartridge body to be dispensed from the cartridge body into the at least one of the one or more amplification tubes.

[0038]

[0037] In some embodiments, when using the diagnostic test assembly of any one of the described embodiments, controlling by the instrument, a temperature of the mixture in the cartridge body for lysis, further comprises: causing the arm of the instrument to actuate the dispense component to a second extent to cause the dispensing portion to move the second distance within the cartridge body away from the securing portion and without disrupting the one or more membranes to agitate fluid within the cartridge body.

[0039] Brief description of the drawings

[0040]

[0038] Embodiments of the present disclosure will now be described in further detail, by way of non-limiting example, with reference to the accompanying drawings, in which:

[0041]

[0039] FIG. 1 is an isometric view of a diagnostic test assembly comprising a closure, cartridge and dispense component, according to some embodiments;

[0042]

[0040] FIG. 2A is an isometric view of the dispense component of the diagnostic test assembly of FIG. 1 and FIG.2B is an expanded view of the dispense component of FIG. 1;

[0043]

[0041] FIG. 3A is an alternative expanded isometric view of the dispense component of FIGS. 2A and 2B, FIG. 3B is a cross-sectional view of the cartridge of FIG. 1, and FIG. 3C is a cross- sectional view of a portion of the cartridge of FIG. 1 with the dispense component secured thereto;

[0044]

[0042] FIG. 4 is an expanded isometric view of the cartridge of the diagnostic test assembly of FIG. 1;

[0043] FIG. 5A is an isometric cross-sectional view of a portion of the cartridge of Fig. 4, according to some embodiments and FIG. 5B is an isometric view of a reaction chamber of the cartridge of FIG. 5 A;

[0045]

[0044] FIG. 6 is an isometric view of a cartridge, having a screw thread and a corresponding closure, according to some embodiments;

[0046]

[0045] FIG. 7 is a nonlimiting example workflow depicting a diagnostic test assembly in use in a diagnostic test system, according to some embodiments;

[0047]

[0046] FIG. 8A is a cross sectional view of a cartridge fitted with a seal and an isometric view of a dispense component, according to some embodiments, FIG. 8B is a cross sectional view of the cartridge of FIG. 8A fitted with the dispense component of FIG. 8 A in an undispensed condition, and FIG. 8C is an isometric view of the cartridge of FIG. 8 A fitted with the dispense component of FIG. 8A in a dispensed state;

[0048]

[0047] FIG. 9 is a cross sectional view of a cartridge fitted with a dispense component in a dispensed condition, according to some embodiments, wherein a dispensing error has occurred;

[0049]

[0048] FIG. 10A is a cross sectional view of a cartridge comprising nozzles for targeted dispensing, fitted with a dispense component and in an undispensed state, according to some embodiments, and FIG. 10B is a cross sectional view of the cartridge and dispense component of FIG. 10(A), wherein the dispense component is in a dispensed state;

[0050]

[0049] FIG. 11 is an expanded isometric view of a portion of a clip-style cartridge featuring nozzles for targeted dispensing, according to some embodiments;

[0051]

[0050] FIG. 12A is a cross sectional view of a cartridge fitted with a dispense component in an undispensed / mixing state and an instrument arm engaging with the dispense component, and FIG. 12B is a cross sectional view of the cartridge fitted with the dispense component of FIG. 12A in a dispensed state and the instrument arm engaging with the dispense component;

[0052]

[0051] FIG. 13 is a cross-sectional view of a cartridge fitted with a dispense component, according to some embodiments, and wherein a lysis chamber of the cartridge contains a pre- loaded magnetic bead;

[0052] FIG. 14A is an isometric view of a cartridge fitted with a dispense component, with lysis chamber and reaction chamber heater blocks and FIG. 14B is a cross sectional view of the cartridge fitted with the dispense component of FIG 14B.

[0053]

[0053] FIG. 15A is an isometric view of an instrument including a cartridge fitted with a dispense component, wherein the instrument’s lid is open, and FIG. 15B is an isometric view of the instrument of FIG. 15Awherein the instrument’s lid is closed;

[0054]

[0054] FIG. 16 is a block diagram of a control system of the instrument of FIGS. 15A and 15B, according to some embodiments;

[0055]

[0055] FIG. 17A is a cross sectional view of a portion of the instrument of FIGS. 15A and 15B, wherein the instrument includes a cartridge fitted with a dispense component, wherein the instrument’s lid is open, and FIG. 17B is a cross sectional view of the portion of the instrument of FIG. 17A wherein the instrument’s lid is closed;

[0056]

[0056] FIG. 18A is a cross sectional view of an instrument including a cartridge fitted with a dispense component, wherein the instrument arm is being driven by a cartridge actuator motor to actuate a mixing function, and FIG. 18B is a cross sectional view of the instrument of FIG. 18A wherein the instrument arm is being driven by a cartridge actuator motor to actuate a dispense function;

[0057]

[0057] FIG. 19 is an isometric view of a portion of the instrument, according to some embodiments;

[0058]

[0058] FIG. 20 is a flow diagram of a workflow of the instrument, according to some embodiments;

[0059]

[0059] FIG. 21A is an isometric view of a cartridge fitted with a “push to activate” screw-on dispense component in an undispensed condition, and FIG. 21B is an isometric view of the cartridge fitted with a “push to activate” screw-on dispense component of FIG. 21A in a dispensed condition, and FIG. 21C is an expanded isometric view of the cartridge fitted with a “push to activate” screw-on dispense component of FIG. 21 A;

[0060]

[0060] FIG. 22A is an isometric view of a diagnostic test assembly comprising a pneumatically actuated dispense component fitted on a cartridge, and FIG. 22B is an isometric view of the dispense component wherein the dispense component has its cap removed; and

[0061] FIG. 23A is an isometric view of a portion of diagnostic test assembly comprising a cartridge fitted with a dispense component involving spring tension, wherein the upper cap is in place, FIG. 23B is an isometric view of the portion of diagnostic test assembly of FIG. 23A with the upper cap removed and in the undispensed state, and FIG. 23C is an isometric view of a portion of the diagnostic test assembly of FIG. 23 A with the upper cap removed and in the dispensed state.

[0061] Detailed description

[0062]

[0062] Described embodiments relate to diagnostic test assemblies for use in conducting biological or environmental tests, instruments configured to cooperate with the diagnostic test assemblies to perform biological or environmental tests, and methods for conducting biological or environmental tests using the diagnostic test assemblies and instruments.

[0063]

[0063] Some embodiments relate to a diagnostic test assembly for use in conducting biological or environmental tests. The diagnostic test assembly comprises a cartridge and dispense component configured to cooperate with the cartridge. The cartridge comprises a cartridge body configured to house sample preparation fluid and configured to receive a biological or environmental sample through an opening. The cartridge further comprises a reaction chamber having amplification tube(s). The cartridge further comprises membrane(s) configured to prevent fluid communication between the cartridge body and the amplification tube(s). The dispense component comprises a securing portion configured to secure the dispense component to the cartridge, and a dispensing portion configured to extend within the cartridge body of the cartridge when the dispense component is secured to the cartridge, and in response to actuation, the dispensing portion is configured to move relative to the securing portion.

[0064]

[0064] If fluid or a mixture, such as sample preparation fluid and a sample, is provided within cartridge body, fitting or securing the dispense component to the cartridge causes the opening to be closed, sealed or substantially sealed by the dispense component. The mixture can then be contained securely within the cartridge, with mixing and / or dispensing steps taking place internally, and without the need for any external fluid transfer steps to be taken.

[0065]

[0065] Thereafter, the mixture or fluid housed within the cartridge body can be selectively dispensed by actuating the dispensing portion of the dispense component. For example, fluid may be dispensed at a time that best suits the test being conducted and / or the operator’s preference or needs. When the dispensing portion is selectively actuated (for example, to a full extent), the dispensing portion of the dispense component moves a first distance within the cartridge body, away from the securing portion and toward the reaction chamber, and disrupts (for example, penetrates or dislodges) the membrane(s) of the cartridge. Disrupting the membrane(s) brings the cartridge body into fluid communication with the amplification tube(s) of the reaction chamber, allowing fluid within the cartridge body to be dispensed from the cartridge body into the amplification tube(s). In some embodiments, the dispense component may be selectively actuated (for example, to a partial extent) to cause the dispensing portion to move up and down (for example, in a reciprocating motion) within the cartridge body to induce mixing of the sample and sample preparation fluid within the sample fluid section of the cartridge body, but actuation travel limited to prevent causing the dispensing portion to disrupt the membrane(s) of the cartridge.

[0066]

[0066] Unlike the prior art diagnostic test system discussed above, the diagnostic test assembly of the described embodiments separates the acts of sealing the opening of the cartridge body (for example, containing the sample preparation fluid and a sample) and dispensing the contained fluid. In other words, securing the securing portion of the dispense component to the cartridge body does not cause actuation by the dispensing portion. This means that the sample can be supplied to the sample preparation fluid within the cartridge body and the dispense component fitted to the cartridge without fluid being dispensed. As a result, the assembled diagnostic test assembly (i.e., the cartridge fitted with the dispense component) can be inserted into the instrument to be heated to the desired temperature, and the dispensing portion then actuated (for example, automatically by the instrument) to dispense the fluid at a later time without any further intervention from the operator. This has several advantages. For example, a wait time between a first user interaction with the test system (i.e. the user inserting the cartridge into the instrument), and a second user interaction (i.e. the user adding a sample and fitting the seal and dispense cap to the cartridge, which directly results in two actions occurring - the cartridge being sealed and fluid being dispensed) is eliminated. In a typical test sequence using a test system comprising the prior art diagnostic test assembly, a user is required to wait approximately 3-4 minutes between the above first and second interactions while the instrument heats the preparation fluid to a desired temperature so that fluid is not dispensed before it is at the desired temperature. In this way, the dispensing actuation may be accurately timed and controlled by the instrument, without prompting the user to undertake the action. This may result in improved accuracy and reliability for the test reactions.

