Patent Application
20190025163
The present disclosure relates generally to the field of medical sample processing, preparation and analysis and more particularly to processing and assaying samples in in vitro medical diagnostic devices suitable for use at a point-of-need (also commonly referred to point-of-care).
There is a recognized and compelling need for the rapid and accurate diagnosis of common infectious diseases in environments that may be remote from a laboratory. While the field of in vitro diagnostics for the testing for infectious pathogens in human patient, animal, and environmental is well established, it is largely confined to centralized laboratory testing in Clinical Laboratory Improvement Amendment (CLIA) rated medium-complexity or high-complexity facilities. Commonplace techniques used in such centralized laboratories include traditional culturing of specimens, immunological assaying using Enzyme-Linked Immunosuppressant Assay (ELISA), nucleic acid testing (such as polymerase chain reaction, PCR), and other methods. These techniques generally all require complex, expensive, and fixed-site instrumentation along with highly skilled laboratorian staff to operate said equipment and are therefore unsuitable for PON applications. The present invention resolves the obstacles to realizing PON medical diagnostic devices suitable for use in decentralized (e.g. out-patient clinic, doctor's office, remote field setting, and other low resource settings) locations.
In accordance with an illustrative embodiment, a sample processing device includes a receiving chamber having a chamber inlet and a chamber outlet. The sample processing device has a metering portion having a metering reservoir fluidly coupled to the chamber outlet, and at least one vessel. The vessel(s) are fluidly coupled to the metering reservoir by a syphon that is operable to actuate upon a preselected volume of liquid being received at the metering reservoir. The syphon includes a syphon inlet and a syphon outlet, the syphon outlet being fluidly coupled to the vessel. The preselected volume of liquid corresponds to a volume of liquid necessary to deliver a metered volume of fluid to the vessel.
In accordance with another illustrative embodiment, a sample processing device reader comprising a first sample processing device holder. The first sample processing device holder has a plurality of viewing areas and a plurality of cylindrical ports for receiving a plurality of vessels of a sample processing device. The sample processing device reader also includes a receiving portion operable to receive and retain a computing device.
In accordance with another illustrative embodiment, a method of processing a sample includes placing a sample in a receiving chamber of a sample processing device. The receiving chamber has a chamber inlet and a chamber outlet. The method includes suspending at least a portion of the sample in a liquid and motivating the liquid from the receiving chamber to a metering reservoir fluidly coupled to the chamber outlet. The method also includes motivating the liquid to a vessel fluidly coupled to the metering reservoir via a syphon, the syphon being operable to actuate upon a preselected volume of liquid being received by the metering reservoir. The syphon includes a syphon inlet and a syphon outlet, the syphon outlet being fluidly coupled to the vessel, and the preselected volume of liquid corresponds to a volume of liquid necessary to deliver a metered volume of fluid to the vessel.
Other features and advantages of the present invention will be apparent from the accompanying drawings and from the detailed description that follows below.
The conventional model for infectious disease diagnosis relies heavily on centralized laboratory testing (e.g. culture, immunoassay, or polymerase chain reaction), which can often take two to four days to provide a reliable result.
The present disclosure relates to a sample processing device, system, and method that are operable to analyze and record data from solid specimens, including but not limited to tissue biopsy, bone fragment, vectors, insects and other organisms and plants in a remote or low resource setting in which traditional laboratory equipment may not be available, or in a more traditional outpatient setting. Such analytics typically involve some sort of sample preparation since the material targeted for analysis may be contained inside of the sample. For example, if the sample is an organism, such as a mosquito, the target material may be inside of the sample's digestive tract or circulatory system. For illustrative purposes, the present disclosure shows a sample as a mosquito, but it is noted that almost any sample may be processed using the disclosed devices, systems, and methods. For example, the sample could be any kind of insect, a tick, worm, leech, plant matter, tissue, bone, or other environmental matter.
The illustrative embodiments include a process that is tailored to minimize resources and to be lightweight. The system and method are highly mobile and field deployable to allow for use in many forward locations, such as a densely forested area, open land or deserts. The devices may be operated without external power (other than, for example, an operator pushing a button). Independent of the field-deployable aspects of the disclosed embodiments, the present disclosure also provides systems and methods for simultaneously performing a molecular and immunological assay on the same collected specimen. A significant benefit of the illustrative systems and methods is that they provide for the ability to analyze proteins and genetic materials included in a specimen in parallel.
As described in more detail below, in some embodiment, a sample processing device includes a cartridge body having an opening in which (for example) a captured mosquito or (in a traditional doctor's office) a tissue biopsy may be placed. To analyze the sample, a lid is closed about the opening and an associated process is implemented that includes macerating the mosquito to gain access to its digestive and circulatory parts and fluids.
