Sample processing cartridge and method for using the same

Abstract

The present disclosure provides cartridges, systems, and methods for introducing a sample into a chamber. The cartridges, systems, and methods of the present disclosure may be used for isolating an analyte from a sample.

Description

[0001] SAMPLE PROCESSING CARTRIDGE AND METHOD FOR USING THE SAME

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] Pursuant to 35 U.S.C. § 119(e), this application claims priority to the filing dates of United States Provisional Patent Application Serial No. 63 / 660,900 filed June 17, 2024, the disclosure of which application is herein incorporated by reference in its entirety.

[0004] INTRODUCTION

[0005] Analysis of a biological entity such as proteins or nucleic acids, in biological sample generally involves isolating the target entity from the mixture of non-target biological entities. The isolated target analyte is analyzed using downstream applications, such as, amplification of a nucleic acid for detection and quantification, immunoassay, and the like. Approaches that include column-based isolation and purification, reagent-based isolation and purification, magnetic beadbased isolation and purification, and other technologies are used to isolate target analytes. Reagents, kits and instruments that find use in isolating and purifying nucleic acids are available. For effective sample preparation, it is important that the isolated target analyte does not contain residues of certain reagents and / or solvents used during isolation. Poor sample preparation can lead to suboptimal results in downstream applications, and it is for this reason that optimized versions of kits have emerged to address variation in sample source, be it blood, plant tissue, fungi, bacteria, or virus.

[0006] Existing methods include multiple steps such as releasing a nucleic acid from its native biological source (e.g., lysis of cells, such as patient cells or lysis of microorganisms, such as, virus, bacteria, fungi, etc.) using chaotropic nucleic acid extraction technology, binding of nucleic acids to a solid phase (e.g., paramagnetic particles) using silica or iron oxide nucleic acid chemistry, separation of the solid phase from the residual lysis solution using magnetic separation technology, washing to remove unwanted materials, and elution or separation of nucleic acid from the solid phase using fluid handling technology. At the completion of the sample preparation protocol, the liquid comprising nucleic acid is transferred to a collection container(s) such as PCR tubes or strips. In such multistep processes, there is an increased possibility of contamination or even loss of sample. There is a need for systems and methods for effective isolation of target analyte. Also, exists an interest in automating all or individual aspects of sample preparation to increase throughput, decrease user error and / or limit exposure of users to harmful substances. SUMMARY

[0007] The present disclosure provides cartridges, systems and methods for introducing a sample into a chamber. In certain aspects, the present disclosure provides cartridges, systems and methods for isolating an analyte from a sample. The cartridges, systems and methods of the present disclosure, for isolating an analyte from a sample may involve a plurality of chambers, a plurality of solution dispensing pouches and an actuator.

[0008] Aspects of the invention include cartridges for introducing a sample into a chamber. In certain aspects, the cartridge may be used for isolating an analyte from a sample. Cartridges of interest may comprise a plurality of chambers, each said chamber configured for holding a solution and comprising a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a channel connected to the inlet of each chamber.

[0009] The cartridges of interest may include a first chamber configured for filling with a first solution; a second chamber configured for filling with a second solution; and a third chamber configured for filling with a third solution. In some embodiments, the cartridges of interest include an additional chamber that is configured for filling with a wash buffer, wherein the additional chamber is positioned between the second chamber and the third chamber and is connected to the channel. In some embodiments, each said chamber comprises an opening on the second side of the chamber, wherein the opening is located adjacent to the top end or in the inlet of the chamber. In some embodiments, the cartridges of interest further comprise a plurality of solution dispensing pouches, each said solution dispensing pouch comprising a first layer and a second layer opposite to the first layer enclosing a solution, and the first layer is positioned immediately adjacent to the second side of the chamber. In some embodiments, the first layer of each solution dispensing pouch ruptures more easily as compared to the second layer. For example, the first layer of each solution dispensing pouch has a lower elongation at break than the second layer. Elongation at break, also known as fracture strain or tensile elongation at break, is a material's ability to stretch or deform before it breaks. The first layer may be manufactured such that it has a lower elongation at break as compared to the second layer and will fracture under a tensile load while the second layer deforms and does not fracture under the tensile load.

[0010] Aspects of the invention include systems for isolating an analyte from a sample. Systems of interest comprise the cartridge; and an actuator, wherein the first layer of pouch is positioned immediately adjacent the opening on the second side of the chamber and the second layer of the pouch is positioned adjacent the actuator, the actuator comprises a blunt member configured for applying pressure on the second layer of the pouch such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured, and the blunt member is aligned with the opening of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening.

[0011] In some embodiments, each said chamber in the cartridge comprises two protrusions extending towards the pouch and flanking the opening on the second side and configured to contact the first layer of the pouch and to accommodate the blunt member between the two protrusions when the actuator applies pressure to the pouch. When the blunt member applies pressure on the second side and through the pouch, the two protrusions pierce or tear the first side of the pouch but not the second side of the pouch because the second side of the pouch is either thicker or has a higher tear resistance. In some embodiments, the pouch comprises a rigid member positioned or positionable adjacent to the opening such that the blunt member contacts second layer and applies pressure to the first layer via the rigid member. In some embodiments, the rigid member is a magnetically responsive solid member. In some embodiments, the rigid member is a paramagnetic bead or ferromagnetic bead.

[0012] In certain aspects, systems of interest comprise the cartridge; and an actuator comprising a hollow member, wherein each said chamber comprises two protrusions extending towards the pouch and flanking the opening on the second side; the first layer of the pouch is positioned immediately adjacent the opening on the second side and the second layer of the pouch is positioned adjacent the actuator, the hollow member configured for applying pressure on the second layer of the pouch such that the first layer is stretched against the protrusions and ruptures, and the hollow member is aligned with the opening on the second side such that the first layer is ruptured at a point immediately adjacent to the opening.

[0013] Aspects of the invention include semi- automated or automated methods of using the system for introducing a sample into a chamber. In certain aspects, the present disclosure includes semi-automated or automated methods of using the system for isolating an analyte from a sample. In some embodiments, methods of interest comprise activating the actuator to apply pressure on the pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the solution from the pouch enters the chamber via the opening.

[0014] Aspects of the invention include semi-automated or automated methods for introducing a sample into a chamber. In certain aspects, the present disclosure includes semi- automated or automated methods for isolating an analyte from a sample. In some embodiments, methods of interest comprise providing the system; introducing the sample into the first chamber; activating the actuator to apply pressure on the pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the first solution from the first pouch enters the first chamber via the first opening area; mixing the sample and first solution in the first chamber; wherein the first chamber comprises magnetic beads configured for binding to the analyte; moving the magnetic beads to the second chamber; moving the actuator to a position adjacent the second pouch; activating the actuator to apply pressure on the second pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the second solution from the second pouch enters the second chamber via the second opening; moving the magnetic beads to the third chamber; moving the actuator to a position adjacent the third pouch; activating the actuator to apply pressure on the third pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the third solution from the third pouch enters the third chamber via the third opening; draining the third solution from the third chamber into the detection wells via the channel connecting the third chamber to the detection wells, wherein the drained third solution comprises the isolated analyte when the analyte is present in the sample; and detecting the isolated analyte in the detection wells.

[0015] BRIEF DESCRIPTION OF THE FIGURES

[0016] FIG. 1A depicts a view of the cartridge for isolating an analyte from a sample in which view the chambers, the channel connecting the chambers and detection wells are visible.