[0067] Furthermore, and as will be appreciated, in a busy laboratory setting, it is inefficient for a laboratory technician to wait for any period of time. In addition, a 3-4-minute wait time has been found to be particularly problematic, as operators such as laboratory technicians often attempt to address another task while waiting and become distracted. Delaying the fitting of the dispense cap and thereby the dispensing of the sample and sample preparation liquid can cause the reagents to denature, rendering the diagnostic test ineffective. The operator may then be required to restart the test with a new cartridge / reagents, which is costly in terms of both operator time and test materials.

[0067]

[0068] In some embodiments, the cartridge comprises one or more nozzles configured to extend toward or into respective amplification tube of the reaction chamber to provides targeted dispensing. For example, the provision of nozzles may cater for a more accurate quantity of fluid to be dispensed, improving the reliability of the diagnostic test assembly. More specifically, the provision of nozzles may mitigate or eliminate a problem of droplets of the fluid to be dispensed sticking to inner side walls of the cartridge body or reaction chamber, which may comprise sloped portions for example, and being prevented from reaching the amplification tube(s) of the reaction chamber where the test is performed. Many applications of the referenced test system involve dispensing small quantities of liquid, for example approximately lOOpL, and therefore a dispense discrepancy of only a few droplets can significantly affect the reproducibility and reliability of the test.

[0068]

[0069] Referring now to FIG. 1 there is shown an isometric view of a diagnostic test assembly 10, according to some embodiments. The diagnostic test assembly 10 comprises a cartridge 100 which is shown here fitted with a closure 110, such as a shipping cap or shipping closure, which may be a foil seal, for example. The cartridge 100 may be preloaded with a specific volume of sample preparation fluid, and suitable amplification and marker reagents provided within the reaction tubes for performing a predetermined set of one or more diagnostic tests.

[0069]

[0070] The cartridge 100 includes a cartridge body 120, which is configured to house the sample preparation fluid. The cartridge 100 comprises a reaction chamber 130, which is configured to house typically dried or lyophilised amplification and marker or fluorescence reagents. The walls of the reaction chamber 130 will typically be transparent to allow real time optical detection sampling measurements of the progress of the DNA using absorption, colour turbidity, visual and / or florescence detection modes.

[0071] The diagnostic test assembly 10 further comprises a dispense component 150 configured to cooperate with the cartridge 100.

[0070]

[0072] The cartridge body 120 and the reaction chamber 130 may be separate compartments of the assembly. In some embodiments, a membrane (not shown) is positioned between the cartridge body 120 and the reaction chamber 130 to prevent or inhibit fluid communication therebetween. The dispense component 150 is configured to cooperate with the cartridge 100 such that it can cause disruption to the membrane and thereby bring the cartridge body 120 and the reaction chamber 130 into fluid communication with each other.

[0071]

[0073] Typically, in use, a user places the cartridge 100 in an associated diagnostic test instrument or a standalone cartridge holder (not shown) and removes the closure 110. The user then adds a sample to the sample preparation fluid within the cartridge body 120 of the cartridge 110 and stirs the sample and sample preparation fluid. Users may choose to prepare the sample within a standalone cartridge holder instead of within the instrument to avoid the risk of any liquid being spilled on the instrument. Next, the user fits or attaches the dispense component 150 to the cartridge 100. For example, this may involve the user pushing a portion of the dispense component 150 into the cartridge body 120 of the cartridge 100 until secure; a click may indicate that the dispense component 150 is securely clipped in place.

[0072]

[0074] If the diagnostic test assembly 10 (including the cartridge 100 and dispense component 150) is not already in the instrument, the cartridge 100 (containing the sample and the sample preparation liquid and with the dispense component 150 attached) is inserted into the instrument, and the instrument heats the cartridge 100 to a desired temperature, where the cartridge body 120 and the reaction chamber 130 may be controlled at different temperatures to suit the specific reactions to be maintained in each volume. The dispense component 150 is then actuated automatically by the instrument to dispense a predetermined volume of the sample preparation fluid and sample mixture from the cartridge body 120 of the cartridge 100 into the reaction chamber 130. For example, actuation of the dispense component 150 can cause disruption to the membrane separating the cartridge body 120 and the reaction chamber 130 such that the sample preparation fluid and sample mixture flows from the cartridge body 120 into the reaction chamber 130. The reagents housed in the reaction chamber 130 then dissolve or are re-suspended into the fluid mixture, and further stages of the diagnostic test are then performed by the instrument. In some embodiments, the reagents in the reaction chamber may be stored as liquid and then simply mixed with the incoming dispensed sample fluid from the cartridge body 100.

[0073]

[0075] Notably, unlike the prior art, fitting the dispense component 150 to the cartridge 100 secures the dispense component 150 to the cartridge 100, acting as a closure for an opening of the cartridge body, and which may seal the cartridge, but it does not cause dispensing of the mixture from the cartridge body 120 of the cartridge 100 into the reaction chamber 130. A further actuating action is required to cause the dispense component 150 to dispense the mixture from the cartridge body 120. In some embodiments, the dispense component 150 is configured to interact with a portion of an instrument, such as an arm of the instrument, so that the instrument can selectively cause the dispensing to occur. Advantageously, once the sample has been added to the cartridge body 120 and the dispense component 150 is fitted to the cartridge 100, all further steps of a diagnostic test can be performed by the instrument, and there is no need for a user to wait to perform further steps.

[0074]

[0076] FIG. 2A is an isometric view of the dispense component 150 and FIG. 2B is an expanded view of the dispense component 150, according to some embodiments. Other embodiments of suitable dispense components are discussed below. FIG. 3 is an additional expanded isometric view of the dispense component 150, in which further features may be more readily apparent. The dispense component 150 comprises a securing portion 205, and a dispensing portion 207.

[0075]

[0077] The securing portion 205 is configured to secure the dispense component 150 to the cartridge 100. As illustrated, in some embodiments, the securing portion 205 comprises a cap 210. For example, the cap 210 may be substantially cylindrical. The cap 210 defines an open chamber 311 arranged to receive a part of the dispensing portion 207. Referring briefly to FIGS. 7E and 7F, the cap defines an aperture 380 within a wall of the chamber 311. The aperture 380 may extend longitudinally along a length or height of the cap 210. As discussed in further detail below, the open chamber 311 of the cap 210 may be configured to receive a part or upper section of the dispensing portion 207, and in response to actuation of the upper section of the dispensing portion 207 through the aperture 380 (for example, downward pressure exerted by an arm of an instrument), the dispensing portion 207 may be caused to move longitudinally, relative to and away from, the securing portion 205. By requiring actuation of the dispensing portion 207 to be performed through aperture 380, accidental or inadvertent dispensing of the mixture, for example, by user interaction, may be mitigated or circumvented.

[0078] The cap 210 may further comprise clips 220 which are configured to cooperate or interact with corresponding features of the cartridge body 120 when the dispense component 150 is fitted to the cartridge body 120. For example, when the dispense component 150 is inserted and pressed into place onto the cartridge 100, the clips 220 lock into place and secure and seal the dispense component 150 to the cartridge 100. Each clip 220 may extend from an end surface 203 of the cap 210. The clips 220 may be spaced apart from one another and may be disposed about a circumference of the cap 220. The closure portion 205 or cap 210 may comprise a cap detect tab 202. In some embodiments, an instrument sensor is configured to interact with the cap detect tab 202 to detect when the dispense component 150 and / or closure portion 205 is fully secured to the cartridge 100.

[0076]

[0079] The securing portion 205 may further comprise a cap seal 230 to improve the sealing function with the cartridge body 120.

[0077]

[0080] In some embodiments, as illustrated, the dispensing portion 207 of the dispense component 150 comprises a first dispensing part, such as a plunger head 240 and second dispensing part, such as a plunger body 250.

[0078]

[0081] When the dispense component 150 is secured to the cartridge 100, the securing portion 205 and the dispensing portion 207 may cooperate to close, and in some examples, seal or substantially seal, the opening of the cartridge body 120 of the cartridge 100. In some embodiments, the cap 210, plunger head 240 and seal 230 cooperate to provide the closing and / or sealing of the cartridge 100.

[0079]

[0082] The first dispensing part or plunger head 240 may be elongated, for example, to allow it to protrude up into the cap 210. As best shown in Fig. 3 A, the plunger head 240 defines an aperture 320 at or toward a first end. The aperture 320 may be disposed in a side wall of the first dispensing part 240. The aperture 320 is shaped to accommodate or receive a portion of an actuating arm of an instrument or a similar elongate object. By requiring actuation of the dispensing portion 207 to be performed through aperture 320 (alone, or in addition to aperture 380), accidentally or inadvertent dispensing of the mixture, for example, by user interaction, may be mitigate or circumvented. In other embodiments, the plunger head 240 does not include aperture 320 and the actuating arm, object or user’s finger acts or imparts pressure on the first end of the plunger head 240. Downward pressure on the plunger head 240 is effective to cause the plunger head 240 to move relative to the cap 210 to perform the dispensing action. Alternatively, a tool or a user’s finger, may be inserted into a suitably sized aperture 320 through the aperture 380 in the cap 210 to actuate the cartridge dispense if the cartridge is configured to be manually operated without instrument actuation.

[0080]

[0083] The first dispensing part or plunger head 240 is configured to connect or attach to the second dispensing part or plunger body 250. For example, and as illustrated, the plunger body 250 may comprise a clasp 360 disposed at or toward a first end of the plunger body 250 which is configured to engage with a corresponding feature 340 disposed at or toward a second end (opposite to the first end) of the plunger head 240 to join the plunger head 240 to the plunger body 250. The plunger body 250 may also comprise a protruding portion 235 configured to be received within a chamber defined by the plunger body 250, which may assist with improving the strength or robustness of the connection between the two parts. The attaching or connecting of the first and second dispensing parts is typically performed at the time of manufacture. In other embodiments, the plunger head 240 and body 250 are integrally formed.