In addition to the foregoing, the present disclosure relates to multiplexed point-of-care diagnostic device capable of simultaneously performing molecular and immunological assays from any specimen type in a completely low-complexity (e.g. CLIA waiver) format. In some illustrative embodiments, a specimen processing does not use external power other than that already available from a smartphone and an optional small battery (e.g., a 9V battery, which may be used to power high-efficiency light emitting diodes (LEDs). The exemplary embodiments operate with intuitive and minimal user interaction with a goal of producing a laboratory quality result in a short amount of time (e.g., less than thirty minutes).
The illustrative embodiments work with a one or more sample preparation cartridges or sample processing devices that are operable to facilitate the analysis of biological fluids or solid tissue samples. The representative cartridges and devices and associated systems are operable to perform sample extraction, mixing or reagent metering, support isothermal amplification and support bead-based immunoassays.
In some embodiments, a docking unit, or mating adapter, is used to provide moderate heating via exothermic films, while battery-powered LEDs provide blue excitation light for fluorescence detection. The representative system may incorporate a commercial, off-the-shelf smartphone or similar computing device as an analytical instrument, thereby lessening the need for specialized instrumentation. The representative system is operable to perform simultaneous ELISA and LAMP multiplexed assays in a low-cost, low-complexity point-of-care device enabled by existing, off-the-shelf computing devices. The representative systems may be used to detect infectious diseases in situations where simultaneous detection of the infectious pathogen and assessment of host response markers are needed to inform clinical decisions.
Referring now to
The sample processing device reader 102 may also include a plurality of second sample processing device holders 110, each of which may be configured to receive and hold a sample processing device 106. In some embodiments, the first sample processing device holder 108 or second sample processing device holder may be equipped with a heat source to incubate, denature, anneal or otherwise heat a sample stored within a sample processing device.
Illustrative embodiments of a sample processing device 200 (analogous to the sample processing device 106 of
In some embodiments, the sample processing device 200 includes a cap 204 having a protrusion that forms a piston 238 relative to the receiving chamber 206 when the cap 204 is joined to the body 202. A plate member 240 is disposed within the receiving chamber 206 between the cap 204 and the chamber outlet 210. In some embodiments, the plate may include a boss feature at its base that is received by a bore at the base of the receiving chamber 206 to orient and position the plate member 240 within the receiving chamber 206. A biasing member 242, which may be a coil spring, an air spring, a hydraulic spring, a leaf spring, a torsional spring, a 3-D printed spring, or an injection molded spring, may be positioned at the base of the plate member 240 to urge the plate away from the chamber outlet 210.
In some embodiments, a base of the piston forms a first macerating surface 244 and a second, upward facing surface 246 of the plate forms a second macerating surface. In such embodiments, the first macerating surface 244 and second macerating surface 246 may operate to grind against one another to macerate or pulverize a sample 264 for subsequent processing and/or analysis. In some embodiments, rotation of the cap 204 may result in the grinding motion being applied to the specimen when the specimen is positioned between the first macerating surface and the second macerating surface, as shown in
In some embodiments, the plate 240 comprises an anti-rotation feature (e.g., a key, protrusion or slot) that engages with a corresponding anti-rotation feature of the receiving chamber 206 to restrict rotation of the plate member 240 relative to the receiving chamber 206 while allowing axial movement of the plate member 240 relative to the receiving chamber 206. For example, an anti-rotation feature of the plate member 240 may be tabs that slide axially (relative to the bore of the receiving chamber 206) along grooves formed within the outer boundary of the receiving chamber 206 or a bore at the base of the receiving chamber 206.
The cap 204 of the sample processing device 200 may include a storage reservoir 248 having a storage reservoir inlet 254 and a storage reservoir outlet 252. A temporary seal 250, which may be a frangible member, check valve, or other suitable feature is positioned at the storage reservoir outlet 252. The storage reservoir inlet 254 of the cap 204 is configured to receive a cap piston 256. The cap piston 256 resembles a button and is operable to apply a compressive force to a volume of the storage reservoir 248 to displace a liquid 234 disposed therein.
Upon actuation, the cap piston 256 applies a compressive force to the liquid 234 sufficient to generate a critical pressure differential across the temporary seal 250. The temporary seal 250 is operable to rupture or breach to allow the liquid 234 to pass from the storage reservoir 248 to storage reservoir outlet 252. The storage reservoir outlet 252 is fluidly coupled to the receiving chamber 206 so that liquid 234 forced from the storage reservoir 248 is able to flow into the receiving chamber 206 when the cap piston 256 is depressed. The liquid 234 may be a stabilized carrier or an agent. For example, the liquid 234 may include a buffer, chemical or biological reagent, or a pre-treatment solution such as a lysing agent, a mucolytic agent, a detergent, a surfactant or a host of other liquids.