[0017] FIG. IB depicts a view of the cartridge for isolating an analyte from a sample in which view pouches that are associated with the chambers are visible.

[0018] FIG. 2 depicts a cartridge for isolating an analyte from a sample. The cartridge includes three chambers connected via a channel and a pouch associated with each chamber.

[0019] FIG. 3A depicts the front view of a cartridge for isolating an analyte from a sample.

[0020] FIG. 3B depicts the front view of a cartridge for isolating an analyte from a sample.

[0021] FIG. 4 depicts a schematic of a system for piercing a first layer of a pouch without piercing the second layer of the pouch. The actuator (8) comprises a blunt member (9) configured for applying pressure on the second layer (6) of the pouch such that the second layer is pushed into the first layer (5) causing the first layer to stretch and rupture and the second layer is not ruptured, and the blunt member is aligned with the opening (3) of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening.

[0022] FIG. 5 depicts a schematic of a system for piercing a first layer of a pouch without piercing the second layer of the pouch. In this embodiment, the chamber in the cartridge comprises two protrusions extending towards the pouch and flanking the opening on the second side and configured to contact the first layer of the pouch and to accommodate the blunt member between the two protrusions when the actuator applies pressure to the pouch. FIG. 6 depicts a schematic of a system for piercing a first layer of a pouch without piercing the second layer of the pouch. In this embodiment, the actuator comprises a hollow member configured for applying pressure on the second layer of the pouch such that the first layer is stretched against the protrusions and ruptures, and the hollow member is aligned with the opening on the second side such that the first layer is ruptured at a point immediately adjacent to the opening.

[0023] DETAILED DESCRIPTION

[0024] Aspects of the present disclosure include a cartridge, a system and a method for introducing a sample into a chamber. The cartridge, system and method may be useful for isolating an analyte from a sample.

[0025] Before the present devices, systems, and methods are described in greater detail, it is to be understood that the present disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0026] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the present systems, devices and methods. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the systems, devices and methods, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the systems, devices and methods.

[0027] Certain ranges are presented herein with numerical values being preceded by the term “about.” The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating un-recited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number.

[0028] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present systems, devices and methods, representative illustrative systems, devices and methods are now described. The present disclosure may be understood more readily by reference to the following detailed description of desired embodiments and the examples included therein. In the following specification and the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings.

[0029] The term “comprising” is used herein as requiring the presence of the named component and allowing the presence of other components. The term “comprising” should be construed to include the term “consisting essentially of’ and “consisting of.” The “consisting essentially of’ allows the presence of the named component(s), along with other components which do not change the function / structure of the named component(s). The “consisting of’ allows the presence of the named component(s), along with any adhesives or other bonding means for attaching the listed component(s).

[0030] Numerical values should be understood to include numerical values which are the same when reduced to the same number of significant figures and numerical values which differ from the stated value by less than the experimental error of conventional measurement technique of the type described in the present application to determine the value.

[0031] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 2 ml to 10 ml” is inclusive of the endpoints, 2 ml and 10 ml, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and / or values.

[0032] The modifier “about” used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context. When used in the context of a range, the modifier “about” should also be considered as disclosing the range defined by the absolute values of the two endpoints. For example, the range of from about “2 to about 10” also discloses the range “from 2 to 10.” The term “about” may refer to plus or minus 10% of the indicated number. For example, “about 10%” may indicate a range of 9% to 11%, and “about 1” may mean from 0.9- 1.1.

[0033] It should be noted that many of the terms used herein are relative terms. For example, the terms “top” and “bottom” are relative to each other in location and refer to surfaces where the top is always higher than the bottom relative to an absolute reference, i.e., the surface of the earth. The terms “inlet” and “outlet” are relative to a fluid flowing through them with respect to a given structure, e.g., a fluid flows through the inlet into the structure and flows out of the structure through the outlet. The terms “upwards” and “downwards” are also relative to an absolute reference; upwards is always against the gravity of the earth while downwards is always towards the gravity of the earth.

[0034] The term “parallel” should be construed in its lay sense of two surfaces that maintain a generally constant distance between them, and not in the strict mathematical sense that such surfaces will never intersect when extended to infinity.

[0035] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0036] It is noted that, as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements or use of a “negative” limitation.

[0037] A “liquid reagent”, as the term is used herein, refers to any liquid contained within any of the chambers of the device as described herein, including aqueous, nonaqueous, and water- immiscible liquids.

[0038] A “reagent solution” typically refers to an aqueous solution. The “reagent” may be a chemical or biological substance that causes a chemical change to a sample component, or it may be simply a buffering agent, a salt, or a solvent.

[0039] As pertains to the present disclosure, a “biological sample” can include a tissue sample or a body fluid sample, which includes liquid, solid, and semisolid samples, e.g. blood, blood components such as plasma or serum, urine, saliva, sputum, mucous, amniotic fluid, semen, vaginal secretions, tears, spinal fluid, washings, feces, biopsy specimens, skin, nails, and hair.

[0040] A “specific binding member” or “affinity reagent”, as used herein, is a molecule or moiety that specifically binds to a target analyte through chemical or physical means. Immunoreactive specific binding members include antigens or antigen fragments and antibodies or functional antibody fragments. Other specific binding pairs include biotin and avidin, carbohydrates and lectins, complementary nucleotide sequences, effector and receptor molecules, cofactors and enzymes, enzyme inhibitors and enzymes, and the like. In the extraction / isolation procedures described herein, a binding member is attached to a solid phase support, such as a plurality of paramagnetic particles, in order to extract the analyte from a sample containing non-target components. Following isolation of the particle-analyte complex from the non-target components, the complex is treated to effect removal of the analyte from the particles. Removal may be effected by, for example, heating the solution containing the complex and / or changing the chemical environment (e.g. salt concentration, pH, etc.). In other embodiments, a chemical or enzymatic reagent is used to disrupt the particle- analyte complex and thus effect removal of the analyte from the particles.

[0041] An “isolated” analyte is one that has been separated from other constituents with which it is associated in a sample, such that it can be detected with a desired degree of accuracy and precision. The isolated analyte is typically dissolved in a solvent medium that may also contain non-interfering substances. In the case of a biological sample, the analyte is isolated from cellular constituents with which it is normally associated, and from other types of cells which may be present in the sample.

[0042] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present device, systems and methods. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0043] CARTRIDGES

[0044] As summarized above, aspects of the present disclosure include a cartridge comprising a plurality of chamber. The cartridge may be used for isolating an analyte from a sample, such as biological sample. The analyte could be, as described further below, a protein, a nucleic acid, a virus, a cell or cell component.

[0045] The cartridge may be used for isolating an analyte manually, in an automated or semiautomated manner. For automated or semi- automated manner, the cartridge can be used in conjunction with an instrument such as described further below.

[0046] In certain aspects, the cartridge comprises a plurality of chambers, each said chamber configured for holding a solution and comprising a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a channel connected to the inlet of each chamber. In some embodiments, the cartridge comprises a first chamber configured for filling with a first solution; a second chamber configured for filling with a second solution; and a third chamber configured for filling with a third solution.