[0081]

[0084] The plunger body 250 is configured to extend within the cartridge body 120. The plunger body 250 may be substantially planar. In some embodiments, the plunger body 250 may comprise tubular or rectangular section forms. The plunger body 250 may include strengthening ribs to assist injection moulding. The plunger body 250 may comprise ridges 280 running along a length of a first and / or second surface of the of the plunger body 250. The ridges 280 may improve the strength of the plunger body 250 and / provide extra rigidity so that the plunger body 250 does not deform as it moves into the cartridge 100.

[0082]

[0085] As mentioned above, the protruding portion 235 may extend from the first end of the plunger body 250. The protruding portion 235 couples with the plunger head 240 to form a full plunger assembly with a sliding seal on the plunger head 240 through the seal 230 into the cap 210. This assembly is made of two parts - the first dispensing portion or plunger head art 240 and second dispensing part or plunger body 250 - that clip together. This approach (two clip together parts) is used to assist with the complexity and cost of injection moulding and to assist with overall cartridge manufacture. However, the full assembly comprising the first dispensing portion 240 and the second dispensing portion 250 can be formed or moulded as a single part.

[0083]

[0086] One or more dispensing plunger portions 290 may extend from a second end of the plunger body 250, opposite to the first end. Each of the dispensing plunger portions 290 comprises a respective piercing tip 260 at its end. In some embodiments, one or more of the dispensing plunger portions 290 comprises a circumferential seal component such as an elastomer O-ring 270 (as shown in Fig. 3C).

[0084]

[0087] As the plunger body 250 is pressed down into cartridge body 120, the dispensing plunger portions 290 each form a seal with corresponding channel sections 481 disposed at or toward a base of the cartridge body 120. The channel sections 481 are shown in cross section in FIG 3B and FIG 4. The dispensing plunger portions 290 first seal a volume of fluid into the corresponding channel section 481 in the base of the cartridge body 120. This captured fluid is a known dispensing volume determined by the geometry of the channel section

[0085] 48 land the dispensing plunger portions 290. In this way, a defined sub sample volume of larger sample fluid volume in the cartridge 100 is isolated prior to dispensing into the corresponding amplification tubes 130. This captured volume of fluid is shown in FIG 3C. In this embodiment the sample fluid volume in the cartridge is 1.5mL and the captured subsample to dispensed into each of the lower reaction tube is lOOuL. As shown in FIG 3C, as the plunger portions 290 are further depressed by the actuation action applied to the cartridge 100, the respective piercing tips 260, pierce the membrane or foil 490 to provide a path for fluid captured within the channel section 481 to flow into the respective amplification tube 430 of the reaction chamber 130. The circumferential plunger seals 270 and the inner wall of the channel sections 481 interact similar to a syringe, that will force the fluid out of the channel sections 48 las the plunger portions 290 are fully depressed into the cartridge 100. In this way, the defined volume of fluid captured within the channel sections 481 is forced through the pieced passages membrane into the reaction tubes 130.

[0086]

[0088] The plunger body 250 of the exemplified embodiment of FIGS. 2A, 2B and 3 A has two legs 290 with two corresponding piercing tips 260, one for each of two amplification tubes of the reaction chamber 130 of the cartridge 100. As will be appreciated, in other embodiments, the plunger body 250 may have 1, 3, 4 or any other suitable number of legs 290 and associated piercing tips 260, configured to dispense liquid into a corresponding number of amplification tubes of a cartridge. Shown in FIG. 3C is a cross section of part of the cartridge 100 showing the piecing tips 260 and circumferential seals 270. The piecing tips 260 extend within the cavity of the channel sections 481 which holds a fixed volume of fluid in the cartridge body 120 prior to the piecing tips 260 being caused to pierce the membrane 490 and dispense the fluid into the reaction tube 130.

[0089] The seal 230 and the piercing tip seals 270 may be formed of rubber or a similar elastomeric material. The cap 210, plunger head 240, plunger body 250 and / or piercing tips 260 may be formed of rigid moulded plastic. The piecing tip seals 270 may be formed as cylindrical parts of the plunger portions 290 that form a close or tight fit with the inner wall of the channel sections 481 of the cartridge body 120. In some embodiments, the piecing tip seals 270 may be elastomeric plastic sections that are formed as a second over-moulding operation where these are injection moulded in a second operation over the first injection moulding that forms the plunger portions 290 and piecing tips 260.

[0087]

[0090] FIG. 4 is an expanded isometric view of the cartridge 100 of FIG. 1. The cartridge 100 comprises a cartridge body 120, a reaction chamber 130 and one or more membranes 490 located between the cartridge body 120 and the reaction chamber 130. The one or more membranes 490 are configured to prevent fluid communication between the cartridge body 120 and the one or more amplification tubes 430 of the reaction chamber 130 until it becomes disrupted, such as by being dislocated or pierced by the piercing tip(s) 260 of the plunger body 250.

[0088]

[0091] The reaction chamber 130 comprises one or more amplification tubes 430. In the example of FIG.4, the reaction chamber 130 houses two amplification tubes 430. As described above however, in other embodiments, the reaction chamber 130 may comprise 1, 3, 4, etc. amplification tubes 430. In this example, the reaction chamber 130 is substantially elongate, and comprises a substantially uniform diameter or width; in other words, the walls of the reaction chamber 130 do not taper significantly.

[0089]

[0092] The cartridge body 120 is configured to house sample preparation fluid and is configured to receive a biological or environmental sample through an opening. For example, the cartridge body 120 of the cartridge 100 may house a lysis chamber 480 into which sample preparation fluid may be the pre-loaded.

[0090]

[0093] The cartridge body 120 may comprise the one or more channel sections 481. Each channel section 481 may be configured to cooperate with a respective amplification tubes 430 of the reaction chamber 130. For example, each channel section 481 may feed into or be in fluid communication with a respective one of the two amplification tubes 430 once the membranes are disrupted. The channel sections 481 may be substantially elongate. The channel sections 481 may be substantially cylindrical, tubular and / or barrel shaped. In this example, the cartridge body 120 comprises two channel section 481, each cooperating with a respective one of the two amplification tubes 430.

[0091]

[0094] The cartridge body 120 is configured to receive the dispensing portion 207 of the dispense component 150. Accordingly, the cartridge body 120 is suitably shaped to accommodate the dispensing portion 207. In some embodiments, each channel section 481 is configured to receive a respective dispensing plunger portion 290 of the plunger body of the dispensing portion 207. For example, the dispensing plunger portion 290 extend within a respective channel section 481.

[0092]

[0095] In its non-actuated or non-dispensing state, the dispensing portion 207 is retained substantially within the cartridge body 120. When actuated, the dispensing portion 207 moves relative to the securing portion 205 and the piercing tips 260 of the plunger body 250 are caused to protrude from the cartridge body 120 to disrupt the membrane(s) positioned between the cartridge body 120 and the amplification tube(s) 430 of the reaction chamber 130.

[0093]

[0096] Typically, the cartridge body 120 and reaction chamber 130 are made of a transparent material so that the contents of the cartridge 100, as well as reactions that take place therein, can be observed visually or sensed by an instrument. In other embodiments, only a portion cartridge 100 and / or reaction chamber 130 is made of a transparent material. Atypical material used in the embodiments for these parts of the cartridge is polypropylene plastic as it can be injection moulded, can be formed in substantially transparent form for the reaction chambers and has good compatibility with the biological reagents and sample media used in the cartridge. Polypropylene plastic also performs well in the typical temperatures used in biological testing.

[0094]

[0097] In the exemplified embodiment, the cartridge body 120 has one or more ridges 412 which mate with corresponding one or more clips 220 disposed on the securing portion or cap 210 of the dispense component 150. For example, the cartridge body 120 comprises two ridges 412 disposed oppositely on an outer wall of the cartridge body 120. The ridges 412 may extend part way around a circumference of the cartridge body 120.

[0095]

[0098] The cartridge 100 may comprise an annular stabilising protrusion 470 to steady the cartridge 100 within the diagnostic test instrument and / or a freestanding holder. A locating feature 460 may be provided on the outside wall of the cartridge body 120 to ensure that the cartridge 100 is inserted into an instrument with the correct orientation.

[0099] In some embodiments, the cartridge body 120 and reaction chamber 130 are manufactured separately, and then joined after desired reagents are inserted into one or more of the amplification tubes 430.

[0096]

[0100] For example, a protrusion 420 disposed on an upper section of an outer wall of the reaction chamber 130 may be configured to cooperate with a mating recess or slot 410 disposed at a lower section of outer wall of the cartridge body 120 to allow the cartridge body 120 and reaction chamber 130 to clip together. This clip together functionality is particularly desirable in small-scale operations, where specialised methods of joining the two parts (120, 130) are cost prohibitive.

[0097]

[0101] The membrane(s) 490 may be attached to the base of the cartridge body 120 prior to the cartridge body 120 being loaded with sample preparation fluid or may be applied to openings of the amplification tube(s) 430 after the amplification tube(s) 430 are loaded with the relevant reagents. In some embodiments, the membrane(s) 490 is formed of foil. However, it will be appreciated that any other suitable material known to the skilled person that is capable of prohibiting fluid communication until being pierced or otherwise disrupted by the piercing tips 260 may be used. In some embodiments, multiple separate membranes 430 may be used to seal one or more of the amplification tubes 430 separately.

[0098]

[0102] In some embodiments, a barcode is adhered, printed or etched onto a surface of the cartridge 100 to identify one or more of: the test type to be performed, a batch or lot number, expiry date of the cartridge, lot-specific calibrations etc. In other embodiments, rather than a barcode, the identifying marker may instead comprise of a 2D quick response (QR) code or any other suitable unique identification code or schema known to the skilled person.