By generating a hydraulic force, the cap piston 256 is operable to induce fluid flow across the plate member 240 and sample 264. Hence, the cap piston 256 is operable to be depressed by an operator to push liquid 234 through the storage reservoir outlet 252. The liquid 234 may be forced out in a way that impinges on the plate member 240 with sufficient hydraulic or dynamic force to overcome the biasing member 242, thereby inducing separation between the plate member 240 and the base of the piston 238. The moving liquid 234 may thereby be operable to loosen macerated pieces of the sample 264, which are no longer captured between the plates, by moving at an appropriate velocity, to wash macerated pieces of the sample 264 downstream for further processing. In some embodiments, a filter may be in or adjacent to the chamber outlet 210 to prevent particles of the sample 264 that are larger than a predetermined size from entering the metering portion 212 (discussed below).
In some embodiments, one or both of the first surface 244 of the cap piston 256 and the second surface 246 of the plate member 240 may include channels to facilitate fluid flow. In some embodiments, the first surface 244 and second surface 246 may be textured or include an alternative feature (e.g., a roller, blade, perforation, grate, squeegee, other compressive tool) to grind the sample 264 or facilitate extraction of ground or pulverized specimen from the receiving chamber 206. In some embodiments, the first surface 244 and second surface 246 may have, for example, textured surfaces having a grit number of (for example) between 60 and 220, or any other suitable range. In other embodiments, the first surface 244 and second surface 246 may have interlocking, radial grooves that grate against one another when the cap 204 is rotated relative to the body 202.
In some embodiments, the cap 204 may have an internal cam 262 that is sized and configured to engage an external cam 260 of the receiving chamber inlet 208 so that, as the cap 204 is placed over the chamber inlet 208 and rotated, the internal cam 262 will engage the external cam 260 to draw the cap 204 toward the body 202 of the specimen processing device 200. In some embodiments, complementary magnetic components may be installed on or integrated within the cap 204 and body 202 to attract the cap 204 toward the body.
Referring now to
In some embodiments, the vessels may be considered assaying vessels that are used to perform an immunological assay or a molecular assay. As referenced herein, an immunological assay is an assay in which a protein associated with the immune system such as an antibody, is used as the biomarker (or target analyte) for a particular disease state (e.g., the presence of a virus or other disease). Correspondingly, a molecular assay is an assay in which a form of genetic material, such as an oligonucleotide is used as the biomarker for a particular disease state. To facilitate execution of a molecular assay, one or more of the assaying vessels may be prepopulated with a reagent solution, which may include, for example, a lyophilized reagent that is operable to facilitate the execution of a loop-mediated isothermal amplification assay. To facilitate execution of an immunological assay, one or more of the other assaying vessels may be prepopulated with fluorescently tagged beads. Such beads may be magnetic, such as, for example, super-paramagnetic microspheres.
For the case of a molecular assay, each designated vessel will contain lyophilized, loop-mediated isothermal amplification-oligonucleotide strand displacement (LAMP-OSD) probes pre-populated as part of a manufacturing step. Upon delivery of a metered amount of reagent from the syphon and weir system (metering portion) described herein, the lyophilized LAMP-OSD probes will be reconstituted, thereby enabling the progression of LAMP biochemistry.
For the case of an immunological assay, each designated vessel will contain lyophilized, super-paramagnetic microspheres (which may alternatively be referred to as “beads”) which have been functionalized with an appropriate capture moiety as part of a manufacturing step. Upon delivery of a metered amount of reagent from the syphon and weir system, the lyophilized super-paramagnetic microspheres become reconstituted, thereby enabling the progression of ELISA biochemistry. In such progression, a magnet is used to pull beads to a particular surface of a vessel such that the beads are in the focal plane of a photosensor (e.g., a CMOS sensor). In this manner, background from the supernatant will not deleteriously impact the signal-to-noise ratio of the subsequent spectroscopic measurement. Blue LEDs may be used to excite (or pump) a fluorophore that has been previously functionalized on the bead to excite a fluorescence response which may be measured by the photosensor.
As referenced herein, a syphon is understood to be a tube, pipe, hose, or other conduit having a shorter leg at a higher level and a longer leg extending to a lower level through which a liquid can be moved from the higher level to the lower level by atmospheric pressure forcing the liquid up the shorter leg while the weight of the fluid in the longer leg causes continuous downward flow. Referring again to the figures, the syphon 222 is configured to actuate and dispense fluid to a vessel 270, 271, 272 once a predetermined volume of liquid is received at the metering reservoir. To that end, the syphon 222 includes a syphon cap 228 overlying a conduit to form a syphon inlet 224. The conduit terminates in a syphon outlet 226, which is fluidly coupled to, and operable to deliver a liquid to the vessel (e.g., vessel 270). In some embodiments, the predetermined volume of liquid corresponds to a volume of liquid necessary to deliver a metered volume of fluid to each of a plurality of vessels 270, 271, 272.