[0047] FIG. 1A represents an embodiment of a cartridge for introducing a sample into a chamber. The said cartridge may be used for isolating an analyte from a sample. This embodiment of a cartridge includes a plurality of chambers. For example, the cartridge includes a first chamber (201 ), a second chamber (202), a third chamber (203), and a fourth chamber (204). The first chamber is configured for filling with a first solution. The second chamber is configured for filling with a second solution. The third chamber is configured for filling with a third solution. The fourth chamber is configured for filling with a fourth solution. Each of the said chambers have an inlet at the top end and the inlets of the first chamber (201), the second chamber (202), the third chamber (203), and the fourth chamber (204), are connected via a channel (13). The inlets of each said chamber are located at substantially the same height with reference to the channel. The fourth chamber (204) comprises an opening (15) at the bottom end of the chamber, wherein the opening (15) is configured for draining the third chamber. The opening (15) of the fourth chamber (204) is connected to a plurality of detection wells (17) via a channel (16).

[0048] Each of the first, second and third chambers are configured to contain a water-miscible liquid reagent. For example, the first chamber may contain a reagent capable of cell and / or viral lysis, the second chamber may contain an aqueous wash solution, and the third chamber may contain an elution buffer. In some embodiments, the first solution is a lysis buffer; the second solution is a wash buffer; and the third solution is an elution buffer.

[0049] In some embodiments, the inlet of the first chamber is positioned lower than the inlet of the other chambers present in the cartridge, e.g., the second and third chambers. In some embodiments, the inlets of the plurality of chambers are located at substantially the same height with reference to the channel.

[0050] In some embodiments, the channel is substantially parallel to the top ends of the chambers.

[0051] The chambers may have any suitable shape and size. In some embodiments, the first chamber has a substantially trapezoidal shape. In some embodiments, the second chamber has a substantially square shape. In some embodiments, the third chamber has a substantially pentagonal shape. The chambers may include baffles located at the top end. For example, the baffles may be curved walls or curved ceiling that redirects the upward moving solution during mixing into a downward trajectory back into the primary volume of solution in that chamber. The baffles prevent or minimize solutions from jetting or spilling into the adjacent chamber during the mixing steps. For example, the top end of the chambers, e.g., the first chamber configured for mixing a sample with a lysis buffer may have a top end that curves downwards towards the inlet such that the height of the top end is lower at the inlet and higher in regions surrounding the inlet.

[0052] A cartridge of the present disclosure may be substantially rectangular in shape and the chambers may be substantially flat with a low depth relative to the width and height. The top end of the first chamber may curve downwards towards the inlet which may be centrally located at the top end.

[0053] Tn some embodiments, the first chamber further contains a plurality of solid carrier particles, which may comprise a plurality of paramagnetic particles. The cartridge may be supplied with the particles, or they may be added to the first chamber prior to or during use.

[0054] In certain embodiments, the particles are magnetic particles. One or more of the paramagnetic particles is typically treated on its surface with an affinity reagent capable of associating with an analyte; the affinity reagent may be, for example, an antibody or antibody fragment with specific binding for an analyte, such as a protein, or a nucleic acid sequence capable of hybridizing with an analyte. The paramagnetic particles may be present in an inlet of the chamber.

[0055] In some embodiments, the third chamber comprises an opening at a bottom end of the chamber, wherein the opening is configured for draining the third chamber. In some embodiments, the third chamber is connected to a plurality of detection wells via a channel.

[0056] In some embodiments, the cartridge comprises an additional chamber, wherein the additional chamber is configured for filling with a wash buffer, wherein the additional chamber is positioned between the second chamber and the third chamber and is connected to the channel. The cartridge may include further chambers in addition to described above, and in addition to those illustrated.

[0057] In some embodiments, each said chamber comprises an opening on the second side of the chamber, wherein the opening is located adjacent to the top end or in the inlet of the chamber.

[0058] In certain aspects, each chamber has a length and a width, wherein the length is greater than the width. The length may be at least 1.5 times or 2 times greater than the width. The aspect ratio of the chambers may be determined by the fluid properties of the solution and it’s volume along with the displacement or amplitude of the mixing cycles. For example, the length of the chambers is about 2 times greater (e.g. 2-3 times greater) than the amplitude of the mixing.

[0059] In some embodiments, the cartridge further comprises a plurality of solution dispensing pouches, each said solution dispensing pouch comprising a first layer and a second layer opposite to the first layer enclosing a solution, and the first layer is positioned immediately adjacent to the second side of the chamber. In some embodiments, the cartridge further comprises a first pouch positioned adjacent to the first chamber; a second pouch positioned adjacent to the second chamber; and a third pouch positioned adjacent to the third chamber.

[0060] FIG. IB shows a cartridge comprising plurality of solution dispensing pouches, wherein each said solution dispensing pouch is positioned adjacent to each said chamber. For example, a first pouch (401) positioned adjacent to the first chamber (201), a second pouch (402) positioned adjacent to the second chamber (202), a third pouch (403) positioned adjacent to the third chamber (203), and a fourth pouch (404) positioned adjacent to the fourth chamber (204).

[0061] In some embodiments, the first layer of each solution dispensing pouch has a lower tensile strength than the second layer.

[0062] In some embodiments, the first layer and the second layer are sealed together to form the pouch.

[0063] In some embodiments, the first layer of the pouch is a thin aluminum foil, and the second layer is a thick aluminum foil. In some embodiments, the first layer and the second layer of the pouch are each a composite of different materials. For example, the first layer is a composite of two materials and the second layer is a composite of three materials. The materials may be aluminum, thermoplastic, a polymer, or a combination of one, two or all three materials. A composite layer may be formed by adhering layers of the different materials. The first layer may be a composite of an aluminum sheet coated with a thermoplastic adhesive and the second layer may be a composite of an aluminum sheet coated with a thermoplastic adhesive on one surface and coated with a polymer on the other surface. The first layer of the pouch may be a composite sheet of aluminum with a thermoplastic adhesion layer on the side facing the second layer. On the side facing the second side of the chamber on the cartridge, the aluminum may be bare and uncoated. The second layer may also be a composite layer with aluminum and an adhesion layer facing the first layer and a thicker polymer layer on the side facing the actuator. The elongation at break of the first layer may be at least 0.5 times (e.g., 0.6 times, 0.7 times, or 0.8 times) the elongation at break of the second layer.

[0064] In some embodiments, the cartridge further comprises an additional pouch associated with an additional chamber, wherein the additional pouch is positioned between the second pouch and the third pouch adjacent to the additional chamber positioned between the second and third chambers.

[0065] In certain aspects, the first layer of a solution dispensing pouch has a lower elongation at break than the second layer. For example, the first layer may rupture at 2%-30% elongation and the second layer may rupture at 50%-100% elongation; the first layer may rupture at 2%-20% elongation and the second layer may rupture at 60%-100% elongation; the first layer may rupture at 2%- 10% elongation and the second layer may rupture at 70%-100% elongation; or the first layer may rupture at 2%-5% elongation and the second layer may rupture at 80%-100% elongation. For example, the first layer may rupture at 2% elongation and the second layer may rupture at 90-100% elongation.

[0066] In certain aspects, the first pouch may hold liquid having a volume the range of about 3ML to 4ML. In certain aspects, the second pouch may hold liquid having a volume in the range of about 0.5ML to 1ML. In certain aspects, the third pouch may hold liquid having a volume in the range of about 0.5ML to 1ML.

[0067] In certain aspects, each pouch has a length and a width, wherein the length is greater than the width. The length may be at least 1.5 times or 2 times greater than the width. In certain aspects, the first pouch may have a length in the range of about 65MM to 75MM, and a width in the range of about 45MM to 55MM.

