EXTRACTION CONTAINER

Abstract

An extraction container includes a molded tubular body having openings at opposing first and second end thereof. A base cap is configured to obstruct a base opening disposed at the base of the tubular body. The base cap is integrally molded with the tubular body and connected thereto by a frangible connection. The base cap can be removed by breaking the frangible connection to de-obstruct the second opening. The first base cap and tubular body include integrally molded engagement features to permit the base cap to be reattached to the tubular body to re-obstruct the base opening. A removable foil or polymer seal obstructs a top opening disposed at the top end of the tubular body. A top cap and hinge are integrally molded with the tubular body and configured to re-obstruct the top opening following removal of the seal.

Claims

1. A method comprising: removing a removable seal sealing an access opening of a generally tubular body thereby exposing a liquid disposed within the generally tubular container; combining a sample comprising a target with the exposed liquid thereby forming a liquid mixture of the sample and the liquid within the generally tubular body; resealing the access opening with a lid comprising (i) a dispensing opening and (ii) a securing component that obstructs the dispensing opening and is secured to the lid via a frangible connection; with the access opening resealed with the lid, heating the liquid mixture within the generally tubular container to a temperature of at least about 100° C., wherein the lid and the securing component essentially prevent liquid of the liquid mixture or gas within the generally tubular body from leaking or escaping therefrom during the heating; and after heating the liquid mixture, opening the dispensing opening by breaking the frangible connection between the securing component and the lid and then dispensing at least some of the liquid mixture from the generally tubular body through the dispensing opening.

2. The method of claim 1, wherein the generally tubular container and the lid including the securing component are made of a polymer.

3. The method of claim 1, wherein the securing component is a cap and the method further includes resecuring the cap to the lid to reseal the dispensing opening after dispensing the at least some liquid mixture.

4. The method of claim 1, wherein the method further includes subjecting the at least some dispensed liquid mixture to an assay to determine the presence and/or amount of the target.

5. The method of claim 4, wherein the target is indicative of a pathogen.

6. The method of claim 1, wherein the step of heating comprises subjecting the liquid mixture within the generally tubular body to microwave radiation.

7. The method of claim 1, wherein the step of heating comprises heating the liquid mixture to a temperature and for a duration sufficient to lyse bacterial cells within the liquid mixture.

8. The method of claim 7, wherein the step of heating comprises heating the liquid mixture to a temperature and for a duration sufficient to lyse bacterial cells of Mycobacterium tuberculosis and/or a bacterium of the genus Streptococcus.

9. The method of claim 8, wherein the target is indicative of at least one of Mycobacterium tuberculosis and/or a bacterium of the genus Streptococcus and the method further includes subjecting the at least some dispensed liquid mixture to an assay to determine the presence and/or amount of the target.

10. The method of claim 1, wherein the heating step comprises heating the liquid mixture for a period of less than about 200 seconds and more than about 60 seconds.

11. The method of claim 1, wherein the heating step comprises heating the liquid mixture to a temperature that is at least about 115° C. and less than about 150° C.

12. The method of claim 1, wherein the total volume of the liquid mixture within the tubular body during the heating step is less than about 50 .Math.L.

13. The method of claim 1, wherein, during the heating step, no more than about 0.5% of the total mass of the liquid mixture present at the start of the heating step escapes or leaks from the tubular body.

14. The method of claim 13, wherein, during the heating step, less than about 0.25% of the total mass of the liquid mixture present at the start of the heating step escapes or leaks from the tubular body.

15. The method of claim 1, wherein the step of dispensing comprises squeezing an exterior surface of the tubular body between fingers of a user to increase a gas pressure therein and dispense the at least some liquid mixture through the dispensing opening.

16. The method of claim 1, wherein the lid comprises a filter and the step of dispensing comprises passing the at least some liquid mixture through the filter.

17. The method of claim 1, wherein the liquid within the tubular body comprises at least one of buffer, universal transport media (UTM), Viral Transport Media (VTM) or lysis medium.