The metering reservoir 220 may include an interchange weir 230, the height of which (in connection with the functional area of the metering reservoir) determines the volume of liquid needed to initiate the syphon 222. In some embodiments, the metering reservoir includes an overflow weir 232. The overflow weir 232 may be positioned to allow liquid to run off from the metering reservoir 220 if fluid in excess of the predetermined volume is received at the metering reservoir 220.
In some embodiments, the sample processing device includes a plurality of vessels 270, 271, 272, each coupled to the metering reservoir 220 by a plurality of syphons 222. The plurality of syphons 222 may be formed by a plurality of conduits covered by one or more syphon caps 228 to form a fluid flow path that generates a syphon. The syphon may be configured to deliver a metered and equivalent volume of liquid to each of the plurality of vessels 270, 271, 272. The metered and equivalent volume may correspond to the height of the interchange weir, the diameter of the syphon and the surface properties of the materials that make up the syphon (i.e., depending on whether such materials are hydrophobic or hydrophilic.
Referring again to the sample processing device reader 102 of
In some embodiments, the second sample processing device holder(s) 110 may include a heating element, which may be a resistive or similar heating element. The heating element may be controlled by, for example, a manually controlled thermostat, and may be powered by a local battery or comparable power source.
An illustrative sample holder 401 that includes a heating element is described with regard to
To facilitate completion of a molecular assay, the sample holder 401 may include a permanent magnet 421 and an electromagnet 423 located proximate to apertures 402 receiving a plurality of assaying vessels associated with a molecular assay. In addition, the sample holder 401 may include an actuator 425, controller and power source to facilitate operation of the electromagnet 423. The actuator 425 may be manually operated or may be operated remotely from an electronic device (e.g., computing device 104) that is being used to complete the molecular or immunological assay.
An alternative embodiment of a sample processing device 500 that includes an integrated specimen processing cartridge is shown in
In
The sample processing device 500 is shown as including an actuator 514 which may form a portion of a collection fluid reservoir 520, such that the actuator 514 may be depressed by a user to initiate a fluid flow process. To that end, the actuator 514 is operable to move fluid a sample collection liquid within the sample processing device 500 when the housing 505 is in the closed state and the actuator 514 is depressed or otherwise actuated. In the illustrated embodiment, the actuator 514 is a bulb-type actuator. In other embodiments, a piston-cylinder, electronic actuator, or other suitable type of actuator may be used.
The sample collection liquid may be stored within the collection liquid reservoir 520 prior to actuation and held in place by a destructible seal that prevents the sample collection liquid from exiting the first reservoir 520 through a port that is fluidly coupled to the specimen receiving chamber 534 while the seal is intact. Actuation of the actuator 514 may result in destruction (e.g., by pressurization or otherwise) of the destructible seal to allow for the communication of sample collection liquid from the collection liquid 520 to the specimen receiving chamber 534. A locking mechanism to hold the actuator 514 in the engaged position, or a check valve may be included at the referenced port so that once the actuator 514 has been engaged and sample collection liquid evacuated from the first reservoir 520, the sample collection liquid may not return to the first reservoir 520.
In some embodiments, the first reservoir 520 may be a blister pack. In others, the first reservoir 520 may be a cylinder, a liquid-filled bulb, or other similar container that may be evacuated when a hydraulic force is applied upon actuation by the actuator 514. The first reservoir 520 may be pre-filled with a sample collection liquid, which is a fluid that is selected to strip sample from a specimen collection device and carry the sample in suspended, diluted, or dissolved form through the fluid flow paths described below. The sample collection liquid may include a reagent such as a lysing agent to react with the sample, an elution buffer, an anti-coagulant or a solvent. The actuator 514 is operable to generate a hydraulic force to propel the sample preparation fluid from the first reservoir, through a tube, and through the specimen receiving chamber 534, and ultimately to a downstream reservoir, in the form of a metering portion 512 that distributes the sample preparation fluid into vessels 570-574 for subsequent processing and analysis (See
The specimen receiving chamber 534 may be sized and configured to receive a specimen collection device (e.g., a swab) and facilitate extraction of the sample from the specimen collection device. In some embodiments, the specimen receiving chamber 534 includes features that roil the sample collection liquid passing through the specimen receiving chamber 534. As referenced herein, “roiling” refers to the manipulation of a fluid in a manner that induces turbulence or increased fluid shear forces to facilitate extraction of a specimen for analysis from a specimen collection device. For example, the specimen receiving chamber 534 may include helical or spiral features that induce vortex flow or spiral-like flow patterns, such as grooves. Such features may induce high fluid shear forces in the sample preparation fluid, increase turbulent energy and result in a higher Reynolds number. The foregoing characteristics may be understood to enhance the ability to extract a specimen for analysis from a specimen collection device placed in the chamber. In some embodiments, the roiling features may be spiralized grooves that engage or nearly engage the shaft of the specimen collection device to force the sample preparation fluid to follow a helical flow path through the sample collection portion of the specimen collection device as it is propelled through the specimen receiving chamber 534. For clarity, it is noted that in the context the sample preparation described in this disclosure, “extraction” does not relate to pulling DNA from a cell. Instead, “extraction” refers generally to the ability to recover organisms, molecules or other particles of interest off from a collection device and deliver those particles to a subsequent stage for further analysis.