[0099]

[0103] FIG. 5 A is an isometric cross-sectional view of a portion of a cartridge 500 and FIG 5B is an isometric view the reaction chamber of a cartridge 500, according to some embodiments. In this example, the cartridge body 520 and reaction chamber 530 are designed to be welded together at a weld interface 510 to form a welded cartridge, instead of clipping together as per the embodiment of FIG. 4. This welded configuration may be more suitable for large-scale production of loaded cartridges 500, where more sophisticated machinery may be available. As the skilled person will appreciate however, there will be many other suitable ways in which the cartridge body 120, 520 and reaction chamber 130, 530 can be joined together. For example, the cartridge body 120, 520 and the reaction chamber 130, 530 may each comprise a complementary thread to allow the cartridge body and the reaction chamber to screw together to form the cartridge.

[0100]

[0104] FIG. 6 shows a cartridge 600 according to some embodiments. The cartridge 600 includes a screw thread 620. In this example, the screw thread 620 is disposed about an outer surface of a protruding portion of the cartridge body 630. The screw thread 620 is arranged to cooperate with a complimentary thread disposed on a closure 610 such as a shipping closure or shipping cap 610. In this example, the complimentary thread (not shown) is disposed about an inner surface of closure 610. As will be appreciated, any other suitable closure 610 can be used to seal the loaded cartridge 100, 500, 600 for storage and / or shipping prior to use. An associated alternative dispense component (not shown) can be configured to screw onto the cartridge 600, rather than clipping on.

[0101]

[0105] FIG. 7 is a nonlimiting example workflow 700 for a diagnostic test assembly 10 according to some embodiments, being used with a diagnostic test instrument in a diagnostic test system. The dashed box 701 indicates steps which are performed by a user / laboratory technician (A-D), while the remaining steps are performed automatically by a diagnostic test instrument (not shown). Typically, steps A-D will be performed with the cartridge resting inside the instrument, however in some applications, the cartridge 100 may be placed in a separate holder while the sample preparation steps are performed.

[0102]

[0106] The first step of the workflow (A) shows pre-loaded cartridge 100 with a shipping closure / foil seal 110 attached thereto. The user peels off the closure 110 (B) and uses a swab 710 to introduce a sample into the pre-loaded sample preparation fluid of the lysis chamber 480 within the cartridge body 120. The swab 710 is used to stir the sample and the sample preparation fluid within the cartridge volume.

[0103]

[0107] The dispense component 150 is then fitted to the cartridge 100, with the dispensing portion 207 being inserted into the cartridge body 120 and clicked in place within the cartridge 100 (D). The instrument (not shown) then heats the preparation fluid within the lysis chamber 480 of the cartridge body 120 to a desired temperature. During the heating stage, an arm of the instrument (not shown) makes small vertical movements within the aperture 320 of the plunger head 240, causing the plunger body 250 to move longitudinally relative to the securing portion 205 of the dispense component 150 to agitate or cause fluid mixing of the sample preparation fluid and sample mixture, with a view to keeping the mixture substantially homogenous or mitigating against sediment accumulation at the base of the lysis chamber 480. Movement of the instrument arm within the aperture 320 is represented by the arrow 720 (E).

[0104]

[0108] After the desired temperature is reached, the instrument arm actuates the dispense component 150 by making a larger downward movement within the aperture 320 of the plunger head 240 (as indicated by the arrow 730 (F)), causing the plunger body 250 to move a great longitudinal distance and causing the piercing tips 260 of the plunger body 250 to protrude from the cartridge body and pierce the membrane 490, thereby bringing the cartridge body 120 and reaction chamber 130 into fluid communication with one another. This allows movement of fluid from the lysis chamber 480 of the cartridge body 120 into the amplification tubes 430 of the reaction chamber 130. Pre-loaded reagents of the amplification tubes 430 then dissolve into the dispensed mixture.

[0105]

[0109] Dots 740 represent the remaining steps, which are completed by the instrument, and may include, for example: further heating as the instrument controls the temperature of the amplification tubes 430 and the sample fluid and reagents contained within them; obtaining optical measurements of the contents of the test tubes acquired by the instrument; and / or processing the measurements to determine a test result, which can be provided as an output to a user. In some embodiments, the amplification tubes 430 may also house pre-loaded magnetic mixing beads 810, which can be agitated by the instrument to cause mixing of the fluid within the amplification tubes 430 and help the lyophilised regent beads to dissolve into the fluid. The instrument agitates the mixing beads by moving a permanent magnet or otherwise creating a changing electromagnetic field close to the amplification tubes 430.

[0106] [HO] FIGS. 8A to 8C provide further detail regarding the dispensing action of the dispense component 150, within a cartridge 500. Referring to FIG. 8 A prior to its use, each of the amplification tubes 430 of the cartridge 500 contain a mixing bead 810 and a lyophilised pellet 820, and the lysis chamber 480 contains approximately 1.5mL of sample preparation liquid 850. The mixing bead may of a ferrous material or alloy or magnetically attracted material such that it can be influenced and moved by an external magnetic field to induce mixing of the fluid and components contained within the reaction tube.

[0107] [Hl] In use, the closure (e.g., a foil seal) 110 is removed, a sample is added to and mixed into the sample preparation liquid 850 in the lysis chamber 480, the dispensing portion 207 is inserted into the cartridge body and the dispense component 150 is secured to the cartridge 500. Clips 220 of the securing portion 205 (for example, cap 210) of the dispense component 150 engage with complimentary ridges 412 of the cartridge 500 to secure the dispense component 150 to the cartridge 500, as shown in FIG. 8B. The double ended arrow 830 shows a pre-dispensed liquid volume of just over approximately 1.5mL, i.e. the sample and sample preparation liquid mixture 860 within the lysis chamber 850. The liquid 860 has been displaced slightly by the plunger body 250 of the dispense component 150 however it remains housed within the lysis chamber 580, as the membrane 490 of the cartridge 500 is not yet disturbed.

[0108]

[0112] After heating has occurred, an arm of the instrument actuates the dispensing portion 207 by moving downwards within the aperture 320 of the plunger head 240 of the dispense component 150 and exerting a force on the plunger head 240. In response, the plunger body 250 moves down causing the piercing tips 260 of the plunger body 250 to protrude from the cartridge body 120 and disturb, for example, pierce, the membrane 490, allowing a predetermined volume (for example, approximately lOOpL) to move from the lysis chamber 580 into each of the amplification tubes 430, as shown in FIG. 8C. This approximately lOOpL of dispensed liquid is indicated by the double ended arrow 840.

[0109]

[0113] It has been found in practice that in some applications, due to the shape of the amplification tubes 430, the quantity of liquid dispensed from the lysis chamber 480 into the amplification tubes 430 can vary due to small droplets of liquid becoming stuck to the upper walls of the reaction chamber 130, 530. Typical uses of the cartridge 100, 500, 600 involve dispensing small volumes of liquid, and therefore any slight variation in the quantity of liquid dispensed into the amplification tubes 430 may be undesirable.

[0110]

[0114] FIG. 9 is a cross sectional view of a cartridge 500 fitted with a dispense component 150 in a dispensed state, where, due to the shape of the sloped portion 930 of the reaction chamber 530, the desired quantity of the sample and sample preparation liquid mixture 860 has not fully dispensed into the amplification tubes 430. A droplet of dispensed liquid 920 clings to the sloped portion 930 of the amplification tubes 430.

[0111]

[0115] FIGS. lOA and 10B are cross sectional views of a cartridge 1000 fitted with a dispense component 150 configured to provide targeted dispensing, according to some embodiments. FIG. 10A shows the cartridge 1000 and dispense component 150 in an undispensed state and FIG. 10B shows the cartridge 1000 and dispense component 150 in the dispensed state. As illustrated, in some embodiments, the reaction chamber 130 may comprise relatively straight walls. Straight walls as opposed to tapered walls, for example, may mitigate droplets attaching to or getting stuck to inner walls or surfaces of the reaction chamber 130. Here, at least one of the one or more amplification tube 430 of the reaction chamber of cartridge 1000 is equipped with a nozzle 1010 to direct dispensed fluid into a respective amplification tubes 430, helping to keep the dispensed fluid away from the upper inner walls of the amplification tubes 430. In some embodiments, the nozzles 1010 are configured to extend into the amplification tubes 430 to thereby improve placement of the dispensed fluid in the amplification tubes 430. The nozzles 1010 are therefore configured to receive the piercing tips 270 of the plunger body 250 when the dispense component is actuated (to a dispense state). In the dispensed state, as illustrated in FIG. 10B, the piercing tips 270 of the plunger body 250 have moved into the nozzles 1010. The nozzles 1010 may improve the reliability of the quantity of liquid dispensed into the amplification tubes 430. In other embodiments, nozzles 1010 may only be present in one or more selected amplification tubes 430, rather than in all the amplification tubes.

[0112]

[0116] FIG. 11 provides an expanded isometric view of a portion of the cartridge 1000 with nozzles 1010. This example, shows the cartridge 1000 as being essentially a clip-style cartridge 100 with a nozzle piece 1150 comprising the nozzles 1010 which is received by (for example, slides into) the amplification tubes 430 of the reaction chamber 130. The membrane 490 is applied either to an upper surface of the nozzle piece 1150 or to a lower surface or base of the cartridge body 120. As will be appreciated, the nozzle portion 1150 may be integrally or separately formed as shown in FIG. 11 but may also be used with a welded cartridge or any other similar cartridge, for example the cartridge 500, or any other of the described cartridges.