[0068] In certain aspects, the second pouch may have a length in the range of about 65MM to 75MM, and a width in the range of about 20MM to 30MM. In certain aspects, the third pouch may have a length in the range of about 65MM to 75MM, and a width in the range of about 20MM to 30MM.

[0069] FIG. 2 represents an example of a system for introducing a sample into a chamber. The said system may be used for isolating an analyte from a sample as per one of the embodiments of the present disclosure. This example of the system (1) includes a cartridge comprising a first chamber (201) having a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a first opening (301) on the second side of the chamber located adjacent to the top end or in the inlet of the chamber. The cartridge further comprises a second chamber (202) having a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a second opening (302) on the second side of the chamber located adjacent to the top end or in the inlet of the chamber. The cartridge further comprises a third chamber (203) having a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a third opening (303) on the second side of the chamber located adjacent to the top end or in the inlet of the chamber. The inlet of the first chamber (201) is positioned lower than the inlet of the second (202) and third chambers (203). The system comprises a channel (13) connected to the inlet of each chamber. The system further includes a first pouch (401) comprising a first layer and a second layer opposite to the first layer enclosing a solution, wherein the first layer has a lower tensile strength than the second layer, and the first layer is positioned immediately adjacent the first opening (301 ) on the second side of the chamber. The system further includes a second pouch (402) comprising a first layer and a second layer opposite to the first layer enclosing a solution, wherein the first layer has a lower tensile strength than the second layer, and the first layer is positioned immediately adjacent the second opening (302) on the second side of the chamber. The system further includes a third pouch (403) comprising a first layer and a second layer opposite to the first layer enclosing a solution, wherein the first layer has a lower tensile strength than the second layer, and the first layer is positioned immediately adjacent the third opening (303) on the second side of the chamber.

[0070] In some aspects, the cartridge may be provided in a system of components, where the components include solution dispensing pouches and an actuator. Examples of solution dispensing pouches are provided below.

[0071] The cartridge can be designed for automated or semi-automated use within a system that may hold one or a plurality of such cartridges, as described further below. Accordingly, the cartridge may contain external features, such as notches or ridges, used to properly align the cartridge within the system.

[0072] SYSTEMS

[0073] As summarized above, aspects of the present disclosure include a first system for introducing a sample into a chamber. In certain aspects, the present disclosure includes a first system for isolating an analyte from a sample.

[0074] In certain aspects, the system comprises the cartridge; and an actuator, wherein the first layer of the pouch is positioned immediately adjacent the opening on the second side of the chamber and the second layer of the pouch is positioned adjacent the actuator, the actuator comprises a blunt member configured for applying pressure on the second layer of the pouch such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured, and the blunt member is aligned with the opening of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening.

[0075] In the embodiment shown in Fig. 2, the first pouch (401) is filled with a first solution, the second pouch (402) is filled with a second solution, and the third pouch (403) is filled with a third solution. The first chamber (201) comprises a sample introduction region (12) for introducing a sample into the chamber (201). The channel (13) connects the inlet of first chamber (201), the inlet of second chamber (202) and the inlet of third chamber (203) for fluid communication. A sample is introduced into the first chamber (201) via the sample introduction region (12). An actuator is positioned adjacent to the first pouch (401) and applies pressure on the second layer of the first pouch (401) such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured. Upon rupture of the first layer, the first solution from the first pouch (401) enters the first chamber (201) via the first opening area (301). The sample then mixes with the first solution in the first chamber (201).

[0076] The system may comprise a magnetic bead (14) for binding to the analyte. The magnetic bead (14) in the first chamber (201) binds to the analyte. The magnetic bead (14) then moves to the second chamber (202) travelling via the channel ( 1 ) connecting the inlet of first chamber (201) to the inlet of second chamber (202). The magnetic bead may be able to pass through the chambers and channel upon application of an external force. In some embodiments, the external force is a magnetic force. Further, the actuator moves to a position adjacent the second pouch (402) and applies pressure on the second pouch (402) such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured. Upon rupture of the first layer, the second solution from second pouch (402) enters the second chamber (202) via the second opening area (302). The magnetic bead (14) then moves to the third chamber (203) travelling via the channel (13) connecting the inlet of second chamber (202) to the inlet of third chamber (203). The actuator then moves to a position adjacent the third pouch (403) and applies pressure on the third pouch (403) such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured. Upon rupture of the first layer, the third solution from the third pouch (403) enters the third chamber (203) via the third opening area (303). The third chamber (203) includes an opening (15) at the bottom end of the chamber for draining the third solution from the third chamber (203).

[0077] The system further comprises a channel (16) connecting the opening (15) of the third chamber (203) to a plurality of detection wells (17). The third solution is then drained into the detection wells (17) via the channel (16). The third solution comprises the isolated analyte which gets detected in detection wells (17).

[0078] FIG. 3A represents the front view of another example of a system for introducing a sample into a chamber. The said system may be used for isolating an analyte from a sample as per one of the embodiments of the present disclosure. This example of the system includes a cartridge comprising a first chamber (201) comprising a first opening (301) located into the inlet, a second chamber (202) comprising a second opening (302) located into the inlet, and a third chamber (203) comprising a third opening (303) located into the inlet. The system further comprises a channel (13) connecting the inlet of first chamber (201), the inlet of second chamber (202) and the inlet of third chamber (203) for fluid communication. In a preferred embodiment, each of the first, second and third chambers contain a water- miscible liquid reagent. For example, the first chamber may contain a reagent capable of cell lysis, the second chamber may contain an aqueous wash solution, and the third chamber may contain an elution medium. The additional chamber, if present between the second and third chamber, may contain a further wash solution.

[0079] In some embodiments, the first solution is a lysis buffer; the second solution is a wash buffer; and the third solution is an elution buffer.

[0080] In certain aspects, each chamber has a length and a width, wherein the length is greater than the width. The length may be at least 1.5 times or 2 times greater than the width.

[0081] In some embodiments, the cartridge further comprises a plurality of solution dispensing pouches, each said solution dispensing pouch comprising a first layer and a second layer opposite to the first layer enclosing a solution, and the first layer is positioned immediately adjacent to the second side of the chamber.

[0082] FIG. 3B represents the back view of the said system for introducing a sample into a chamber. The said system may be used for isolating an analyte from a sample as per one of the embodiments of the present disclosure. The system includes a first pouch (401) placed positioned adjacent to the first chamber (201), a second pouch (402) placed positioned adjacent to the second chamber (202), and a third pouch (403) placed positioned adjacent to the third chamber (203). In some embodiments, the solution in first pouch (401) is a first solution, the second pouch (402) is filled with a second solution, and the third pouch (403) is filled with a third solution. An actuator comprising a blunt member is positioned adjacent to the first pouch (401) such that the first layer of the pouch (401) is immediately adjacent to the first opening (301) on the second side of the chamber (201) and the second layer is adjacent the actuator. The actuator is aligned with the first opening (301) on the second side of the chamber (201). The blunt member of the actuator applies pressure on the second layer of the first pouch (401) such that the second layer is pushed into the first layer causing the first layer to stretch and rupture at a point (101) immediately adjacent to the opening (301). Further, the actuator moves to a position adjacent to the second pouch (402) and is aligned with the second opening (302). The blunt member of the actuator applies pressure on the second layer of the second pouch (402) such that the second layer is pushed into the first layer causing the first layer to stretch and rupture at a point (102) immediately adjacent to the opening (302). Further, the actuator moves to a position adjacent to the third pouch (403) and is aligned with the third opening (303). The blunt member of the actuator applies pressure on the second layer of the third pouch (403) such that the second layer is pushed into the first layer causing the first layer to stretch and rupture at a point (103) immediately adjacent to the opening (303). In some embodiments, the first layer of each solution dispensing pouch has a lower tensile strength than the second layer.