The shaft portion of a swab or other suitable sample collection device may be deemed a nuisance once the sample is acquired and placed within the sample processing device 500. To facilitate removal of the portion of the shaft that extends beyond the housing 505 of the sample processing device 500, a swab may have a pre-scored shaft to facilitate breaking off the protruding portion of the swab shaft. Alternatively, the sample processing device 500 may include a swab cutter 512 to neatly trim away excess swab shaft material. Moreover, a swab shaft seal may be included where the shaft is inserted into the housing 505 of the sample processing device 500 and the adjoining external boundaries of specimen receiving chamber 534 may include a similar gasket or other suitable seal to form a sealed liquid flow path through the specimen receiving chamber 534.
A representative system 610 that includes a sample processing device 600 is shown in
The forgoing system and sample processing device may be operated to process and analyze a sample in accordance with any number of illustrative methods. Referring again to
In some embodiments, the method includes attaching a cap 204 to a body 202 of the specimen processing device 200, wherein the cap 204 has a protrusion that forms a piston 238 relative to the receiving chamber 206. The method further includes applying a compressive force to the sample 264 between the piston 238 and a plate member 240 disposed within the receiving chamber 206 between the cap 204 and the chamber outlet 210.
In some embodiments, a first surface 244 of the piston 238 forms a first macerating surface, and wherein a second surface 246 of the plate member 240 forms a second macerating surface. In such embodiments, the method may further include rotating the cap 204 relative to the plate member 240 to apply a compressive force upon the sample 264 and grind the sample 264 between the first surface 244 and the second surface 246.
The method may also include suspending at least a portion of the sample 264 in a liquid 234. This suspending step may include injecting the liquid 234 from a storage reservoir 248 and across the receiving chamber 206 containing the sample 264. To effect suspension of the sample 264, the cap 204 includes a storage reservoir 248 having a liquid 234 disposed therein. The liquid is contained within the storage reservoir 248 by a temporary seal 250 and a cap piston 256. The cap piston 256 is operable to compress a volume of the storage reservoir 248 to pressurize and displace the liquid 234 from the storage reservoir 248. Displacing the liquid 234 may be accomplished by depressing the cap piston 256 to displace the liquid 234, thereby generating a sufficient pressure differential to breach the temporary seal 250 and force the liquid 234 to flow through the storage reservoir outlet 252 and across the first surface 244 and second surface 246 to extract and suspend at least a portion of the sample 264.
The illustrative method also includes motivating the liquid 234 from the receiving chamber 206 to a metering reservoir 220 fluidly coupled to the chamber outlet 210. In some embodiments, the method includes using one or more syphons to distribute a preselected volume of the liquid 234 (which now includes suspended bits of the sample 264) into one or more vessels 270, 271, 272. As noted above, the syphon is operable to actuate upon a preselected volume of liquid being received by metering reservoir 220.
Where the sample processing device 200 includes a plurality of vessels 270, 271, 272, the syphon 222 is operable to distribute an approximately equivalent amount of liquid 234 to each vessel 270, 271, 272.
In some embodiments, the method further includes disposing the sample processing device 200 in a sample processing device reader 102 (see
In an embodiment in which the sample processing device reader 102 includes a receiving portion 122, an illustrative method may also include placing a computing device 104 in the receiving portion 122 of the sample processing device reader 102 by securing the computing device 104 with a clamp 120. Here, the method may also include aligning a camera, CMOS sensor, CCD, FPA, MCP/PMT, or other similar photo-detecting device of the computing device 104 with a focal region of the first sample processing device holder 108.
In an embodiment in which the sample processing device reader 102 includes one or more second sample processing device holders 110 having heating elements, the method may also include heating vessels of one or more second sample processing devices 110 to incubate the samples disposed therein and to prepare them for analysis. Similar to the second sample processing device holders 110, the first sample processing device holders 108 may also include a heating element that may be used to incubate a sample for analysis or to facilitate the analysis process in other ways.
In some embodiments, the plurality of viewing areas 112 include one or more illumination source(s), and the associated method includes illuminating each of the plurality of viewing areas. In some embodiments, the method includes using the computing device 104 to obtain a measurement of the sample at each of the plurality of viewing areas 112, 114, 116.