[0113]

[0117] Alternatively, or in addition to using nozzles 1010 to direct fluid flow, a hydrophobic material such as a suitable form of polypropylene plastic can be used to form the walls or inner surfaces of the amplification tubes 430. A coating or surface treatment may be applied to the inside walls of the reaction tubes to improve hydrophobic properties of these surfaces to cause any the droplets to not be strongly attached by surface tension and to flow down to the fluid volume at the base of the tube reaction tubes 430. The use of plasma treatment of the inside tube walls, or use of a plasma applied coating may be used to treat or coat the inner surface of the amplification tubes 430, such as part or all of the inner walls of the amplification tubes 430 to affect the hydrophilic surface tension of the inner surfaces of the reaction chamber 130, 530 to mitigate or prevent droplets forming on upper inner surfaces of the amplification tubes 430.

[0118] As described above regarding FIG. 7, in some applications, an arm of the instrument may be used to actuate the cartridge to induce stirring in the sample and sample preparation liquid mixture 860 during or after heating and prior to dispensing, to prevent sediment buildup at the base of the lysis chamber 480 and to produce an ideally homogenous dispensed liquid.

[0114]

[0119] FIG. 12Ais a cross-sectional view of an assembled diagnostic test assembly comprising a cartridge 1000 fitted with a dispense component 150 in a mixing but undispensed state and FIG. 12B is the cross-sectional view of an assembled diagnostic test assembly in a dispensed state. In both figures, an arm 1215 of an instrument (not shown) is inserted through the aperture 380 of the securing portion or cap 210 and into the aperture 320 of the first dispensing part or the plunger head 240 of the dispense component 150. The remainder of the instrument is not shown for simplicity.

[0115]

[0120] As represented by the arrow 1230 in FIG. 12A, the instrument arm 1215 is configured to make small vertical movements to agitate the sample and sample preparation mixture 860. Movement of the mixture 860 is represented by the mixing arrows 1220, which are indicative of fluid flow during the mixing. To perform the mixing function, the instrument is configured to cause the instrument arm 1215 to selectively actuate the dispensing portion 207 of the dispense component 150 to cause the dispensing portion 207 to move across a first distance or mixing range, for example, by performing a reciprocating motion, such as up and down. For example, the dispense component may be selectively actuated to a partial extent (such that the dispensing portion 207 moves only part way of its full range of motion), and not to a full extent. The partial extent may allow the dispensing portion 207 to travel a second distance within the cartridge body and away from the securing portion but be insufficient to disrupt the membrane(s). That is the actuation travel of the dispensing portion 207 may be limited to prevent disruption of the membrane(s) of the cartridge.

[0116]

[0121] When being selectively actuated (back and forward or up and down) to a partial extent, the dispense component 150 is operating in a mixing state. As shown in FIG. 12B, once the desired heating / mixing has occurred, the instrument then makes one larger vertical movement as indicated by the arrow 1240 to cause the dispense component 150 to actuate. To perform the dispensing function, the instrument is configured to cause the instrument arm 1215 to selectively actuate the dispensing portion 207 of the dispense component to cause the dispensing portion 207 to disrupt the membrane(s) bringing the lysis chamber of the cartridge body and the reaction chamber into fluid communication with one another and thereby causing the dispensing of the mixture into the reaction chamber. For example, the dispense component may be selectively actuated to a full extent to perform the dispensing function. The full extent may allow the dispensing portion 207 to travel a first distance within the cartridge body and away from the securing portion such that the dispensing portion 207 disrupts the membrane(s). When actuated to a full extent, the dispense component 150 is operating in a dispensing state.

[0117]

[0122] In some embodiments, other methods may be used (alternatively or in addition to) to mix the sample and sample preparation mixture 860. For example, referring to FIG. 13, a preloaded mixing bead 1316 is provided within the lysis chamber 480. This mixing bead 1316 may contain ferrous alloy or magnetically attracted material such than it can be acted on by a magnetic field provided by the instrument but external to cartridge body. The magnetically attracted mixing bead may be acted on by a magnetic field or permeant magnet within the instrument (not shown), where this field is moved or varied in strength to induce movement of the metallic bead and induce a stirring action within the cartridge fluid. In further embodiments still, any other method of mixing known to the skilled person may be utilised.

[0118]

[0123] FIG. 14A is an isometric view and FIG. 14 B a cross sectional view (B) of a cartridge 1000 fitted with a dispense component 150 and inserted into two heater blocks, for example, electrically heated aluminium blocks of the instrument. The heater blocks comprise a lysis chamber heater block 1410 and a reaction chamber heater block 1420. Again, the rest of the instrument is not shown for simplicity. The locating feature 460 of the cartridge 1000 slots into a corresponding recess 1460 of the lysis chamber heater block 1410, ensuring that the cartridge 1000 has been inserted with the correct orientation. The reaction chamber heater block 1420 has an apertures 1430 arranged to receive a corresponding amplification tube 430, and through which the instrument can detect and measure fluorescence using a camera. These heater blocks may be controlled at different temperatures using electric heating and temperature feedback with the instrument (not shown), and varied in temperature over time to provide required temperature conditions for diagnostic reactions within the cartridge body and the reaction tubes.

[0119]

[0124] FIGS. 15A and 15B show isometric views of a diagnostic test apparatus or instrument 1500 used to conduct diagnostic tests, according to some embodiments. The instrument 1500 comprises a lid 1520, for example, a sliding cover, arranged to transition between an open state, as illustrated in FIG. 15 A, and a closed state, as illustrated in FIG. 15B. The arrow 1510 indicates the direction in which the lid 1520 slides when moving from the open state to the

[0120] T1 closed state. In some embodiments, the lid 1520 is manually activated, allowing a user to manually manipulate the lid to cause it to move between the open and closed states, for example, using a sliding motion. In some embodiments, the lid 1520 is electrically actuated by an electric motor which drives a pinion gear against an internal rack gear in the lid 1520, the lid sliding within linear guides or a motor. In another implementation, a motor, pulley, and cord combination may be used to ‘wind in’ the lid. In some embodiments, the lid 1520 is held by spring force in the closed position when the instrument is not in use to protect a cartridge insertion port from environmental contamination. To commence a test, the user will slide the lid 1520 back against the spring force and the lid 1520 will latch in the open position using a mechanical or magnetic latch. In the open position, the cartridge insertion port area is open to allow insertion of the cartridge and the operation of fitting the dispense cap assembly. Once the dispense cap is fitted, the instrument can release the lid to move back to the closed position automatically to run the test, or can prompt the user to manually close the lid.

[0121]

[0125] As illustrated, in some embodiments, a front face of the instrument 1500 includes an on / off button 1530, a USB port 1540 and a light emitting diode (LED) to indicate whether the instrument has power and / or the current instrument status, for example, whether the instrument is ON, OFF, or on standby.

[0122]

[0126] The instrument 1500 further comprises a rigid enclosure or housing 1560 which may be constructed of, for example, moulded plastic or metallic sheet materials. A touch sensitive LCD display 1550 may be mounted within the housing 1560. The instrument 1500 is operated by selecting touch sensitive controls on a user interface (not shown) rendered on the instrument display 1550. However, in other embodiments, the user interface may be implemented as physical controls or buttons each having corresponding functions, or may be voice- controlled / operated.

[0123]

[0127] FIG. 16 is a block diagram of a control system of the instrument 1500. The instrument 1500 comprises a processing unit 1600, external data interfaces 1606, an optical measurement unit 1607 and a cartridge identification, heating, and actuation unit 1609.

[0124]

[0128] The processing unit 1600 comprises a microprocessor-based controller 1601 configured to execute diagnostic test processes that control the sequencing and operation of the instrument’s electrically operated functions. In the described embodiments, the microcontroller 1601 is configured by way of an embedded operating system 1602 and embedded software 1603 stored in non-volatile ‘data storage’ memory 1604, and a random-access memory (RAM) 1605 for temporary data storage, for example for storing raw data values during test result calculations. The non-volatile memory 1604 also stores test outcomes and instrument calibrations as data stored in one or more data files or in a database, and the information and data stored in the non-volatile memory is retained, even when no power is provided to the diagnostic test instrument 1500.

[0125]

[0129] The processing unit 1600 may be in communication with at least one external data processor via one or more data interfaces 1606 for example, a serial port, universal serial bus (USB) port, ethernet port or wireless access port (WAP) for connection to a Wi-Fi network. Additionally, the processing unit 1600 is in communication with the optical measurement unit 1607 and the cartridge identification, heating, and actuation unit 1609 via an internal control communications and I / O bus 1608.

[0126]

[0130] The optical measurement unit 1607 may comprise two optical measurement sensors: optical measurement sensor A 1627 and optical measurement sensor B 1628, which are used to acquire optical fluorescence signals from each of the amplification tubes 430 through the apertures 1430 in the side of the reaction chamber heater block 1420. The unit 1607 further comprises a motor 1626 which allows the optical measurement sensors 1627, 1628 to be accurately positioned with respect to the amplification tubes 430 to acquire measurements that represent the DNA or RNA detection probe emissions within the amplification tubes 430, as well as be moved to self-test reference locations and to park the sensors away from the location of the amplification tubes 430 when they are not required. The location of the measurement sensors 1627, 1628 is detected by a motor carriage position sensor 1625 and provided to the micro-controller 1601 as feedback to enable precise positioning.

[0127]

[0131] In some embodiments, the cartridge identification, heating, and actuation unit 1609 comprises a lid position sensor 1622, a ‘cartridge inserted’ sensor 1623, an internal barcode reader 1615, a dispense component fitted sensor 1624, a cartridge actuator / dispense motor 1610, and a lysis chamber heater block 1410 and reaction chamber heater block 1420 with associated respective heater elements 1620, 1621 and temperature sensors 1618, 1619.

[0128]

[0132] Opening the lid 1520 activates the lid position sensor 1622 such that the software application 1603 running on the instrument controller 1601 is aware that a test is starting and provides suitable prompts and feedback to the user. Opening and closing of the lid may be performed manually by a user or actuated by the controller 1601 either in response to a user input, or as an action in the workflow being performed by the instrument 1500.