18. The method of claim 1, wherein the sample comprises at least one of saliva, mucous, sputum, or blood.

19. The method of claim 1, wherein the removable seal comprises one or more layers comprising a liquid-impermeable metal foil and/or a liquid-impermeable polymer film.

20. The method of claim 1, wherein an interior surface of the tubular body defines a multifoil.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0056] FIG. 1 illustrates an embodiment of an extraction container of the invention;

[0057] FIG. 2 illustrates an exploded view of the extraction container of FIG. 1 with a foil seal covering a top opening of a tubular body of the extraction container having been removed from the top opening and a filter having been removed from within the tubular body;

[0058] FIG. 3 illustrates an assembly process for manufacturing the extraction container of FIG. 1;

[0059] FIG. 4 illustrates a process for using the extraction container of FIG. 1;

[0060] FIG. 5 illustrates another embodiment of an extraction container and a method of pooling samples of the invention; and

[0061] FIG. 6 illustrates a method for pooling samples of the invention.

DETAILED DESCRIPTION

[0062] With reference to FIGS. 1 and 2, an extraction container includes a generally tubular body, a base cap disposed at a base of the tubular body, a seal obstructing a top opening at the top of the tubular body, a top cap connected to the tubular body via a hinge, and a filter disposed within the tubular body adjacent the base thereof. A liquid, such as an extraction buffer, is disposed within the tubular body. The generally tubular body, base cap, top cap and hinge are integrally formed, e.g., molded, and composed of a polymer material such as polypropylene, polyethylene, or cycloolefin.

[0063] A proximal portion of the base cap is connected to the tubular body via a frangible connection. When connected to the tubular body via the frangible connection, the base cap obstructs a base opening disposed in the base of the tubular body and prevents the liquid from exiting the tubular body via the base opening. When the base cap is removed from the tubular body by breaking the frangible connection (see, e.g., the discussion below with respect to FIG. 4), liquid within the tubular body may be dispensed therefrom through the base opening. The exterior of the base of the tubular body and an interior of a distal portion of the base cap include corresponding engagement features, e.g., threads, projections and notches, or combination thereof, so that the base cap can be reattached to the tubular body by engaging the engagement features to again obstruct the base opening and prevent further exit of liquid from the tubular body (see, e.g., the discussion below with respect to FIG. 4).

[0064] The filter is typically composed of a polymeric material and configured to permit the passage of liquid and a target to be determined in a diagnostic determination but to obstruct the passage of concomitant materials that might degrade the performance, accuracy, and/or precision of the diagnostic determination. For example, the filter may be configured to obstruct the passage of fibers or other parts of a swab that may inadvertently be released into the extraction liquid within the tubular body. The filter is disposed within the tubular body, e.g., adjacent the base thereof, so that liquid exiting the tubular container through the base opening must first pass through the filter. The filter is disposed within the tubular body without requiring additional components, e.g., by press-fitting the filter in place.

[0065] The seal is typically composed of a metallic foil, a polymer layer, e.g., a polymer film, or combination thereof and is releasably secured to the top of the tubular body to prevent liquid from exiting the tubular body through the top opening. The seal prevents the passage of the liquid through the top opening as well as preventing the evaporation of the liquid therein. The seal may be heat staked to the tubular body. An exemplary seal is a multilayer seal having a thickness of about 0.025 mm and formed of polymer-metal-polymer layers, e.g., polyethylene-aluminum-polyethylene.

[0066] The top cap is secured to the tubular body via a hinge and may be used to re-obstruct the top opening after the seal has been removed to prevent liquid from exiting the tubular body through the top opening.

[0067] With reference to FIG. 3, a method for assembling the extraction container includes (1) integrally forming, e.g., by molding, the tubular body, base cap, top cap, and hinge, (1) pressing the filter into the tubular body, (3) adding an extraction liquid to the tubular body, and (4) sealing the top opening with the seal. In embodiments, the method for assembling the functional components of the extraction container consists essentially of the aforementioned steps. By functional components, it is meant the components of the extraction container in the form in which the extraction container is provided to an end user and which materially contribute to the functioning of the container in use. In a kit including such an extraction container, other components such as a swab, diagnostic cartridge, labels, or instructions would not be considered functional components of the extraction container itself.