[0083] In some embodiments, the first layer and the second layer are sealed together to form the pouch.

[0084] In some embodiments, the first layer of the pouch is a thin aluminum foil, and the second layer is a thick aluminum foil.

[0085] In certain aspects, the first layer of a solution dispensing pouch has a lower tensile strength than the second layer. Preferably, the first layer has a tensile strength lower than 1.5 times that of the second layer. Preferably, the first layer has a tensile strength lower than 2 times that of the second layer. Preferably, the first layer has a tensile strength lower than 2.5 times that of the second layer.

[0086] Preferably, the first layer has a tensile strength in its longitudinal direction that is lower than 1.5 times that of the tensile strength in its longitudinal direction of the second layer. Preferably, the first layer has a tensile strength in its longitudinal direction that is lower than 2 times that of the tensile strength in its longitudinal direction of the second layer. Preferably, the first layer has a tensile strength in its longitudinal direction that is lower than 2.5 times that of the tensile strength in its longitudinal direction of the second layer.

[0087] Preferably, the first layer has a tensile strength in its transverse direction that is lower than 1.5 times that of the tensile strength in its transverse direction of the second layer. Preferably, the first layer has a tensile strength in its transverse direction that is lower than 2 times that of the tensile strength in its transverse direction of the second layer. Preferably, the first layer has a tensile strength in its transverse direction that is lower than 2.5 times that of the tensile strength in its transverse direction of the second layer. In certain aspects, the first layer of a solution dispensing pouch has a tensile strength in the range of about 45 MPa to about 55 MPa. Preferably, the first layer has a tensile strength in the range of about 49 MPa to about 52 MPa.

[0088] In certain aspects, the first layer of a solution dispensing pouch has a tensile strength in its longitudinal direction in the range of about 45 MPa to about 55 MPa. Preferably, the first layer has a tensile strength in its longitudinal direction in the range of about 49 MPa to about 52 MPa.

[0089] In certain aspects, the first layer of a solution dispensing pouch has a tensile strength in its transverse direction in the range of about 45 MPa to about 55 MPa. Preferably, the first layer has a tensile strength in its transverse direction in the range of about 49 MPa to about 52 MPa.

[0090] In certain aspects, the second layer of a solution dispensing pouch has a tensile strength in the range of about 95 MPa to about 120 MPa. Preferably, the second layer has a tensile strength in the range of about 100 MPa to about 110 MPa. In certain aspects, the second layer of a solution dispensing pouch has a tensile strength in its longitudinal direction in the range of about 95 MPa to about 120 MPa. Preferably, the second layer has a tensile strength in its longitudinal direction in the range of about 100 MPa to about 110 MPa.

[0091] In certain aspects, the second layer of a solution dispensing pouch has a tensile strength in its transverse direction in the range of about 95 MPa to about 120 MPa. Preferably, the second layer has a tensile strength in its transverse direction in the range of about 100 MPa to about 110 MPa.

[0092] In certain aspects, the first pouch may hold liquid in the range of about 3ML to 4ML. In certain aspects, the second pouch may hold liquid in the range of about 0.5ML to 1ML. In certain aspects, the third pouch may hold liquid in the range of about 0.5ML to 1ML.

[0093] In certain aspects, each pouch has a length and a width, wherein the length is greater than the width. The length may be at least 1.5 times or 2 times greater than the width. In certain aspects, the first pouch may have a length in the range of about 65MM to 75MM, and a width in the range of about 45MM to 55MM.

[0094] In certain aspects, the second pouch may have a length in the range of about 65MM to 75MM, and a width in the range of about 20MM to 30MM. In certain aspects, the third pouch may have a length in the range of about 65MM to 75MM, and a width in the range of about 20MM to 30MM.

[0095] FIG. 4 represents another example of a system for introducing a sample into a chamber. The said system may be used for isolating an analyte from a sample as per one of the embodiments of the present disclosure. This example of the system (1) includes a cartridge comprising a chamber (2) comprising an opening (3) on the second side of the chamber located adjacent to the top end. Adjacent to the chamber (2) is a solution dispensing pouch having a first layer (5) and a second layer (6) opposite to the first layer (5). The first layer (5) has a lower tensile strength than the second layer (6). In some embodiments, the chamber (2) and the first layer (5) of the solution dispensing pouch optionally have an adhesive layer (7) between them. An actuator (8) comprising a blunt member (9) is positioned adjacent to the solution dispensing pouch such that the first layer (5) of the solution dispensing pouch (4) is immediately adjacent to the opening (3) on the second side of the chamber (2) and the second layer (6) is adjacent the actuator (8). In other words, the solution dispensing pouch is positioned between the opening (3) on the second side of the chamber (2) and the actuator (8). The actuator (8) is aligned with the opening (3) on the second side of the chamber (2). The blunt member (9) of the actuator (8) applies pressure on the second layer (6) of the pouch (4) such that the second layer (6) is pushed into the first layer (5) causing the first layer (5) to stretch and rupture and the second layer (6) is not ruptured. The first layer (5) is ruptured at a point immediately adjacent to the opening (3).

[0096] In certain aspects, each said chamber in the cartridge comprises two protrusions extending towards the pouch and flanking the opening on the second side and configured to contact the first layer of the pouch and to accommodate the blunt member between the two protrusions when the actuator applies pressure to the pouch.

[0097] FIG. 5 represents one of the embodiments of the first system of present disclosure. This example of the system includes a cartridge comprising a chamber (2) comprising two protrusions (10) positioned adjacent to the opening (3) on the second side of the chamber (2). The two protrusions (10) accommodate the blunt member (9) when the actuator (8) applies pressure. The solution dispensing pouch is positioned between the opening (3) and the actuator (8). The blunt member (9) of the actuator (8) contacts the second layer of the pouch and pushes it into first layer, wherein the first layer then comes in contact with the two protrusions (10) near the opening (3) causing the first layer to stretch and rupture at a point immediately adjacent to the opening (3).

[0098] In some embodiments, the pouch comprises a rigid member positioned or positionable adjacent to the opening such that the blunt member contacts second layer and applies pressure to the first layer via the rigid member.

[0099] In some embodiments, the rigid member is a magnetically responsive solid member. In some embodiments, the rigid member is a paramagnetic bead or ferromagnetic bead.

[0100] Another aspect of the present disclosure includes a second system for introducing a sample into a chamber. The said system may be used for isolating an analyte from a sample. The system comprising: the cartridge; and an actuator comprising a hollow member, wherein each said chamber comprises two protrusions extending towards the pouch and flanking the opening on the second side; the first layer of the pouch is positioned immediately adjacent the opening on the second side and the second layer of the pouch is positioned adjacent the actuator, the hollow member configured for applying pressure on the second layer of the pouch such that the first layer is stretched against the protrusions and ruptures, and the hollow member is aligned with the opening on the second side such that the first layer is ruptured at a point immediately adjacent to the opening.

[0101] FIG. 6 represents one of the embodiments of the second system of present disclosure. This example of the system includes a cartridge comprising a chamber (2) comprising two protrusions (10) positioned adjacent to the opening (3) on the second side. The actuator (8) comprises a hollow member (11) aligned with the opening (3). The solution dispensing pouch is positioned between the opening (3) and the actuator (8). The hollow member (11) of the actuator (8) applies pressure on the second layer of the pouch and pushes it into first layer, wherein the first layer then comes in contact with the two protrusions (10) near the opening (3) causing the first layer to stretch against the protrusions and rupture at a point immediately adjacent to the opening (3).