Referring again to
Next, the user places the sample collection device 700 into one of three active heating positions on the MIDA docking unit 712 and starts a customized mobile application. At this point, the elution-including solution flows into the metering portion described above to provide metered amounts of the sample extract into each reaction tube or vessel pre-loaded with assay reagents. The target-specific nucleic acids are then amplified by loop-mediated isothermal amplification (LAMP) and transduced into fluorescence via oligonucleotide strand displacement (OSD) probes, while protein-based biomarkers are captured using magnetic immunoassay beads. The user then transfers the sample processing device into the imaging position (see right-most sample processing device 100), where fluorescence excitation is achieved using inexpensive light-emitting diodes and the associated emission is acquired with a smartphone's CMOS sensor, both of which are aligned and optically focused by the docking unit/mating adapter. Ultimately, the mobile application will provide prompts to the user when the test is complete while custom signal processing algorithms will analyze the data and output results.
Target-specific nucleic acids may be amplified in illustrative systems using a variant of LAMP that has evolved from the field of DNA computation, with the design of oligonucleotide strand displacement (OSD) probes. These OSD probes are hemi-duplex DNA that undergo sequence-specific toehold-mediated strand exchange with LAMP amplicons resulting in the separation of a fluorophore and quencher. The OSD probes yield the same level of surety for LAMP as do TaqMan probes for PCR, but without increasing the operational complexity of the assay thereby enabling viral and/or cell lysis concurrently with amplification upon heating at 60° C., eliminating the need for separate nucleic acid purification from clinical samples. Furthermore, the OSD probes can be excited by low-cost LEDs that will be located in the sample processing system and the resulting fluorescence, filtered through >500 nm bandpass gel filters, may be detected by an off-the-shelf smartphone. By coupling the signal amplification power of LAMP-OSD to the imaging, data processing, and porting power of smartphones, our technology will be ideally positioned to deliver sophisticated molecular diagnostics in a user-friendly, inexpensive, and universally accessible format.
The sample processing system may then be operated to carry out magnetic bead-based assay using a sandwich immunoassay format with an immobilized target-specific capture antibody and fluorescently-tagged detecting antibody. In any given vessel, two different analytes can be multiplexed by using two sets of magnetic beads and matched pair antibodies conjugated to distinct fluorophores, such as Cyber Green and Phycoerythrin (PE). Both of these reporter molecules can be efficiently excited by deep blue LED's. The imaging portion (i.e., the portion of the vessel that is monitored by the visual sensor of the computing device) of the sample processing system will also contain magnetic elements directly adjacent to the relevant vessels, thereby separating and concentrating analyte-bound fluorescent probe/magnetic bead complexes into a spatially-resolved location away from the depleted remainder of unbound detecting antibodies.
Changes in the fluorescent intensity profile across the width of each optically transparent vessel may be used to determine quantitative concentration of the given analyte. Various reagent vessels geometries may be used. For example, the vessels may have angled, flat walls will be explored to further improve the magnetic separation and imaging at the ideal optical focal plane to enhance signal-to-background ratios. It is important to recognize that we are not attempting to image individual beads, but rather an aggregated intensity value of all bead-bound fluorescence. It is this intensity value which will be used, through the prior generation of standards curves, to give a quantitative measurement of analyte concentration.
A siphon and weir system (metering portion) is used in each of the sample processing device embodiments described above and operates to apportion a collected sample eluate in three-five equal volumes, without the use of electrical energy or any user intervention. Equal distribution of the sample helps to ensure that assays are quantitative via interpolation of fluorescence intensity values against a known standard curve. As discussed previously, the metering functions by depositing sample eluate into a bulk sample reservoir. The sample liquid then flows from this reservoir into the dispensing chambers in unequal volumes but will level out across a series of weirs until it reaches an overflow weir that is located at a higher elevation. As the level in each dispensing chamber rises, the sample mixture flows into the syphon cap. Once the liquid level within the syphon cap is above the drain tube, the sample begins to flow through the drain tube, forming a “water lock”, which causes sample to steadily drain downstream. When the outer liquid level is below the syphon cap, air is sucked into the syphon cap, causing the water lock to break and flow into the drain tube to stop immediately, thus separating the sample into equally divided portions and depositing these portions into each of the reaction and detection vessels.
It is noted that unless an embodiment is expressly stated as being incompatible with other embodiments, the concepts and features described with respect to each embodiment may be applicable to and applied in connection with concepts and features described in the other embodiments without departing from the scope of this disclosure. To that end, the above-disclosed embodiments have been presented for purposes of illustration and to enable one of ordinary skill in the art to practice the disclosure, but the disclosure is not intended to be exhaustive or limited to the forms disclosed. Many insubstantial modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the disclosure. For example, it should be understood that the above-mentioned metering portion, which is operable to distribute metered amounts of a liquid, may function separate and distinctly from the receiving chamber and cap. To that end, the metering portion may be coupled to any suitable upstream processing configuration that dispenses a fluid for metering, and the cap and receiving chamber may be coupled to any suitable downstream processing configuration that collects or receives a liquid that has been processed in accordance with the foregoing description. The scope of the claims is intended to broadly cover the disclosed embodiments and any such modification, including without limitation the following examples:
A sample processing device comprising: a receiving chamber having a chamber inlet and a chamber outlet; a metering portion having a metering reservoir fluidly coupled to the chamber outlet; and a vessel fluidly coupled to the metering reservoir by a syphon, the syphon being operable to actuate upon a preselected volume of liquid being received at the metering reservoir, wherein the syphon comprises a syphon inlet and a syphon outlet, the syphon outlet being fluidly coupled to the vessel, and wherein the preselected volume of liquid corresponds to a volume of liquid necessary to deliver a metered volume of fluid to the vessel.