[0068] With reference to FIG. 4, a method for using the extraction container includes (1) removing the seal from the top opening thereof, (2) inserting a sample collection device, e.g., a swab, carrying a biological sample into the liquid within the tubular body and manipulating the tubular body as by squeezing the exterior surface between the user’s fingers to extract some of the sample into the liquid to form a mixture, (3) removing the sample collection device and sealing the top opening with the top cap, (4) separating the base cap from the tubular body by breaking the frangible connection as by twisting the base cap, (5) dispensing some of the mixture through the filter and base opening, e.g., onto a sample application port of a diagnostic device, and (6) closing the base cap by engaging the engagement features of the base cap and tubular body. The mixture within the tubular body is then contained to prevent contamination of the user and their surroundings and also permits the mixture to be preserved for later analysis. The method of using the extraction container may further include subjecting the mixture to a heating step prior to the step of breaking the frangible connection of the cap. The heating may be performed using any of the techniques disclosed herein, e.g., by subjecting the container to microwave radiation. The heating may be performed to a temperature and for a duration sufficient to lyse cells of the biological sample and release contents therefrom. For example, the heating temperature and duration may be as disclosed herein. The heating step may be performed without removing the entirety of the sample collection device from the collection device. For example, the collection device may be a swab and the user may break a shaft of the swab thereby leaving a portion within the extraction container prior to sealing the top opening.

[0069] With reference to FIG. 5, an extraction container has a generally tubular body defining a longitudinal axis and including (i) a closed end and an open end spaced apart from one another along the longitudinal axis and (ii) an interior. A buffer liquid, e.g., VTM and/or a lysing medium, occupies a lower portion of the interior of the tubular body. The lower portion of the interior has an inner surface that is spaced apart radially from the longitudinal axis and defines a cinquefoil having five (5) lobes. A seal, e.g., as described for the embodiment of FIGS. 1 and 2, is releasably secured to the top of the open end of the tubular body to prevent liquid from exiting the tubular body through the opening during shipping and storage. The extraction container also includes a lid configured to close the open end. The lid has a passage configured, when in an open state, to permit liquid within the tubular body to pass through the passage and exit the tubular body. A cap is secured to the lid via a frangible connection and disposed to obstruct the passage, e.g., as described for the embodiment of FIGS. 1 and 2. When the cap is removed by breaking the frangible connection, the passage is opened. The cap and lid each include at least one respective engagement feature that can be engaged such that the cap again obstructs the passage, e.g., as shown in FIG. 4.

[0070] Continuing with FIG. 5, a method of using the extraction container to prepare a pooled sample may proceed as follows. A user removes the seal, e.g., by peeling, to expose the interior of the tubular member and the liquid disposed therein. The user provides a first swab that has a swab shaft and a collection swab tip at a distal end of the shaft. A portion of the swab shaft proximal to the collection swab tip is frangible to permit the shaft to be broken, separating the distal portion of the swab shaft from the proximal portion of the swab shaft. The collection swab tip will have been used to obtain a sample from a first subject, e.g., a first human being. The sample may be, e.g., a nasal or a nasopharyngeal sample. The first swab is inserted into the tubular member until at least a portion, substantially all, or essentially all of the collection swab tip is immersed in the liquid. The user then breaks the swab shaft of the first swab at the frangible connection so that the distal portion of the swab shaft including the collection swab tip is retained within the tubular member. A proximal portion of the swab shaft retained within the tubular member may include a portion of the frangible connection at which the shaft was broken.

[0071] The user then provides a second swab, the second swab having a collection swab tip that has been used to obtain a sample from a second, different subject, e.g., a second human being. Using the second swab, the user repeats the steps of inserting the swab into the liquid and breaking the frangible connection to leave the distal portion of the second swab in the liquid along with the distal portion of the first swab. The forgoing steps may repeated, e.g., an additional three times, each using a swab that has been used to obtain a sample from a respective different subject, e.g., a respective different human being. Typically, the samples from the different subjects are of the same type.

