INTEGRATED, POINT OF SALE, BLOOD TESTING SYSTEMS AND METHODS

Abstract

An integrated system for automatically analyzing in real time an analyte in a sample containing fluid is disclosed. The system includes a fluid separator for receiving the sample and separating therefrom a fluid component that contains the analyte, a non-optical, chemical analyte sensing device having at least one sensor for chemically analyzing the analyte, and a microfluidic channel for transferring at least a portion of the fluid component from the separator to the sensing device. In a preferred embodiment, the system is point of care, single-use cartridge in a base unit that separates plasma from a small sample of whole blood and tests an analyte of interest in the plasma.

Claims

1. An integrated, automated system for analyzing in real time an analyte in the plasma of a sample of whole blood, comprising: a. a blood separator for receiving the whole blood sample and separating blood plasma therefrom; b. a microfluidic channel fluidly connected to the separator for transmitting at least a portion of the plasma from the separator; and c. a non-optical, chemical analyte sensing device that receives and analyzes plasma from the microfluidic channel.

2. The system of claim 1, wherein the whole blood sample comprises less than 1 milliliter of whole blood.

3. The system of claim 1, wherein the sample comprises between 20 microliters and 1 milliliter of whole blood.

4. The system of claim 1, wherein the microfluidic channel actively transmits the portion of the plasma.

5. The system of claim 1, wherein the microfluidic channel passively transmits the portion of plasma.

6. The automated system of claim 1, wherein the chemical analyte sensing device is a biosensor microchip that generates an electrical signal from a bio-chemical reaction in the plasma.

7. An integrated system for analyzing in real time an analyte in a sample containing fluid, comprising: a. a fluid separator for receiving the sample and separating therefrom a fluid component that contains the analyte; b. a non-optical, chemical analyte sensing device having at least one sensor for chemically analyzing the analyte; and c. a microfluidic channel fluidly connecting the separator to the non-optical, chemical analyte sensing device for transferring at least a portion of the fluid component from the separator to the sensing device.

8. An apparatus for testing in real time an analyte in a sample of whole blood, comprising: a. a blood separator for receiving the sample and separating therefrom blood plasma; b. a biosensor microchip having at least one biosensor for detecting and analyzing, label-free, at least one analyte in the plasma; and c. a microfluidic subsystem fluidly connecting the separator and the microchip, the subsystem having a channel for transmitting a portion of the plasma from the separator onto the biosensor.

9. The apparatus of claim 8, wherein the biosensor comprises a nanowire field effect transistor (nwFET).

10. The apparatus of claim 8, wherein the microchip contains a plurality of biosensors.

11. The apparatus of claim 10, wherein the microchip is multiplexed such that each biosensor is adapted to detect a different analyte in the plasma.

12. The apparatus of claim 10, wherein the microchip is adapted to detect multiple analytes in the plasma simultaneously.

13. The apparatus of claim 8, wherein the separator, microchip and microfluidic subsystem are contained in a cartridge.

14. The apparatus of claim 8, wherein the cartridge is a point-of-care (POC), single-use and disposable cartridge.

15. The apparatus of claim 8, further including a reagent processing well connected to the microfluidic subsystem.

16. The apparatus of claim 8 wherein the blood separator is a centrifuge.

17. The apparatus of claim 13, further includes an electronic base unit connected to the cartridge.

18. The apparatus of claim 17, wherein the cartridge is removably connected to the electronic base unit.

19. The apparatus of claim 17, wherein the base unit comprises a. a power source; b. a circuit board electronically connected to the microchip for receiving electronic signals from the microchip representative of the detected analyte as results data; c. a display screen for displaying results of the testing; and d. a control unit for controlling the results data from the circuit board and the display screen.

20. The apparatus of claim 18, further including an electric motor removably connected to the blood separator in the cartridge for driving the separator.

21. The apparatus of claim 18, wherein the electric motor is connected to a pressure subsystem that creates in the microfluidic subsystem negative or positive pressure to compel movement of plasma across the microfluidic channel and toward the microchip.