The sample processing device of example 1, wherein the metering reservoir comprises an interchange weir.
The sample processing device of example 1, wherein the metering reservoir comprises an overflow weir.
The sample processing device of example 1, wherein the syphon inlet comprises a syphon cap.
The sample processing device of example 1, wherein: the metering reservoir comprises an interchange weir, the vessel comprises a plurality of vessels coupled to the metering reservoir by a plurality of syphons, each of the plurality of vessels being fluidly coupled to a syphon, wherein the preselected volume of liquid corresponds to a volume of liquid necessary to deliver a metered and equivalent volume of liquid to each of the plurality of vessels, and wherein the metered and equivalent volume corresponds to the height of the interchange weir (and the diameter of the syphon and the surface properties of the syphon).
The sample processing device of example 1, further comprising: a cap having a protrusion that forms a piston relative to the receiving chamber when the cap is joined to the receiving chamber; a plate disposed within the receiving chamber between the cap and the chamber outlet; and a biasing member operable to urge the plate away from the chamber outlet.
The sample processing device of example 6, wherein the biasing member is selected from the group consisting of a coil spring, an air spring, a hydraulic spring, a leaf spring, a torsional spring, a 3-D printed spring, and an injection molded spring.
The sample processing device of example 6, wherein a base of the piston forms a first macerating surface, and wherein a second surface of the plate forms a second macerating surface.
The sample processing device of example 8, wherein rotation of the cap results in a grinding motion being applied to a sample positioned between the first macerating surface and the second macerating surface.
The sample processing device of example 8, wherein the plate comprises an anti-rotation feature that engages with a corresponding anti-rotation feature of the receiving chamber to restrict rotation of the plate relative to the chamber while allowing axial movement of the plate relative to the chamber.
The sample processing device of example 10, wherein the anti-rotation feature of the plate comprises tabs and wherein the corresponding anti-rotation feature of the receiving chamber comprises grooves.
The sample processing device of example 6, wherein the cap comprises a storage reservoir, a temporary seal (e.g., a frangible seal or a check valve) positioned at a storage reservoir outlet, the cap further comprising an inlet for receiving a cap piston, the cap piston being operable to compress a volume of the storage reservoir to displace a liquid disposed within the storage reservoir.
The sample processing device of example 12, wherein the cap piston is operable to displace the liquid, thereby breaching the temporary seal and causing the liquid to flow through the storage reservoir outlet, and across the first macerating surface and second macerating surface to extract at least a portion of a sample positioned therebetween, and through the chamber outlet.
The sample processing device of example 13, wherein the cap piston is operable to generate fluid flow that induces separation of the plate relative to the piston to allow more sample to become suspended in the liquid.
The sample processing device of example 1, further comprising a filter positioned between the chamber outlet and the metering portion.
The sample processing device of example 15, wherein at least one of the first surface of the cap piston and the second surface of the plate comprises channels.
The sample processing device of example 6, wherein the cap comprises an internal cam that is sized and configured to engage an external cam of the receiving chamber such that rotation of the cap relative to the receiving chamber results in the cap being forced toward the receiving chamber.
The sample processing device of example 6, wherein the cap comprises a magnetic engagement feature that is sized and configured to attract and engage a complementary magnetic engagement feature of the receiving chamber.
The sample processing device of example 8, wherein at least one of the first macerating surface and second macerating surface is textured or includes an alternative grinding feature such as a roller; grinder; squeegee, or other compressive tool.
The sample processing device of example 20, wherein the first macerating surface and second macerating surface have a grit number of between 60 and 220.
The sample processing device of example 20, wherein the first macerating surface and second macerating surface comprise interlocking, radial grooves.
The sample processing device of example 20, wherein the second macerating surface is perforated.
The sample processing device of example 12, wherein the liquid disposed within the storage reservoir comprises a preselected volume of liquid selected from the group consisting of a stabilized carrier or an agent.
The sample processing device of example 1, wherein the syphon comprises a hydrophobic coating.
The sample processing device of example 1, wherein the syphon comprises a hydrophilic coating.
A sample processing device reader comprising: a first sample processing device holder having one or more viewing areas and one or more cylindrical ports for receiving one or more vessels of a sample processing device; and a receiving portion operable to receive and retain a computing device (or measurement device).
The sample processing device reader of example 27, wherein the receiving portion comprises a plurality of rails and a clamping mechanism, the clamping mechanism being operable to substantially align a photo detecting device of the computing device with a focal region of the first sample processing device holder.