[0102] In some embodiments, the opening on the second side is positioned at the top end of the chamber or the inlet of the chamber.

[0103] In some embodiments, the first layer of the pouch is a thin aluminum foil, and the second layer is a thick aluminum foil.

[0104] In some embodiments, wherein upon rupture of the first layer, the solution from the pouch enters the chamber via the opening due to gravity.

[0105] In some embodiments, wherein upon rupture of the first layer, the solution from the pouch enters the chamber via the opening due to negative pressure in the chamber.

[0106] In some embodiments, the system comprises a squeezing member configured to squeeze the solution out of the pouch and to the opening.

[0107] In some embodiments, the system comprises a first chamber having a first opening, the pouch is a first pouch, and the solution is a first solution and the system further comprises a second chamber comprising a second opening and a second pouch positioned adjacent to the second chamber, and wherein the second pouch is filled with a second solution.

[0108] In some embodiments, the system further comprises a third chamber comprising a third opening area and a third pouch positioned adjacent the third chamber.

[0109] In some embodiments, the actuator is configured to be positionable adjacent to the second pouch and / or the third pouch.

[0110] In some embodiments, the first chamber comprises a sample introduction region.

[0111] METHODS

[0112] Aspects of the present disclosure include methods of using the system comprising: activating the actuator to apply pressure on the pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the solution from the pouch enters the chamber via the opening.

[0113] Another aspect of the present disclosure includes a method for isolating an analyte from a sample.

[0114] In using the cartridge described herein for isolating an analyte from a sample, such as a biological sample, solution dispensing pouches are adhered adjacent to each of the said chambers, that is, in the cartridge of FIG. 2, a first pouch (401) is positioned adjacent to the first chamber (201), a second pouch (402) is positioned adjacent to the second chamber (202) and a third pouch (403) is positioned adjacent to third chamber (203). Each of the said pouches contains liquid reagents. In one embodiment, the first pouch (401) contains a lysis buffer, the second pouch (402) contains a wash buffer, and the third pouch (403) contains an elution buffer. The additional pouch, if present between the second and third pouch, may contain a further wash buffer.

[0115] A plurality of solid phase carrier particles, such as described above, is introduced into the first chamber (201). The solid carrier particles, as described above, are able to pass through the chambers and channel upon application of an external force. In one embodiment, the particles are magnetic particles or magnetic beads, and the external force is a magnetic force. At least a plurality, and preferably all, of the particles comprise a surface affinity reagent, as described above, which is effective to specifically and reversibly bind an analyte.

[0116] A sample is introduced into the first chamber (201) through a sample introduction region (12) as shown in FIG. 2. After addition of the sample and binding particles (which may be added in either order), the actuator is activated to apply pressure on the first pouch such that the second is pushed against the first layer causing the first layer to rupture so that the lysis buffer from the first pouch enters the first chamber. The sample and lysis buffer get mixed in the first chamber. Mixing may be facilitated by moving the magnetic particles within the chamber by the above referenced externally applied magnetic force. Other means of moving the particles via an externally applied force can be used, including air pressure, vacuum, centrifugal force, or electrical fields for charged molecules or particles.

[0117] Following cell lysis and binding of analyte, if present, to the magnetic particles, the particles are transferred, via the externally applied force, from the first chamber (201) into the second chamber (202) via channel (13). Upon entering the second chamber (202), the wash buffer from second pouch (402) is introduced using actuator as described for the first pouch (401). Similarly, the magnetic particles are then transferred, via the externally applied force, from the second chamber (202) into the third chamber (203) via channel (13). Upon entering the third chamber (203), the elution buffer from third pouch (403) is introduced using actuator as described for the first pouch (401). The third chamber (203) is then evacuated into the the detection wells, wherein the isolated analyte when the analyte is present in the sample is detected.

[0118] In certain aspects, the method comprises: providing the system; introducing the sample into the first chamber; activating the actuator to apply pressure on the pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the first solution from the first pouch enters the first chamber via the first opening area; mixing the sample and first solution in the first chamber; wherein the first chamber comprises magnetic beads configured for binding to the analyte; moving the magnetic beads to the second chamber; moving the actuator to a position adjacent the second pouch; activating the actuator to apply pressure on the second pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the second solution from the second pouch enters the second chamber via the second opening; moving the magnetic beads to the third chamber; moving the actuator to a position adjacent the third pouch; activating the actuator to apply pressure on the third pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the third solution from the third pouch enters the third chamber via the third opening; draining the third solution from the third chamber into the detection wells via the channel connecting the third chamber to the detection wells, wherein the third solution comprises the isolated analyte when the analyte is present in the sample; and detecting the isolated analyte in the detection wells.

[0119] In some embodiments, the first solution is a lysis buffer, second solution is wash buffer, and the third solution is an elution buffer.

[0120] EXEMPLARY NON-LIMITING ASPECTS OF THE DISCLOSURE

[0121] Aspects, including embodiments, of the present subject matter described above may be beneficial alone or in combination, with one or more other aspects or embodiments. Without limiting the foregoing description, certain non-limiting aspects of the disclosure are provided below. As will be apparent to those of ordinary skill in the art upon reading this disclosure, each of the individually numbered aspects may be used or combined with any one of the preceding or following individually numbered aspects. This is intended to provide support for all such combinations of aspects and is not limited to combinations of aspects explicitly provided below. It will be apparent to one of ordinary skill in the art that various changes and modifications can be made without departing from the spirit or scope of the invention.

[0122] 1. A cartridge comprising a plurality of chambers, each chamber of the plurality of chambers configured for holding a solution and comprising a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a channel connected to the inlet of each chamber.

[0123] 2. The cartridge of aspect 1, wherein the plurality of chambers comprises a first chamber configured for filling with a first solution; a second chamber configured for filling with a second solution; and a third chamber configured for filling with a third solution.

[0124] 3. The cartridge of aspect 2, wherein the first solution is a lysis buffer; the second solution is a wash buffer; and the third solution is an elution buffer.

[0125] 4. The cartridge of any one of aspects 1-3, wherein the inlet of the first chamber is positioned lower than the inlet of the second and third chambers. 5. The cartridge of any one of aspects 1-3, wherein the inlets of the plurality of chambers are located at substantially the same height with reference to the channel.

[0126] 6. The cartridge of any one of aspects 1-5, wherein the channel is substantially parallel to the top ends of the chambers.

[0127] 7. The cartridge of any one of aspects 1-6, wherein the first chamber has a substantially trapezoidal shape.

[0128] 8. The cartridge of any one of aspects 1 -7, wherein the first chamber comprises paramagnetic particles.

[0129] 9. The cartridge of any one of aspects 1-8, wherein the second chamber has a substantially square shape.

[0130] 10. The cartridge of any one of aspects 1-9, wherein the third chamber has a substantially pentagonal shape.

[0131] 11. The cartridge of any one of aspects 1-10, wherein the third chamber comprises an opening at a bottom end of the chamber, wherein the opening is configured for draining the third chamber.

[0132] 12. The cartridge of any one of aspects 1-11, wherein the third chamber is connected to a plurality of detection wells via a channel.