22. The apparatus of claim 19, further including a wireless module to transfer results data external to the base unit.

23. The apparatus of claim 19, wherein the control unit electronically controls the sequence and timing of movement of fluid from the separator area to the microchip.

24. The apparatus of claim 19, wherein the control unit electronically controls the timing and movement of reagents through the microfluidic system.

25. The apparatus of claim 17, wherein the base unit is connected to a second cartridge as claimed in claim 6, the base unit capable of processing and reading samples obtained from both cartridges containing different blood samples.

26. A single use cartridge for conducting a plurality of immunologic and DNA/RNA/protein testing assays in real-time on charged molecules, ions, and/or chemicals in a whole blood sample obtained from a subject, the cartridge comprising: a. a whole blood sample processing device for processing the sample; b. a semiconductor, label-free assay microprocessor detection chip containing biosensor detection wells capable of detecting analytes, oligos or other molecules in the processed blood sample; c. a receiving cavity configured to receive the blood sample and provide the sample to the processing device; and d. a microfluidic system that transmits at least a portion of the processed sample from the processing apparatus to the microprocessor detection wells.

27. The cartridge of claim 26, wherein the sample comprises less than 1 milliliter of whole blood.

28. The cartridge of claim 26, wherein the sample comprises between 20 microliters and 1 milliliter of whole blood.

29. A method for processing in real time whole blood in a self-contained cartridge, the method comprising: a. depositing a sample of whole blood into a blood separator in the cartridge; b. separating the sample in constituent parts to isolate plasma in the sample; c. drawing, via microfluid transmission, a portion of the plasma toward a bio-sensing microchip in the cartridge; d. detecting an analyte in plasma deposited on a biosensor disposed on the microchip; and e. transmitting an electrical signal representative of the detected analyte to a processor to be recorded and/or displayed as digital data.

30. A system for testing a sample for an analyte of interest, comprising: a. a separator for receiving the sample and separating therefrom a fluid for testing; b. a microchip having at least one sensor for chemically detecting, label-free, at least one analyte in the fluid; and c. a microfluidic subsystem fluidly connecting the separator and the microchip, the subsystem having a channel for transmitting a portion of the fluid from the separator on a sensor on the microchip.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Further advantages of the present invention may become apparent to those skilled in the art with the benefit of the following detailed description of the preferred embodiments and upon reference to the accompanying drawings in which:

[0032] FIG. 1 is a diagrammatic side view of one embodiment of a fluid separation and analysis system in a cartridge according to the present invention:

[0033] FIG. 2 is a diagrammatic top view of one embodiment of a microfluidic “lab on a chip” system (MLOC) used by at least one embodiment of the present invention;

[0034] FIG. 3 is a diagrammatic side view of one specific embodiment of the system in a cartridge of FIG. 1, wherein the fluid separator comprises a centrifugation system.

[0035] FIG. 4 is diagrammatic side view of one embodiment of the system of the present invention comprising the cartridge shown in FIG. 3 connected to a base unit;

[0036] FIG. 5 is a top view of the system shown in FIG. 4; and

[0037] FIG. 6 is flow diagram showing one method of the present invention as implemented with the system of FIGS. 4 and 5.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0038] Referring now to the drawings, like reference numerals designate identical or corresponding features throughout the several views. The present invention discloses an integrated system for analyzing in real time an analyte in a sample containing liquid. It should be understood that the present invention can be implemented for analyzing analytes in biologic fluids (such as blood or urine) or non-biological fluids that require a fluid separation stage as a precursor to chemically analyzing the separated fluid of interest.