[0072] The cinquefoil interior of the extraction container includes five lobes, each of which at least partially accommodates a collection swab tip when inserted therein. Because of the lobed shape, the volume of liquid required to immerse the collection swab tips is less than would be required if the interior were generally circular. Accordingly, the liquid resulting from the pooling of the multiple samples will have a higher concentration of any targets. In addition, the lobes guide the user when inserting successive swabs and provide a visual indicator of when five swabs have been inserted.

[0073] The position of the frangible portion of the swab shafts is chosen so that lid can be closed even with the distal portions of the swabs retained within the tubular body. The user closes the lid and then mixes the liquid, which forms a mixture including five pooled samples. After mixing and optional incubation, the user breaks the cap from the lid, dispenses an amount of sample for analysis, and then uses the cap to once again seal the pooled sample liquid within the extraction container. The method may further include subjecting the mixture with pooled samples to a heating step prior to the step of breaking the frangible connection of the cap. The heating may be performed using any of the techniques disclosed herein, e.g., by subjecting the container to microwave radiation. The heating may be performed to a temperature and for a duration sufficient to lyse cells of the biological sample and release contents therefrom. For example, the heating temperature and duration may be as disclosed herein.

[0074] The cinquefoil is a multifoil that has a number N = five lobes. As an alternative, the extraction container may have an interior with a multifoil that is, e.g., a trefoil (3 lobes), quatrefoil (4 lobes), sexfoil (6 lobes), septfoil (7 lobes), or octofoil (8 lobes). The interior surface of each lobe may be, for example, arcuate (e.g., circular or elliptical) or linear (e.g., wedge”<” shaped). Turning to FIG. 6, a method of using an extraction container to prepare a pooled sample may proceed as follows. The extraction container includes an extraction buffer disposed therein and a seal, as disclosed herein. A user removes the seal, e.g., by peeling, to expose the interior of the tubular member of the extraction container and the liquid disposed therein. The user provides a first swab that has a swab shaft and a collection swab tip at a distal end of the shaft. The collection swab tip will have been used to obtain a sample from a first subject, e.g., a first human being. The sample may be, e.g., a nasal or a nasopharyngeal sample. The first swab is inserted into the tubular member until at least a portion, substantially all, or essentially all of the collection swab tip is immersed in the liquid. The user then agitates or stirs the swab within the liquid for a period of time, e.g., 10 seconds, to combine the sample with the liquid. The user then removes the first swab from the extraction container, squeezing the exterior of the extraction vial, e.g., between the user’s fingers, to compress the collection swab tip during removal to extract liquid absorbed thereon.

[0075] Once the first swab has been removed from the extraction container, the user provides a second swab, the second swab having a collection swab tip that has been used to obtain a sample from a second different subject, e.g., a second human being. Using the second swab, the user repeats the steps of inserting the swab into the liquid and agitating or stirring the swab to combine the sample of the second swab with the liquid and first sample within the extraction container. The forgoing steps may repeated, e.g., an additional three times, each using a swab that has been used to obtain a sample from a respective different subject, e.g., a respective different human being. Typically, the samples from the different subjects are of the same type. The resulting liquid in the extraction container is a mixture including at least some of the sample from each of the swabs. A portion of the mixture may be removed from the extraction container as disclosed herein, e.g., by breaking the frangible connection of the cap to expose a dispensing opening, and subjected to an assay for the determination of one or more targets therein. The method may further include subjecting the mixture to a heating step prior to the step of breaking the frangible connection of the cap. The heating may be performed using any of the techniques disclosed herein, e.g., by subjecting the container to microwave radiation. The heating may be performed to a temperature and for a duration sufficient to lyse cells of the biological sample and release contents therefrom. For example, the heating temperature and duration may be as disclosed herein.