[0133] 13. The cartridge of any one of aspects 2-12, comprising an additional chamber, wherein the additional chamber is configured for filling with a wash buffer, wherein the additional chamber is positioned between the second chamber and the third chamber and is connected to the channel.

[0134] 14. The cartridge of any one of aspects 1-13, wherein each said chamber comprises an opening on the second side of the chamber, wherein the opening is located adjacent to the top end or in the inlet of the chamber.

[0135] 15. The cartridge of any one of aspects 1-14, further comprising a plurality of solution dispensing pouches, each solution dispensing pouch of the plurality of solution dispensing pouches comprising a first layer and a second layer opposite to the first layer and enclosing a solution, wherein each of the chambers is associated with a solution dispensing pouch and the first layer of a solution dispensing pouch is positioned immediately adjacent to the second side of a chamber and wherein the first layer ruptures more easily as compared to the second layer.

[0136] 16. The cartridge of aspect 15 , wherein the first layer of each solution dispensing pouch has a lower elongation at break that is lower than the second layer, optionally wherein elongation at break of the first layer is at least 0.5 times lower than the elongation at break of the second layer. 17. The cartridge of aspect 15 or 16, wherein the first layer of the pouch is adhered to the second side of the chamber.

[0137] 18. The cartridge of any one of aspects 15-17, wherein the first layer of the pouch is a thin aluminum foil, and the second layer is a thick aluminum foil, or the first layer of the pouch is a composite sheet of aluminum and a thermoplastic layer and the second layer is a composite sheet of aluminum, the thermoplastic layer and a polymer layer.

[0138] 19. A system, comprising: the cartridge of any one of aspects 15-18; and an actuator, wherein the first layer of pouch is positioned immediately adjacent the opening on the second side of the chamber and the second layer of the pouch is positioned adjacent the actuator, the actuator comprises a blunt member configured for applying pressure on the second layer of the pouch such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured, and the blunt member is aligned with the opening of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening.

[0139] 20. The system of aspect 19, wherein each said chamber in the cartridge comprises two protrusions extending towards the pouch associated with the chamber and flanking the opening on the second side and configured to contact the first layer of the pouch and to accommodate the blunt member between the two protrusions when the actuator applies pressure to the pouch.

[0140] 21. The system of aspect 19, wherein the pouch comprises a rigid member positioned or positionable adjacent to the opening such that the blunt member contacts second layer and applies pressure to the first layer via the rigid member.

[0141] 22. The system of aspect 19, wherein the rigid member is a magnetically responsive solid member.

[0142] 23. The system of aspect 22, wherein the rigid member is a paramagnetic bead or ferromagnetic bead.

[0143] 24. A system, comprising: the cartridge of any one of aspects 15-18; and an actuator comprising a hollow member, wherein each said chamber comprises two protrusions extending towards the pouch associated with the chamber and flanking the opening on the second side; the first layer of the pouch is positioned immediately adjacent the opening on the second side and the second layer of the pouch is positioned adjacent the actuator, the hollow member is configured for applying pressure on the second layer of the pouch such that the first layer is stretched against the protrusions and ruptures, and the hollow member is aligned with the opening on the second side such that the first layer is ruptured at a point immediately adjacent to the opening.

[0144] 25. The system of any one of aspects 19-24, wherein upon rupture of the first layer, the solution from the pouch enters the chamber, with which the pouch is associated, via the opening due to gravity.

[0145] 26. The system of any one of aspects 19-25, wherein upon rupture of the first layer, the solution from the pouch enters the chamber via the opening due to negative pressure in the chamber.

[0146] 27. The system of any one of aspects 19-26, wherein the system comprises a squeezing member configured to squeeze the solution out of the pouch and towards the opening.

[0147] 28. The system of any one of aspects 19-27, wherein the system comprises a first chamber having a first opening, the pouch is a first pouch, and the solution is a first solution and the system further comprises a second chamber comprising a second opening and a second pouch positioned adjacent to the second chamber, and wherein the second pouch is filled with a second solution.

[0148] 29. The system of aspect 28, wherein the system further comprises a third chamber comprising a third opening and a third pouch positioned adjacent the third chamber.

[0149] 30. The system of aspects 28 or 29, wherein the actuator is configured to be positionable adjacent to the pouches.

[0150] 31. The system of any one of aspects 19-29 or the cartridge of any one of aspects 2- 18, wherein the first chamber comprises a sample introduction region, optionally wherein the sample introduction region is located at the top end of the first chamber.

[0151] 32. A method of using the system of any one of aspects 19-31 comprising: activating the actuator to apply pressure on the pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the solution from the pouch enters the chamber via the opening.

[0152] 33. A method for isolating an analyte from a sample, the method comprising: providing a system comprising a cartridge and an actuator, the cartridge comprising: a first chamber and a first pouch associated with the first chamber, the first pouch comprising a first solution, a second chamber and a second pouch associated with the second chamber, the second pouch comprising a second solution, and a third chamber and a third pouch associated with the third chamber, the third pouch comprising a third solution, wherein each chamber is configured for holding a solution and comprises a first side, a second side opposite to the first side, a bottom end opposite to a top end, an inlet at the top end and an opening in the second side, each pouch comprises a first layer and a second layer opposite to the first layer and enclosing a solution, wherein the first layer of a solution dispensing pouch is positioned immediately adjacent to the second side of a chamber; and a channel connected to the inlet of each chamber; the actuator comprising a blunt member configured for applying pressure on the second layer of the pouch such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured, and the blunt member is aligned with the opening of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening; introducing the sample into the first chamber; activating the actuator to apply pressure on the first pouch associated with the first chamber such that the second layer is pushed against the first layer causing the first layer to rupture so that the first solution from the first pouch enters the first chamber via the opening in the second side ; mixing the sample and first solution in the first chamber; wherein the first chamber comprises magnetic beads configured for binding to the analyte; moving the magnetic beads to the second chamber via the channel; moving the actuator to a position adjacent the second pouch; activating the actuator to apply pressure on the second pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the second solution from the second pouch enters the second chamber via the opening in the second side; moving the magnetic beads to the third chamber via the channel; moving the actuator to a position adjacent the third pouch; activating the actuator to apply pressure on the third pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the third solution from the third pouch enters the third chamber via the opening in the second side; and draining the third solution from the third chamber into the detection wells via a channel connecting the third chamber to detection wells, wherein the third solution comprises the isolated analyte when the analyte is present in the sample.

[0153] 34. The method of aspect 33, wherein the method further comprises detecting the isolated analyte in the detection wells, wherein the first solution is a lysis buffer configured for lysing cells or virus, second solution is wash buffer, and the third solution is an elution buffer. While the present invention has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present invention. All such modifications are intended to be within the scope of the claims appended hereto.

Claims

CLAIMSWhat is claimed:

1. A cartridge comprising a plurality of chambers, each chamber of the plurality of chambers configured for holding a solution and comprising a first side, a second side opposite to the first side, a bottom end opposite to a top end, and an inlet at the top end; and a channel connected to the inlet of each chamber.

2. The cartridge of claim 1, wherein the plurality of chambers comprises a first chamber configured for filling with a first solution; a second chamber configured for filling with a second solution; and a third chamber configured for filling with a third solution.

3. The cartridge of claim 2, wherein the first solution is a lysis buffer; the second solution is a wash buffer; and the third solution is an elution buffer.

4. The cartridge of any one of claims 1-3, wherein the inlet of the first chamber is positioned lower than the inlet of the second and third chambers.