[0039] The system comprises a fluid separator 210 for receiving the sample and separating therefrom a fluid component that contains the analyte; a non-optical, chemical analyte sensing device 50 having at least one sensor for chemically analyzing the analyte; and a microfluidic channel 180 fluidly connecting the separator to the non-optical, chemical analyte sensing device for transferring at least a portion of the fluid component from the separator to the sensing device. Combining and interconnecting these processes—i.e. fluid separation, transmission via the one or more microfluidic channels to the analyte sensing device, and analysis by the sensing device—into one integrated package is what enable the sample to be analyzed in real time. As used throughout, “real time” means the actual time during which all of these processes in the integrated system occurs. This is to be understood in contrast with conventional blood processing that does not occur in not real time, where the places and times of blood collection, separation and plasma analysis may all be different. steps of blood separation In practice, “real time” could be mere minutes or even seconds.

[0040] FIG. 1 shows diagrammatically a side view of the components of one embodiment of an all-in-one, automated separator/analyzer of the present invention in the form of a cartridge. As seen, the cartridge 10, has a sample inlet port 70 connected to a liquid separator 210 for directly loading (inserting or injecting) therein a relatively small amount of the sample. The cartridge includes a chemical analyte sensing device 50 that is fluidly connected to an output valve 160 via the microfluidic channel system 180. In the preferred embodiment of the system, the components are integrated together in a single-use, disposable self-contained cartridge, having first processed the sample in separator 210 and then analyzed the separated fluid on a single use chemical analyte sensing device 50.

[0041] The invention will now be described as implemented for one preferred system embodiment, namely a system for processing a patient's whole blood for the analysis of multiple components in the patient's blood plasma. In such a system, the present invention discloses an integrated, automated system 10 for analyzing in real time an analyte in the plasma of a sample of whole blood. This system comprises a blood separator 210 for receiving the whole blood sample and separating blood plasma therefrom; a microfluidic channel 180 fluidly connected to the separator for transmitting at least a portion of the plasma from the separator; and a non-optical, chemical plasma analyte sensing device 50 that receives and analyzes plasma from the microfluidic channel. The whole blood sample may comprise less than 1 milliliter of whole blood and preferably between 20 microliters and 1 milliliter of whole blood. The microfluidic channel may actively or passively transmit the portion of the plasma to the sensing device.

[0042] A relatively new technological advance that holds great promise to revolutionize the field of biological fluid or “biofluidics” analysis and diagnostics, and that enables one embodiment of the present invention, is the development of label-free, semiconductor biosensor microchips integrated with microfluidics devices for the analysis of fluid samples. These small form factor labs-on-a-chip (LOC) can offer low cost, fast, label-free, highly sensitive yet not fragile, sensing and chemical analysis of analytes in small samples of fluids such as blood plasma. Using LOC's enable small and portable form factors, such as lightweight tabletop systems and even battery powered systems. These new biosensor microchips comprise multiple highly sensitive biosensor transistors—such as those disclosed in U.S. Pat. No. 9,645,135, titled “Nanowire field-effect transistor biosensor with improved sensitivity”—designed on a very small semiconductor chip, or microchip. These new generation of sensors can now (a) directly detect with good sensitivity and scalability and quantify any number of biological molecules (analytes) deposited on their surfaces; (b) be multiplexed,—meaning multiple biosensors can reside on a single chip, with each sensor capable of being prepared with a different reagent to test for a different chemical constituent, all done simultaneously, and (c) convert these results into electrical signals (data) for further processing and readout.