[0129]

[0133] The cartridge 100, 500, 600, 1000 is then inserted into the instrument 1500 by the user. Once inserted, the cartridge is detected by the ‘cartridge inserted’ sensor 1523. In one implementation, the sensor comprises a light emitting diode (LED) forming a narrow beam across the cartridge port and associated photoelectric sensor that receives and detects the beam. The presence of an inserted cartridge breaks this beam and allows the instrument and its controller and software to detect the presence of the cartridge. Other suitable sensors include those that rely on detection of optical reflection from the surface of the cartridge, or the use of a switch that is activated by the cartridge insertion or the use of a camera and associated image analysis in software to confirm the presence or absence of the cartridge 100, 500, 600, 1000.

[0130]

[0134] The barcode reader 1615 then reads a barcode which is adhered, printed, or etched onto the surface of the cartridge 100, 500, 600, 1000, or alternatively where the workflow allows, on the shipping closure 110, 610. The diagnostic test system can be configured to process different test types, wherein these different tests can have different reagent contents, different intended target diagnostic test results and require different test processing conditions. The barcode identifies the cartridge type and test type to be completed. The barcode typically also provides the batch or lot number and the expiry date of the particular cartridge. This data can be used to apply lot-specific calibrations, and to exclude expired cartridges from being used in tests. This information is also included as metadata in the test result data generated by the controller during the diagnostic testing.

[0131]

[0135] The dispense component sensor then determines whether the dispense component 150 has been fitted. As described above, a user may prepare a sample - i.e. remove the shipping closure and add a sample to the sample preparation liquid in either in a freestanding cartridge or while the cartridge is resting within the instrument. Once the sample is added, the dispense component 150 is fitted, at which time the cartridge 100, 500, 600, 1000 is ready for the heating stage to commence. Hence, the instrument dispense component sensor 1624 provides feedback to the micro-controller that the test can commence. At this stage, a user may be promoted to close the lid 1520.

[0132]

[0136] As described with reference to FIG. 14, the instrument may comprise two heater blocks: the lysis chamber heater block 1410 and reaction chamber heater block 1420. The lysis chamber heater block 1410 and reaction chamber heater block 1420 may operate independently and are sufficiently insulated within the instrument such that they can operate at different temperatures. Each heater block 1410, 1420 may be heated using a respective heater element 1620, 1621, may be controlled separately by the instrument controller 1610, and may have an attached respective feedback temperature sensor 1618, 1619, which provides temperature readings to the controller 1610. The instrument controller 1610 heats and predicts the temperature imparted to the fluids within lysis chamber 480 and amplification tubes 430 of the reaction chamber 130, 530. Alternative implementations may use heated air chambers or any other method of directly or indirectly heating or cooling the fluids in the lysis chamber. Further, in other implementations, the temperature sensors may be of a non-contact type such as infrared temperature emission sensors, used to directly measure the temperature of the internal cartridge fluids, wherein the walls of the cartridge are constructed from materials with suitable transmission at the wavelengths needed to undertake this direct internal temperature measurement.

[0133]

[0137] In a typical application, the lysis chamber heater block 1410 operates from 40°C to 95°C, dependent on the type of sample to be processed and the cell types and cell lysis required. In a typical isothermal amplification application, the reaction chamber heater block 1420 operates at a fixed temperature such as 65°C. In a typical PCR application, the reaction chamber heater block 1420 heats and cools in steps between set temperatures over a typical range of 50°C to 95°C.

[0134]

[0138] During heating of the lysis chamber 480 by the lysis chamber heater block 1410, the dispense component 150 of the diagnostic test assembly is actuated by the arm of the instrument in a way to cause the dispensing portion 207 to adopt the mixing state, and move longitudinally (for example, up and down) within the cartridge body to mix the mixture in the lysis chamber 480. For example, in response to receiving instructions from the controller 1601, the cartridge actuator motor 1610, in combination with a spring, may drive the arm 1215 of the instrument 1500 to make small vertical movements (see FIG. 12, FIG. 18) to agitate the sample preparation fluid and sample mixture with a view to maintaining a homogenous mixture, and mitigating or preventing sediment accumulation at the base of the lysis chamber 480. Once this heating stage is complete, the dispense component 150 of the diagnostic test assembly is actuated by the arm of the instrument in a way to cause the dispensing portion 207 to adopt the dispensing state, and causing the dispensing portion 207 to move longitudinally and protrude from the cartridge body to disrupt the membrane(s) and dispense the mixture into the reaction chamber. For example, in response to receiving instructions from the controller 1601, the cartridge actuator motor 1610 in combination with a spring may drive the arm 1215 of the instrument 1500 to actuate the dispense component to cause the dispensing function. Following the dispense, the reaction chamber heater block 1420 begins to heat the amplification tubes 430.

[0135]

[0139] While the amplification is running, test readings are taken by the optical sensors 1627, 1628. These sensors are configured to have an optical path that incorporates the contents of the amplification test tubes through apertures 1430 in the reaction chamber heater block 1420 and corresponding optically transparent sections on the amplification tubes 430 / reaction chamber 130, 530. Where detection is required at amplification tubes 430 and at reference self-test regions, a scanning approach is used to move the sensors across these multiple detection points and provide measurements at each point of interest for the diagnostic test. These measurements can use optical absorption, fluorescence emission or bioluminescence emission to detect specific biological or genetic sequence markers in the amplification tubes 430. In other implementations, sensors with alternative measurement methods can operate in the same instrument 1500 and with the diagnostic test cartridges 100, 500, 600, 1000 described herein. These sensors can use magnetic, electrical, atomic, or physical properties of the test fluids to acquire measurements suitable to determine a test result.

[0136]

[0140] While a micro-controller based instrument 1500 is described, it will be apparent to those skilled in the art that some or all of the steps of the diagnostic test process performed by the instrument 1500 can alternatively be implemented in other forms, such as configuration data for a field-programmable gate array (FPGA), or entirely in hardware form, such as an application-specific integrated circuit (ASIC), for example.

[0137]

[0141] In some applications, the cartridge 100, 500, 600, 1000 and dispense component 150 can be used manually without an instrument 1500, where the cartridge is held in one hand, and the shipping closure 110, 610 is removed with the other hand. The sample is then added and the dispense component 150 fitted. In this case, where the amplification tubes 430 are visually transparent, the dispensing of fluid into the amplification tubes 430 can be visually observed, and a colour or turbidity change observed over time to produce a diagnostic test readout or display. In some embodiments, once the dispense component 150 is secured to the cartridge 100, 500, 600, 1000 after the sample has been added, the cartridge is sealed and the dispense component 150 actuated to optionally mix, and internally dispense a measured volume of diluted, prepared sample fluid into the test tube without the use of external fluid transfer steps. Further, a cartridge 1000 fitted with nozzles 1010 provides targeted dispensing, wherein a more accurate quantity of liquid is dispensed, when compared with the prior art described herein.

[0138]

[0142] FIGS. 17A and 17B are cross sectional views of a portion of an instrument 1500 including a diagnostic test assembly, comprising a cartridge 1000 fitted with a dispense component 150, where the instrument’s lid 1520 is open and closed, respectively. In this example, closing the lid 1520 causes a rod 1715 on which the arm 1215 of the instrument 1500 pivots, to slide within a guiding rail 1714 in the direction of the arrow 1640 so that the arm 1215 moves inside the aperture 320 of the cap 210 of the dispense component 150. In some embodiments, movement of the arm 1215 within the dispense component 150 is controlled by the cartridge actuator / dispense motor 1610 in combination with a spring (not shown).

[0139]

[0143] FIGS. 18A and 18B are cross sectional views of a portion of the instrument 1500 including a diagnostic test assembly, comprising a cartridge 1000 fitted with a dispense component 150, wherein the instrument arm 1215 is being driven by an actuator, for example, a liner actuator such as the cartridge actuator motor 1610 and a spring (not shown), to mix a mixture in the lysis chamber of the cartridge body, and to actuate a dispense from the lysis chamber to the reaction chamber.

[0140]

[0144] In some embodiments, to perform the mixing, the controller 1601 causes the cartridge actuator motor 1610 to turn a leadscrew 1809 in a direction to cause a nut 1808 to move in the vertical direction. The nut is attached to a sliding tube 1807, which is in contact with the instrument’s arm 1215, and causes the motor-end of the arm 1215 to also move vertically. A spring biases motor-end of the arm 1215 downwards - i.e. away from the direction of dispensing, so that when the direction of the motor 1610 reverses and the sliding tube 1807 moves downwards, the arm 1215 remains in contact with the sliding tube 1807 and therefore also moves in a downward direction. The cartridge actuator motor 1610 continues to alternate the rotation of the leadscrew 1809 so that the instrument arm 1215 performs small vertical mixing movements as indicated by the arrow 1810.

[0141]

[0145] In some embodiments, to performing the dispensing, the controller 1601 causes the cartridge actuator motor 1610 to turn the leadscrew 1809 in the direction to cause the nut 1808 and sliding tube 1807 to move to a maxim vertical position, casing the motor-end of the instrument’s arm 1215 to move to a maximum vertical position, and the cartridge-end of the arm 1215 to fully depress the dispense component 150, and perform the dispensing. The direction of this dispensing movement is indicated by the arrow 1811.

[0142]

[0146] FIG. 19 is an isometric view of a portion of the instrument 1500 including diagnostic test assembly, comprising a cartridge 1000 fitted with a dispense component 150. The 3D shape of the instrument arm 1215 is apparent, and although the lid 1520 is not shown, the position of the instrument arm 1215 within the dispense component 150 indicates that the lid 1520 is in the closed position.