The sample processing device reader of example 27, further comprising one or more second sample processing device holders having one or more second cylindrical ports for receiving one or more second vessels of one or more second sample processing devices.
The sample processing device reader of example 29, wherein each of the one or more second sample processing device holders comprises a heating element.
The sample processing device reader of example 27, wherein each of the plurality of viewing areas comprises an illumination source.
The sample processing device reader of example 31, wherein each of the illumination sources is a LED.
A system for processing a sample, the system comprising a sample processing device reader in accordance with any of examples 27-32 and a sample processing device in accordance with any one of examples 1-26.
A method of processing a sample, the method comprising: placing a sample in a receiving chamber of a sample processing device having a chamber inlet and a chamber outlet; suspending at least a portion of the sample in a liquid; motivating the liquid from the receiving chamber to a metering reservoir fluidly coupled to the chamber outlet; and motivating the liquid to a vessel fluidly coupled to the metering reservoir via a syphon, the syphon being operable to actuate upon a preselected volume of liquid being received by the metering reservoir, wherein the syphon comprises a syphon inlet and a syphon outlet, the syphon outlet being fluidly coupled to the vessel, and wherein the preselected volume of liquid corresponds to a volume of liquid necessary to deliver a metered volume of fluid to the vessel.
The method of example 34, wherein the vessel comprises a plurality of vessels coupled to the receiving chamber by a plurality of syphons, each vessel being fluidly coupled to a syphon, the method further comprising motivating the liquid to the vessel comprises motivating a metered and equivalent volume of liquid to each of the plurality of vessels when a preselected volume of fluid is received in the metering reservoir, the preselected volume corresponding at least in part to the position of an interchange weir positioned within the metering reservoir.
The method of example 34, wherein the sample processing device further comprises a cap having a protrusion that forms a piston relative to the receiving chamber when the cap is joined to the receiving chamber and a plate disposed within the receiving chamber between the cap and the chamber outlet, the method further comprising applying a compressive force to the sample between the piston and the plate.
The method of example 36, wherein a base of the piston forms a first macerating surface, and wherein a second surface of the plate forms a second macerating surface, and wherein the method further comprises rotating the cap relative to the receiving chamber to grind the sample between the first macerating surface and the second macerating surface.
The method of example 36, wherein the cap comprises a storage reservoir, a temporary seal (a frangible seal or a check valve) positioned at a storage reservoir outlet, the cap further comprising an inlet for receiving a cap piston, the cap piston being operable to compress a volume of the storage reservoir to displace a liquid disposed within the storage reservoir, wherein the method further comprises depressing the cap piston to displace the liquid, thereby breaching the temporary seal and causing the liquid to flow through the storage reservoir outlet, and across the first macerating surface and second macerating surface to extract at least a portion of a sample positioned therebetween (to cause the sample to be suspended in the liquid), and through the chamber outlet.
The method of example 34, further comprising disposing the sample processing device in a sample processing device reader having a first sample processing device holder comprising a plurality of viewing areas and a plurality of cylindrical ports for receiving a plurality of vessels of the sample processing device, the sample processing device reader further comprising a receiving portion operable to receive and retain a computing device (or measurement device).
The method of example 39, further comprising placing a computing device in the receiving portion of the sample processing device reader by securing the computing device with a clamping mechanism of the sample processing device reader.
The method of example 40, further comprising aligning a photo detecting device of the computing device with a focal region of the first sample processing device holder.
The method of example 41, wherein the sample processing device reader further includes one or more second sample processing device holders having one or more second cylindrical ports for receiving one or more second vessels of one or more second sample processing devices, wherein each of the one or more second sample processing device holders comprises a heating element, the method further comprising heating the plurality of second vessels of one or more second sample processing devices.
The method of example 34, wherein each of the plurality of viewing areas comprises an illumination source, the method further comprising illuminating each of the plurality of viewing areas.
The method of example 43, further comprising using the computing device to obtain a measurement of the sample at each of the plurality of viewing areas.
The sample processing device of example 1, wherein the syphon comprises a hydrophilic coating and a hydrophobic coating.
As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprise” and/or “comprising,” when used in this specification and/or the claims, specify the presence of stated features, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and/or groups thereof. In addition, the steps and components described in the above embodiments and figures are merely illustrative and do not imply that any particular step or component is a requirement of a claimed embodiment.
This application is a continuation-in-part of U.S. patent application Ser. No. 15/664,781, which is a continuation of U.S. patent application Ser. No. 15/419,816, now U.S. Pat. No. 9,719,892, entitled PROCESSING DEVICE FOR PROCESSING A HIGHLY VISCOUS SAMPLE and filed on Jan. 30, 2017, which are incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| Parent | 15419816 | Jan 2017 | US |
| Child | 15664781 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 15664781 | Jul 2017 | US |
| Child | 16130736 | US |