[0043] The inventor of the present invention has recognized that such a biosensing LOC device can be designed in a microfluidic system as the non-optical, chemical analyte sensing device of the present invention. Thus, coupling such a device to a small form factor fluid separation system 210, such as a mini-centrifugal or membrane-type blood plasma extraction system, via one or more microfluidic channels 180, all packaged in an integrated unit, like a sterile cartridge, creates a portable, low cost, disposable, truly point-of-care, plasma-separating and analyte detection system that can truly revolutionize and disrupt the entire blood testing industry. FIG. 2 shows one exemplary implementation of a non-optical, chemical analyte sensing device 50 shown in FIG. 1, namely, a microfluidic biosensing “lab on a chip” (MLOC) device 50. As diagrammatically shown, fluid sample containing analytes of interest (biologic or otherwise) is drawn into a microfluidic system 180 at inlet 60 and through microfluidic channels 170, 175 of microfluidic system 180. This fluid transport channels are overlaid on a multiplexed lab on a biosensing microchip LOC 50, such as one of the biosensing chips designed by Selfa, Inc., a portfolio company of the California NanoSystems Institute (CNSI). This flow causes small drops of the sample flowing through the channels to be deposited on sensing zones, or “wells,” 190 of highly sensitive, label-free, multiwire nanowire field effect transistor (mwFET's) biosensors disposed on the chip surface, with each well 190 being prepared with a reactant specific for a measurement of interest. Each well 190 can thus be prepared to analyze a different analyte, simultaneously (i.e., multiplexed). These wells chemically react with the biomolecules that are deposited thereon in order to analyze them for the specific analyte being tested for. The reactions in each well in turn generates electrical signals on the microchip indicative of the analyte reaction in that well, hence providing electronically recordable and readable test results. Thus, in the blood plasma testing use case described herein, plasma fluid travels through channels 170 and 175, depositing along the way plasma fluid on all the wells 190, each prepared with a reactant designed for a specific blood test.

[0044] In one preferred embodiment of the non-optical, chemical analyte sensing device 50, a semiconductor LOC chip 50 comprises a surface that contains a biological layer with multiple wells containing antibodies or oligonucleotides or any other molecule used to test specific analytes or other target molecules that may be loaded into the wells.

[0045] The present inventive system thus combines into a single package any suitable non-optical chemical analyte sensing device, such as a LOC described herein, with any suitable blood separation technology that can be fluidly connected to the analyte sensing device and packaged therewith in a relatively compact and preferably disposable package, or cartridge. While the following embodiments show this aspect of the invention in the form of a cartridge, it should be understood however, that the form of device is not essential to the present invention, and persons of ordinary skill in the art can readily select a suitable form for a given application. Thus, while the term “cartridge” will be used hereinafter, it should be understood to mean any such suitable form for this combined microfluidic separator/analyzer package. Further, the microfluidic cartridge of the present invention may be constructed from any suitable material, such as a sterile, transparent plastic, mylar or latex, using any method such as injection molding or lamination, and it may be made as a disposable package for one-time use, or otherwise.

[0046] Turning now to the liquid separator, any known separator technology that effectively in real time separates fluid containing an analyte of interest from a sample may be used. In plasma use case, the whole blood separator built into the cartridge of the present invention may be any of the new test-tubeless blood separation technologies that can in real time and in small form factor separate plasma from whole blood sample. Non-limiting examples include centrifuge technologies, such as the plasma centrifugation technology designed by Sandstone Diagnostics, any plasma separator member device (e.g, from Pall Corporation or from Spot On Sciences, Inc.), microfluidic filter systems that draw whole blood through the filter using any known drawing method (such as with piezoelectric pumps, micro-syringe pumps, electroosmotic pumps, and the like, or those driven by inherently available internal forces as gravity, hydrostatic pressure, capillary force, absorption by porous material or chemically induced pressures or vacuum, including the microfluidic systems described in U.S. Pat. No. 7,419,638 to Micronics, Inc.), or any other plasma separating and collecting device that can be suitably designed with a micro-fluidic technology to supply the plasma to the biosensing LOC.

[0047] Accordingly, FIG. 3 shows a side view of a specific implementation of the disposable cartridge 10 shown in FIG. 1, with the blood separator 210 implemented as a miniaturized centrifuge 80 (and its components 30, 40 and 90) that engages a motor connectable to the cartridge, such as the centrifuge designed by Sandstone Diagnostics. This is explained in further detail in connection with FIGS. 4 and 5.