[0143]

[0147] FIG. 20 is a flow diagram of a workflow 2000 of the instrument, according to some embodiments. At 2002, a user slides open the lid 1520 of the instrument, which can be latched in the open configuration. At 2004, the user inserts that cartridge 1000 into a cartridge holder of the instrument 1500. When the cartridge is received by the cartridge holder, the instrument detects the cartridge 1000, at 2006. In some embodiments, detection of the cartridge 1000 by the instrument 1500 causes the barcode reader to scan or read a barcode displayed on the cartridge 1000, allowing the instrument 1500 to identify the cartridge 1000, at 2008. For example, based on the identification of the cartridge, the instrument 1000 can determine which workflow from a set of workflows should be instigated. At 2010, the instrument 100 configures itself to perform the selected workflow, including setting suitable temperatures. For example, the instrument 1500 may cause the lysis chamber heater block 1410 and the reaction chamber heater block 1420 to operate a respective select temperatures.

[0144]

[0148] At 2012, the instrument 1500 prompts the user to remove the shipping closure or cap from the cartridge 1000 and to add the sample. At 2014, the user is then prompted to secure the dispense component 2150 to the cartridge 1000.

[0145]

[0149] In some embodiments, once the dispense component 2150 has been secured to the cartridge 1000 and is detected by the instrument 1500, the instrument 1500 causes the lid 1520 to close, at 2016. In some embodiments, once the dispense component 2150 has been secured to the cartridge 1000 and is detected by the instrument 1500, the user is prompted to close the lid 1520, at 2016. At this stage, no further user input is required to operate the instrument or to perform the test processes.

[0146]

[0150] At 2018, once the instrument 1500 detects that the lid 1520 is closed, the instrument 1500 causes a level mechanism, such as an arm of the instrument, to engage with the dispense component 2150 secured to the cartridge 1000.

[0151] At 2020, the mixture in the cartridge body is brought up to temperature for lysis and processing by the lysis chamber heater block 1410.

[0147]

[0152] At 2022, the instrument 1500 instigates the mixing function. For example, the instrument may cause the arm of the instrument to actuate the dispensing portion 207 to adopt the mixing state, and move longitudinally (for example, up and down) within the cartridge body to mix the mixture in the lysis chamber 480, without causing perforation of the membrane(s). The mixture may be maintained in the lysis chamber 480 for a required time, at a required temperature and multiple mixing cycles may be performed.

[0148]

[0153] At 2024, the instrument instigates the dispensing function. For example, the instrument 1500 causes the arm to actuate the dispensing portion 207 such that it adopts the dispensing state, causing the dispensing portion 207 to move longitudinally and protrude from the cartridge body to disrupt the membrane(s) and dispense the mixture into the reaction chamber. In some embodiments, the mixture, or fluid, is dispensed into the amplification tubes of the reaction chamber.

[0149]

[0154] At 2026, the instrument controls the temperature of the fluid in the amplification tubes, for example, by controlling the temperature of the reaction chamber heater block 1420.

[0150]

[0155] At 2028, the instrument 1500 acquires measurements from the fluid in the amplification tubes.

[0151]

[0156] At 2030, the instrument 1500 calculates the final test result or quantity based on the acquired measurements.

[0152]

[0157] At 2032, the instrument 1500 causes the display to indicate that the test is complete. In some embodiments, the instrument 1500 further displays the final test result or the quantity in combination with the indication that the test is complete.

[0153]

[0158] Described herein so far are two of many possible shipping closures for example, the cap 110 and foil seal 610, as well as three of many possible cartridge designs i.e. the clip together cartridge 100, the welded cartridge 500, the cartridge with a screw thread 600 and the cartridge with targeted dispensing 1000. As the skilled person will appreciate, there are also many possible configurations for the dispense component 150, which do not deviate from the spirit and scope of the present invention. Some example alternative designs are described below.

[0154]

[0159] FIG. 21 A shows an isometric view of a diagnostic test assembly comprising a cartridge 600 fitted with a dispense component 2150 in an undispensed state, FIG. 2 IB shows an isometric view of the diagnostic test assembly with the dispense component in a dispensed state, and FIG. 21C shows an expanded isometric view of the dispense component 2510, according to some embodiments.

[0155]

[0160] The dispense component 2150 comprises a securing portion 2110, and a dispensing portion 2113. In some embodiments, as illustrated, the securing portion 2110 comprises a cap. A cap seal 2135 and / or a plunger seal 2130 may be provided to improve securing and / or sealing function between the dispense component 2150 and a cartridge to which it is to be secured.

[0156]

[0161] In some embodiments, as illustrated, the dispensing portion 2113 may comprise a plunger head 2140 and a plunger body 250. The plunger body 250 may be substantially similar to that described above with reference to Figs. 2A, 2B and 3. For example, the plunger body 250 may comprises piercing tips 260 and may comprise piercing tip seals 270.

[0157]

[0162] In this example, the securing portion 2110 in the form of a cap defines an opening 2143 at an end 2146, for example, it’s free end. The opening 2143 of the securing portion 2110 is configured to receive an upper portion of the plunger head 2140 such that the upper portion 2142 of the plunger head 2140 extends through the securing portion 2110 when the securing portion 2110 is attached to the dispensing portion 2113. For example, the upper portion may be a substantially elongate member. When the securing portion 2110 is attached to the dispensing portion 2113 and the upper portion of the plunger head extends through the securing portion, the upper portion is actuatable to cause the dispensing portion to move relative to the securing portion.

[0158]

[0163] In this example, the securing portion 2110 comprises an inner thread (not shown) configured to cooperate with a complimentary thread on the cartridge to allow the dispense component 2150 to be secured to, or screwed onto, the cartridge 600. When the securing portion 2110 is being secured or threaded onto the cartridge, the securing portion 2110 rotates about the upper portion 2142 of the plunger head 2140. For example, the securing portion 2110 and upper portion 2142 are spun relative to the lower portion 2140 of the plunger head 2140 to connect the dispense component 2150 to the cartridge.

[0159]

[0164] The separate, spinning portion of the plunger head 2142 enables the cap 2110 to spin relative to the plunger head 2140 and body 250, which slide into the cartridge 600 without spinning as the cap is screwed on. To actuate a dispense function, a user or instrument presses down on the upper portion 2142 of the plunder head 2140 protruding through the cap in the direction of the arrow 2155. A full depressing of the upper portion 2142 causes the dispensing portion 2113 to move relative to and away from the securing portion 2110 to a first extent, causing the piercing tips 260 of the plunger body to disrupt the membrane(s) causing the dispensing function, as discussed above.

[0160]

[0165] FIG. 22Ais an isometric view of a diagnostic test assembly comprising a pneumatically actuated dispense component 2250 fitted on a cartridge 600, according to some embodiments and FIG. 22B is an isometric view of the dispensing portion 2207 (the dispense component 2250 with its securing portion 2210 removed). The securing portion 2210, in this example, is in the form of a cap which defines an opening 2215 at its free end 2213. The cap 2210 may be formed of a single part. The dispensing portion 2207 comprises chamber 2220 in fluid communication with the opening 2215 and a connected plunger body 250 comprising piercing tips 260. The opening 2215 of the cap provides access for an instrument pneumatic interface to apply pressure to the chamber 2220 of a dispensing portion 2207 of the dispense component 2250. Thus, the dispensing portion is configured to be pneumatically actuated by the application of air through the aperture to pressurise the chamber. In use, the instrument pumps air into the opening 2220 as indicated by arrow 2255, which pressurises the chamber 2220, causing the chamber 2220, and connected plunger body 250 with associated piercing tips 260 to move relative to and away from the securing portion 2210, for example, downwards within the cartridge body to which it attached, to actuate a dispense by disrupting membrane(s) as discussed above. In some embodiments, a gasket seal 2260 is provided to mitigate or prevent pressurisation of the lysis chamber 480.

[0161]

[0166] FIG. 23A is an isometric view of a diagnostic test assembly comprising a spring- actuated dispense component 2350 fitted on a portion of a cartridge 600 with an upper section 2316 of the securing portion in place , FIG. 23B is an isometric view of the diagnostic test assembly with the upper section 2316 of the securing portion removed and in the undispensed state, and FIG. 23C is an isometric view of the diagnostic test assembly with the upper section 2316 of the securing portion removed and in the dispensed state. The securing portion of the dispense component 2350 comprises a lower or base portion 2317, which houses a sliding bracket 2340. For example, and as shown., the sliding bracket 2340 may have an elongate (for example, pear-shaped) opening 2320 with a relatively narrow end or portion 2321 and a relatively broader or wider portion or end 2322. The dispense component 2350 further comprises a plunger rod 2370 connected to a spring (not shown) and attached to a plunger body 250 (not shown). The spring is biased towards the plunger body. The plunger body 250 and associated piercing tips 260, piercing tip seals 270 etc. are equivalent to those of the previously described dispense component 150.

[0162]

[0167] An end of the rod 2370 is configured to be received by the elongate opening 2320. When the sliding bracket 2340 is configured to transition, by way of a sliding motion, between an unactuated state, wherein the end of the rod is retained within the relatively narrow end 2321 of the elongate opening 2320, and an actuated state, wherein the end of the rod 2370 is released through the relatively broad end 2322 of the of elongate opening 2330, and is drawn away from the securing portion by action of the spring.

[0163]

[0168] The plunger rod 2370 may have a protruding rim 2371. In some embodiments, in the undispensed state, the plunger rod 2370 is held in place by an interaction between the protruding rim 2371 and the narrow portion 2321 of the opening 2320 of the sliding bracket 2340, as shown in FIG. 23B. When the sliding bracket is moved in the direction of the arrow 2355, the wider part 1222 of the opening 1220 slides towards the rod 1270. The wider portion 1222 of the opening 1220 is wide enough to enable the rod 1270 to slide through the opening. The spring (not shown) then drives the plunger body 250 further into the cartridge 100, causing the piercing tips 260 of the plunger body 250 to disrupt the membrane(s) and dispensing occurs, as shown in FIG. 23C.When fitted, the upper section 2316 protects the sliding bracket 2340 from accidental movement during storage or shipping. The upper section 2316 may have a small hole 2315 disposed in a wall of the upper section, through which an actuator pin of an instrument or a human operator can be inserted to cause the dispense component 2350 to actuate.