5. The cartridge of any one of claims 1-3, wherein the inlets of the plurality of chambers are located at substantially the same height with reference to the channel.

6. The cartridge of any one of claims 1-5, wherein the channel is substantially parallel to the top ends of the chambers.

7. The cartridge of any one of claims 1-6, wherein the first chamber has a substantially trapezoidal shape.

8. The cartridge of any one of claims 1-7, wherein the first chamber comprises paramagnetic particles.

9. The cartridge of any one of claims 1-8, wherein the second chamber has a substantially square shape.

10. The cartridge of any one of claims 1-9, wherein the third chamber has a substantially pentagonal shape.

11. The cartridge of any one of claims 1-10, wherein the third chamber comprises an opening at a bottom end of the chamber, wherein the opening is configured for draining the third chamber.

12. The cartridge of any one of claims 1-11, wherein the third chamber is connected to a plurality of detection wells via a channel.

13. The cartridge of any one of claims 2-12, comprising an additional chamber, wherein the additional chamber is configured for filling with a wash buffer, wherein the additional chamber is positioned between the second chamber and the third chamber and is connected to the channel.

14. The cartridge of any one of claims 1-13, wherein each said chamber comprises an opening on the second side of the chamber, wherein the opening is located adjacent to the top end or in the inlet of the chamber.

15. The cartridge of any one of claims 1-14, further comprising a plurality of solution dispensing pouches, each solution dispensing pouch of the plurality of solution dispensing pouches comprising a first layer and a second layer opposite to the first layer and enclosing a solution, wherein each of the chambers is associated with a solution dispensing pouch and the first layer of a solution dispensing pouch is positioned immediately adjacent to the second side of a chamber and wherein the first layer ruptures more easily as compared to the second layer.

16. The cartridge of claim 15, wherein the first layer of each solution dispensing pouch has a lower elongation at break that is lower than the second layer, optionally wherein elongation at break of the first layer is at least 0.5 times lower than the elongation at break of the second layer.

17. The cartridge of claim 15 or 16, wherein the first layer of the pouch is adhered to the second side of the chamber.

18. The cartridge of any one of claims 15-17, wherein the first layer of the pouch is a thin aluminum foil, and the second layer is a thick aluminum foil, orthe first layer of the pouch is a composite sheet of aluminum and a thermoplastic layer and the second layer is a composite sheet of aluminum, the thermoplastic layer and a polymer layer.

19. A system, comprising: the cartridge of any one of claims 15-18; and an actuator, wherein the first layer of pouch is positioned immediately adjacent the opening on the second side of the chamber and the second layer of the pouch is positioned adjacent the actuator, the actuator comprises a blunt member configured for applying pressure on the second layer of the pouch such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured, and the blunt member is aligned with the opening of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening.

20. The system of claim 19, wherein each said chamber in the cartridge comprises two protrusions extending towards the pouch associated with the chamber and flanking the opening on the second side and configured to contact the first layer of the pouch and to accommodate the blunt member between the two protrusions when the actuator applies pressure to the pouch.

21. The system of claim 19, wherein the pouch comprises a rigid member positioned or positionable adjacent to the opening such that the blunt member contacts second layer and applies pressure to the first layer via the rigid member.

22. The system of claim 19, wherein the rigid member is a magnetically responsive solid member.

23. The system of claim 22, wherein the rigid member is a paramagnetic bead or ferromagnetic bead.

24. A system, comprising: the cartridge of any one of claims 15-18; and an actuator comprising a hollow member,wherein each said chamber comprises two protrusions extending towards the pouch associated with the chamber and flanking the opening on the second side; the first layer of the pouch is positioned immediately adjacent the opening on the second side and the second layer of the pouch is positioned adjacent the actuator, the hollow member is configured for applying pressure on the second layer of the pouch such that the first layer is stretched against the protrusions and ruptures, and the hollow member is aligned with the opening on the second side such that the first layer is ruptured at a point immediately adjacent to the opening.

25. The system of any one of claims 19-24, wherein upon rupture of the first layer, the solution from the pouch enters the chamber, with which the pouch is associated, via the opening due to gravity.

26. The system of any one of claims 19-25, wherein upon rupture of the first layer, the solution from the pouch enters the chamber via the opening due to negative pressure in the chamber.

27. The system of any one of claims 19-26, wherein the system comprises a squeezing member configured to squeeze the solution out of the pouch and towards the opening.

28. The system of any one of claims 19-27, wherein the system comprises a first chamber having a first opening, the pouch is a first pouch, and the solution is a first solution and the system further comprises a second chamber comprising a second opening and a second pouch positioned adjacent to the second chamber, and wherein the second pouch is filled with a second solution.

29. The system of claim 28, wherein the system further comprises a third chamber comprising a third opening and a third pouch positioned adjacent the third chamber.

30. The system of claims 28 or 29, wherein the actuator is configured to be positionable adjacent to the pouches.

31. The system of any one of claims 19-29 or the cartridge of any one of claims 2-18, wherein the first chamber comprises a sample introduction region, optionally wherein the sample introduction region is located at the top end of the first chamber.

32. A method of using the system of any one of claims 19-31 comprising: activating the actuator to apply pressure on the pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the solution from the pouch enters the chamber via the opening.

33. A method for isolating an analyte from a sample, the method comprising: providing a system comprising a cartridge and an actuator, the cartridge comprising: a first chamber and a first pouch associated with the first chamber, the first pouch comprising a first solution, a second chamber and a second pouch associated with the second chamber, the second pouch comprising a second solution, and a third chamber and a third pouch associated with the third chamber, the third pouch comprising a third solution, wherein each chamber is configured for holding a solution and comprises a first side, a second side opposite to the first side, a bottom end opposite to a top end, an inlet at the top end and an opening in the second side, each pouch comprises a first layer and a second layer opposite to the first layer and enclosing a solution, wherein the first layer of a solution dispensing pouch is positioned immediately adjacent to the second side of a chamber; and a channel connected to the inlet of each chamber; the actuator comprising a blunt member configured for applying pressure on the second layer of the pouch such that the second layer is pushed into the first layer causing the first layer to stretch and rupture and wherein the second layer is not ruptured, and the blunt member is aligned with the opening of the chamber such that the first layer is ruptured at a position immediately adjacent to the opening; introducing the sample into the first chamber; activating the actuator to apply pressure on the first pouch associated with the first chamber such that the second layer is pushed against the first layer causing the first layer to rupture so that the first solution from the first pouch enters the first chamber via the opening in the second side ; mixing the sample and first solution in the first chamber; wherein the first chamber comprises magnetic beads configured for binding to the analyte; moving the magnetic beads to the second chamber via the channel;moving the actuator to a position adjacent the second pouch; activating the actuator to apply pressure on the second pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the second solution from the second pouch enters the second chamber via the opening in the second side; moving the magnetic beads to the third chamber via the channel; moving the actuator to a position adjacent the third pouch; activating the actuator to apply pressure on the third pouch such that the second layer is pushed against the first layer causing the first layer to rupture so that the third solution from the third pouch enters the third chamber via the opening in the second side; and draining the third solution from the third chamber into the detection wells via a channel connecting the third chamber to detection wells, wherein the third solution comprises the isolated analyte when the analyte is present in the sample.

34. The method of claim 33, wherein the method further comprises detecting the isolated analyte in the detection wells, wherein the first solution is a lysis buffer configured for lysing cells or virus, second solution is wash buffer, and the third solution is an elution buffer.

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