[0048] FIG. 4 shows a side view and FIG. 5 shows a top view of a real time, analyte diagnostic Point of Care (POC) system 200 of the present invention, comprising the disposable cartridge 10 shown in FIG. 3, as physically placed in and mechanically and electrically connected to an electrically-powered base unit 120. As seen in all three figures, the cartridge 10 has a blood access port 70 connected to the small centrifuge 80 having a centrifuge disc 40 for directly loading (inserting or injecting) therein (with no test tube) a relatively small amount of a patient's whole blood. As seen in FIGS. 3 and 4, four (4) guides 30 hold the disc 40 in place when rotating at high speed. The underside of disc 40 is connected to a fixed, rotatable rod 90, which, when engaged in base unit 20 (FIG. 4), engages motor 100 for translating the rotation of the motor when activated to the centrifuge 80. As described above, cartridge 10 includes a non-optical, chemical analyte sensing device 50 such as biosensor microprocessor chip, LOC 50, that is fluidly connected to plasma output valve 160 via the microfluidic channel system 180. In the preferred embodiment, the cartridge 10 is a single-use, disposable self-contained cartridge, having processed the patient's blood on a single use centrifuge 80 and then analyzed the plasma on the single-use biosensor microprocessor chip 50.

[0049] In one preferred embodiment of the real time, analyte diagnostic Point of Care (POC) system 200, the base unit 120 contains a power source (not shown), a motor 100, an electronic control unit 110, a circuit board 130, a visual display 150 for displays test results, and, preferably, a wireless communications module 140. Alternatively, or additionally, the unit 20 may include storage (not shown) for digitally storing results of testing. It will be understood that base 20 can be powered by any suitable power source (e.g, AC or battery) and its electronics can comprise any conventional electronics components that can be designed and programmed as needed in a small form factor (e.g., portable or table-top) to achieve the desired actions (e.g., programmably driving the motor 100 via unit 110) and the desired results (e.g., designing the circuit board 130 to process the signals from LOC 50, programming the controller 110 to receive the analyte data from board 130 and drive the display 150 to displaying test results).

[0050] Flow diagram 300 in FIG. 6 shows the operation of the POC testing system of the present invention according to the embodiments shown in FIGS. 2-5. In step 302, a small amount of whole blood is loaded into the cartridge 10, and specifically into the blood separator 210 (or 80) via inlet port 70. From this point forward, the process is fully automated and is completely self-contained and thus sterile. Upon powering on the POC system, blood separator, in step 304, engages the sample to automatically separate out the blood cells, leaving the plasma to be processed. In the case of the centrifuge, when the cartridge 10 secured to the base 20, and is loaded with whole blood, the base 20 may be turned on (automatically or manually) and engaged via the electronic control unit 110. The motor 100 then spins the centrifuge 80 rapidly for a prescribed or programmed period of time (e.g. for less than 90 seconds) via rod 90, separating the blood so that the plasma is extractable. In this embodiment, in step 306, the electronic control unit 110 then opens the valve 160 on the microfluidics transfer channel 180, and activates, in step 308 the motor 100 to produce negative pressure through the tubing system 180 that extends over analyte testing device LOC 50. Thus, in step 310 plasma that was drawn through the fluid transfer channel 180 bathes the biosensing wells 190 on the chip 50 (FIG. 2). In step 312, a biochemical reaction occurs on each of the wells 190. This is where the “magic” happens, whereby in a preferred embodiment that uses multiplexed biosensors, the sensors of each well simultaneously test the analytes desired for, and chip 50 convert the results into electric signals that are sent to the circuit board 130 on base unit 20 for processing. In step 314, the circuit board 130 is programmed to collect and compile the signals as results data which is then—driven by controller 110—visually displayed on the screen 150. The data may optionally be stored in storage, and/or sent out in step 316 to remote storage or to directly a physician wireless device or lab via wireless communications module 140.

[0051] While embodiments of the invention have been illustrated and described, it is not intended that these embodiments illustrate and describe all possible forms of the invention. Various changes, modifications, and alterations in the teachings of the present invention may be contemplated by those skilled in the art without departing from the intended spirit and scope thereof. It is intended that the present invention encompass such changes and modifications.