Claims

1. CLAIMS1. A diagnostic test assembly for use in conducting biological or environmental tests, the diagnostic test assembly comprising: a cartridge comprising: a cartridge body configured to house sample preparation fluid and configured to receive a biological or environmental sample through an opening; a reaction chamber comprising one or more amplification tubes; and one or more membranes configured to prevent fluid communication between the cartridge body and the one or more amplification tubes of the reaction chamber; and a dispense component configured to cooperate with the cartridge, wherein the dispense component comprises: a securing portion configured to secure the dispense component to the cartridge; and a dispensing portion configured to extend within the cartridge body of the cartridge when the dispense component is secured to the cartridge, and in response to actuation, the dispensing portion is configured to move relative to the securing portion; wherein, when the dispense component is secured to the cartridge and the dispensing portion is selectively actuated to a first extent, the dispensing portion moves a first distance within the cartridge body, away from the securing portion toward the reaction chamber, and disrupts at least one of the one or more membranes, bringing the cartridge body into fluid communication with at least one of the one or more amplification tubes of the reaction chamber, and allowing fluid within the cartridge body to be dispensed from the cartridge body into the at least one of the one or more amplification tubes.

2. The diagnostic test assembly of claim 1, wherein the dispensing portion is substantially elongate and is configured to move longitudinally relative to the securing portion.

3. The diagnostic test assembly of any one or the preceding claims, wherein securing the securing portion to the cartridge body does not cause actuation of the dispensing portion.

4. The diagnostic test assembly of any one or the preceding claims, wherein, when the dispense component is fitted to the cartridge and the dispensing portion is selectively actuated to a second extent, the dispensing portion moves a second distance within the cartridge bodyaway from the securing portion and without disrupting the one or more membranes to agitate fluid within the cartridge body, wherein the second distance is less than the first distance.

5. The diagnostic test assembly of claim 4, wherein selective actuation of the dispensing portion to the second extent followed by return of the dispensing portion toward the securing portion causes the dispensing portion to perform a reciprocal motion to thereby mix fluid within the cartridge body.

6. The diagnostic test assembly of any one of the preceding claims, wherein the dispensing portion comprises one or more piercing tips to disrupt the at least one of the one or more membranes, and wherein in response to actuation of the dispensing portion to the first extent, the one or more piercing tips of the dispensing portion protrude from the cartridge body and disrupt the at least one of the one or more membranes.

7. The diagnostic test assembly of claim 6, wherein the dispensing portion comprises a plunger head connected to a plunger body, and wherein the one or more piercing tips protrude from the plunger body and wherein the plunger head is configured to be actuated to cause the dispensing portion to move relative to the securing portion.

8. The diagnostic test assembly of claim 7, wherein the plunger body comprises one or more leg members, each leg member corresponding to a respective one of the one or more amplification tubes, and each leg member comprising a respective one of the one or more piercing tips, and wherein each of the one or more piercing tips is equipped with a tip seal to facilitate movement of the one or more piercing tips relative to internal walls of the amplification tubes when the dispense portion moves relative to the securing portion.

9. The diagnostic test assembly of claim 7 or claim 8, wherein the plunger head defines a first aperture in a side wall of the plunger head, the first aperture being arranged to receive an actuating member, which when it exerts a force, causes the dispensing portion to move relative to the securing portion.

10. The diagnostic test assembly of any one of the preceding claims, wherein the securing portion comprises a cap defining a chamber arranged to receive an upper section of thedispensing portion, wherein the cap defines a second aperture to provide access to the chamber, and wherein actuation of the dispensing portion can be performed through the second aperture.

11. The diagnostic test assembly of claim 10 when dependent on claim 9, wherein the first and second apertures cooperate with one another such that both the first and second apertures are arranged to receive the actuating member therethrough to allow for actuation of the dispensing portion.

12. The diagnostic test assembly of any one of claims 1 to 3 or any one of claims 6 to 8 when dependent on any one of claims 1 to 3, wherein an end of the securing portion defines an opening configured to receive an upper portion of the plunger body such that the upper portion of the plunger head extends through the securing portion and is actuatable to cause the dispensing portion to move relative to the securing portion when the securing portion is attached to the dispensing portion.

13. The diagnostic test assembly of any one of claims 1 to 3 or any one of claims 6 to 8 when dependent on any one of claims 1 to 3, wherein the dispensing component is pneumatically actuated.

14. The diagnostic test assembly of claim 13, wherein the dispensing portion comprises a chamber connected to a plunger body, wherein an end of the securing portion defines an opening in fluid communication with the chamber of the dispensing portion, and where the dispensing portion is configured to be pneumatically actuated by the application of air through the aperture to pressurise the chamber.

15. The diagnostic test assembly of any one of claims 1 to 3 or any one of claims 6 to 8 when dependent on any one of claims 1 to 3, wherein the dispensing component is spring actuated.

16. The diagnostic test assembly of claim 15, wherein the securing portion comprises base portion housing a sliding bracket defining an elongate opening, wherein the elongate opening is relatively broad at one end and relatively narrow at another end, and wherein the dispensing portion comprises a rod connected to a spring and a plunger body, where the spring is biased towards the plunger body, and wherein an end of the rod is configured to be received by theelongate opening, and wherein when the sliding bracket is configured to transition, by way of a sliding motion, between an unactuated state, wherein the end of the rod is retained within the relatively narrow end of the elongate opening, and an actuated state, wherein the end of the rod is released through the relatively broad end of the of elongate opening, and is drawn away from the securing portion by action of the spring.

17. The diagnostic test assembly of any one of the preceding claims, wherein the securing portion comprising a screw thread configured to engage with a complimentary screw thread disposed on the cartridge body to thereby secure the dispense component to the cartridge.

18. The diagnostic test assembly of any one of claims 1 to 16, wherein the securing portion comprising one or more clips and / or protrusions configured to engage with complimentary clips and / or protrusions disposed on the cartridge body to thereby secure the dispense component to the cartridge.

19. The diagnostic test assembly of any one of the preceding claims, wherein the reaction chamber comprises one or more nozzles to improve placement of dispensed fluid in the one or more amplification tubes.

20. The diagnostic test assembly of any one or the preceding claims, wherein the dispensing portion is configured to be selectively actuated by a cooperating arm of an instrument configured to conduct the biological or environmental test using the diagnostic test assembly.

21. The diagnostic test assembly of any one or the preceding claims, wherein the dispensing portion is configured to be selectively actuated by an actuating member.

22. The diagnostic test assembly of any one of the preceding claims, further comprising a removable closure, such as a cap or foil closure, configured to close the opening of the cartridge body for storage and / or shipping of the cartridge.

23. The diagnostic test assembly of any one of the preceding claims, wherein:(i) the cartridge body and the reaction chamber are welded together to form the cartridge;(ii) the cartridge body and the reaction chamber each comprise complementary components to allow the cartridge body and the reaction chamber to clip together to form the cartridge; or(iii) the cartridge body and the reaction chamber each comprise complementary thread to allow the cartridge body and the reaction chamber to screw together to form the cartridge.

24. The diagnostic test assembly of any one of the preceding claims wherein at least one of the one or more amplification tubes houses preloaded dry or lyophilised reagents which are configured to dissolve when fluid is dispensed into the one or more amplification tubes.

25. The diagnostic test assembly of any one of the preceding claims, wherein at least one of the one or more of the amplification tubes houses at least a preloaded magnetic mixing bead, which can be agitated by movement of one or more permanent magnets near the reaction chamber to mix content of the amplification tubes.

26. The diagnostic test assembly of any one of the preceding claims, wherein the cartridge body comprises the sample preparation fluid.

27. The diagnostic test assembly of any one of the preceding claims, wherein the cartridge body and / or the reaction chamber and / or the one or more amplification tubes are partially or entirely composed of a transparent material such that the contents thereof, and reactions that take place therein, can be observed visually or optically sensed by an instrument.

28. The diagnostic test assembly of any one of the preceding claims, wherein the one or more membranes comprises:(i) a foil seal which covers all the one or more amplification tubes of the reaction chamber; or(ii) one or more foil seals, each of the one of more foil seals configured to cover a respective one of the one or more amplification tubes of the reaction chamber.

29. An instrument configured to perform a biological or environmental test using the diagnostic test assembly of any one of claims 1 to 28, the instrument comprising:a controller including one or more processors and memory, wherein memory comprises computer executable instructions for performing the biological or environmental test; a cartridge holder configured to receive the cartridge of the diagnostic test assembly; and an arm configured to selectively engage with the dispense component of the diagnostic test assembly to cause the dispensing portion of the dispense component to perform a mixing or dispensing function under the control of the controller.

30. A method of performing a biological or environmental test using the diagnostic test assembly of any one of claims 1 to 28.

31. The method of claim 30, further comprising: providing a biological or environmental sample to the cartridge body, wherein the cartridge body comprises a sample preparation fluid; securing the dispense component to the cartridge; controlling, by the instrument, a temperature of the mixture in the cartridge body for lysis; causing an arm of an instrument configured to perform a biological or environmental test to actuate the dispensing portion to a first extent to cause the dispensing portion to move the first distance within the cartridge body, away from the securing portion toward the reaction chamber, and to disrupt the at least one of the one or more membranes, bringing the cartridge body into fluid communication with at least one of the one or more amplification tubes of the reaction chamber, and allowing fluid within the cartridge body to be dispensed from the cartridge body into the at least one of the one or more amplification tubes.

32. The method of claim 31 when using the diagnostic test assembly of any one of claims 1 to 3 or 6 to 28 when dependent on any of claims 1 to 3, wherein controlling by the instrument, a temperature of the mixture in the cartridge body for lysis, further comprises: causing the arm of the instrument to actuate the dispense component to a second extent to cause the dispensing portion to move the second distance within the cartridge body away from the securing portion and without disrupting the one or more membranes to agitate fluid within the cartridge body.

Images