[0076] In embodiments, the functional components of an extraction container as disclosed herein consist essentially of the integrally formed, e.g., molded, tubular body and base cap. In some embodiments, the functional components of the extraction container further consist essentially of a filter disposed within such tubular body with an integrally formed first cap, which may be at the base or top of the extraction container. In still further embodiments, the functional components of the extraction container further consist of the liquid disposed within such tubular body, a seal covering a top opening of the tubular body, and, optionally, a second cap and hinge integrally formed with the tubular body and first cap, where the second cap is configured to reseal the top opening of the tubular body after removing the seal.

[0077] In embodiments, an extraction container consists of the following functional components: an integrally formed, e.g., molded, tubular body with base cap connected to the tubular body via a frangible connection, a filter disposed within the tubular body and configured such that liquid exiting a base opening of the tubular body must pass through the filter, an extraction liquid disposed within the tubular body, and a seal covering a top opening of the tubular body. In embodiments the integrally formed tubular body and base cap further include a top cap connected to the tubular body via a hinge with the top cap and hinge being integrally formed with the tubular body and base cap.

[0078] A tubular or generally tubular body may define a cross section about a longitudinal axis. The cross section may be, e.g., arcuate such as circular or elliptical, and/or include linear features such as a polygonal cross section. A multifoil may include arcuate and/or linear features.

[0079] In embodiments, a kit includes any of the extraction containers as disclosed herein, a sample collection device such as a swab, and a diagnostic cartridge such as a microfluidic device, and methods for using same, as described in U.S. Provisional Application No. 62/991,446 filed Mar. 18, 2020 or U.S. Provisional Application No. 63/067,782 filed Aug. 19, 2020 (the ‘782 application) and International Patent Application No. PCT/US2021/013325 (“the ‘325 Application”) titled “Fluid Control in Microfluidic Devices” and filed Jan. 13, 2021, and/or diagnostic reagents and methods for determination of a pathogen as described in U.S. Provisional Application No. 62/992,681 filed Mar. 20, 2020, International Patent Application No. PCT/US2021/023532, filed Mar. 22, 2021, and/or the ‘325 application. Each of these applications is incorporated herein by reference in its entirety. The extraction container may be used to apply a mixture of extraction buffer and sample to the diagnostic cartridge to permit the determination of a target, e.g., a pathogen or target indicative of the presence of a pathogen, in the sample. In some embodiments, the kit includes an extraction container and a number N collection swabs, where N is at least the number of samples to be pooled using the extraction container. In embodiments in which the extraction container includes a multifoil, the number N of swabs may be the same as the number of the multifoil.

[0080] An immunological assay may be performed using a sample that is a mixture including at least a portion of a sample obtained from each of a plurality of subjects, e.g., mammals. Each of the subjects or mammals may be a human being. A determination of the presence of a pathogen in the mixture is indicative that the pathogen was present in the sample obtained from at least one of the plurality of subjects. A determination of the absence of a pathogen in the mixture is indicative that the pathogen was absent from each of the samples obtained from each of the plurality of subjects. Such a multi-subject assay can be used, for example, to reduce the total number of assays required to surveil the presence of an infectious disease in a population of subjects.

[0081] The immunological assay may be an assay for the determination of a target, e.g., a biomolecule such as an antigen, antibody, or protein, indicative of the presence of an infectious disease. For example, the assay may be any of the assays disclosed or claimed in the ‘325 application, e.g., as in any of claims 124-139 or 176-214 thereof, in which the “sample” is a mixture comprising samples from each of the plurality of subjects. The assay may be performed using any of the microfluidic devices and systems disclosed or claimed in the ‘325 application, e.g., in any of claims 140-156.

[0082] The number of subjects “N” may be, for example, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, or at least about 10. The number “N” may be, for example, about 25 or less, about 20 or less, about 15 or less, about 10 or less, about 9, or less, about 8, or less, about 7 or less, about 6 or less, or about 5 or less.

INCORPORATION BY REFERENCE

[0083] The entire disclosure of each of the patent and scientific documents referred to herein is incorporated by reference for all purposes.

EQUIVALENTS

[0084] The invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting on the invention described herein. Scope of the invention is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.