JP-2026076207-A - Automated point-of-care device for complex sample processing and method of use thereof

Abstract

[Problem] To provide a method and apparatus for simply, low-power, and automated processing of biological samples through multiple sample preparation and analysis steps. [Solution] A microfluidic apparatus comprising a reagent dispensing unit, the reagent dispensing unit comprising at least one reagent bag having one or more reagents and a frangible seal layer, and at least one plunger and at least one sharp object or projection configured to rupture the frangible seal layer and deliver one or more reagents to the microfluidic apparatus when an operating force is applied to the reagent dispensing unit. The present invention includes a microfluidic apparatus comprising a reagent dispensing unit, a sample extraction device and a sample processing unit. [Selection Diagram] Figure 1

Inventors

• パイス，アンドレア
• パイス，ロハン

Assignees

• ノベル マイクロデバイシズ，インク．

Dates

Publication Date

20260511

Application Date

20260114

Priority Date

20161201

Claims (20)

1. A microfluidic apparatus comprising a reagent dispensing unit, wherein the reagent dispensing unit is One or more reagents and at least one reagent bag having a frangible seal layer, A microfluidic device comprising at least one plunger and at least one sharp object or projection configured to rupture the frangible seal layer and deliver one or more reagents to the microfluidic device when an operating force is applied to the reagent dispensing unit.
2. The aforementioned microfluidic device further, At least one inlet conduit, At least one reagent well, Equipped with at least one waste well, The microfluidic apparatus according to claim 1, wherein the inlet conduit, the reagent well, and the waste well are fluidly connected such that an interface exists between the frangible seal and the inlet conduit, and when an operating force is applied to the reagent dispensing unit and excess reagent overflowing from the reagent well is collected in the waste well, one or more reagents are delivered to the reagent well via the inlet conduit.
3. The microfluidic apparatus according to claim 2, wherein at least two reagents are packaged in separate bags.
4. The microfluidic apparatus according to claim 2, wherein at least two reagents are packaged together in a single bag.
5. The microfluidic apparatus according to claim 4, wherein one reagent is an aqueous reagent and one reagent is a non-aqueous, miscible reagent.
6. The aqueous reagent is located closest to the interface between the frangible seal and the inlet conduit in the microfluidic apparatus according to claim 5.
7. The microfluidic apparatus according to claim 5, wherein the non-aqueous, immiscible reagent has a lower density than the aqueous reagent and floats above the aqueous reagent, thereby forming an immiscible layer on top of the aqueous reagent.
8. The microfluidic apparatus according to claim 5, wherein the aqueous reagent has a lower density than the non-aqueous, immiscible reagent and floats above the non-aqueous, immiscible reagent, thereby forming an aqueous layer on top of the non-aqueous, immiscible reagent.
9. The microfluidic apparatus according to claim 5, wherein when an operating force is applied to the reagent dispensing unit, the aqueous reagent first flows out from the inlet conduit and into the reagent well, and then the non-aqueous, immiscible reagent flows in.
10. The microfluidic apparatus according to claim 2, further comprising a locking mechanism configured to lock the plunger in the pressed position, thereby preventing backflow of the reagent into the reagent bag.
11. The microfluidic apparatus according to claim 10, wherein the locking mechanism comprises a barbed pin in a locking hole configured to restrict the movement of the plunger in a direction that facilitates pressing down the bag during the application of an operating force.
12. The microfluidic apparatus according to claim 2, comprising two or more reagent wells connected to each other and connected to one or more reagent dispensing units via primary channels.
13. The microfluidic apparatus according to claim 12, configured such that at the end of the operating sequence, the reagent wells are filled with aqueous reagents and connected to each other via the primary channels filled with a non-aqueous fluid.
14. The microfluidic apparatus according to claim 12, configured such that, at the end of the operating sequence, an immiscible oil phase is formed covering the aqueous reagent in the fluid well, and the aqueous reagent in the fluid well is separated from each other by the oil phase, but remains fluidly connected to form a fluid circuit during the sequence.
15. The microfluidic apparatus according to claim 12, comprising a plurality of reagent bags separated from the inlet conduit to the fluid well by a frangible seal, and an integrated plunger element having a locking pin that locks the plunger in the pressed position after operation, thereby preventing backflow of reagent into the reagent bags.
16. The microfluidic apparatus according to claim 15, wherein the plunger is configured to contact all of the reagent bags at the same moment and to push and release all of the reagents from the reagent bags in parallel with a single operating step.
17. The microfluidic apparatus according to claim 15, wherein the plunger is provided with spatially oriented projections having various depths, and is configured to facilitate continuous reagent delivery to the microfluidic apparatus when the plunger is pressed in contact with a desired reagent bag in a preferred sequence.
18. The microfluidic apparatus according to claim 2, further comprising a sample inlet port into which a sample can be injected.
19. The microfluidic apparatus according to claim 18, wherein the sample inlet port further comprises one or more filter membranes.
20. The microfluidic apparatus according to claim 2, further comprising a microfluidic cartridge configured to rotate between an upper actuator element and a lower actuator element, wherein the upper and lower actuator elements are equipped with spatially oriented magnets, and in a single operating step comprising rotating the microfluidic cartridge between the upper and lower actuator elements, the spatially oriented magnets capture, resuspend, and transport magnetic beads between different reagent wells.

Description

Cross-reference to related applications This application claims priority to U.S. Provisional Patent Application No. 62/428,976, filed on 1 December 2016, which is incorporated herein by reference in its entirety. This invention relates to an automated point-of-care apparatus for complex sample processing and a method for using the same. Point-of-care (POC) devices enable convenient and rapid testing in patient care settings. Therefore, sample-to-answer and lab-on-a-chip (LOC) systems, which integrate microfluidic technology, are becoming increasingly popular. These LOCs integrate various laboratory functions, such as extraction, amplification, detection, interpretation, and reporting, previously performed manually and/or off-site, all onto a single device. Because sample-to-answer and LOC testing is conducted in patient care settings rather than in laboratory facilities, these types of testing have presented contamination control issues, particularly in processes involving human interaction during processing. Therefore, there is a need to automate sample handling within sample-to-answer LOCs to minimize human interaction. These sample-to-answer and LOCs are typically several square millimeters to several square centimeters in size and are often of the micro-electromechanical system (MEMS) type. Here, MEMS capable of detecting and analyzing biological substances are generally called Bio-MEMS. Most POC diagnostic devices on the market are classified as highly or moderately complex under the Clinical Laboratory Improvement Amendments (CLIA). These federal guidelines generally apply to human clinical laboratory testing devices, with the exception of certain conditions that exempt them. One of these conditions is that the device or equipment meets specific risk, error, and complexity requirements. To qualify a POC diagnostic test for CLIA exemption, sample preparation and fluid handling steps must be minimized. One way to minimize these steps is to store reagents in sealed devices such as openable blister or burst bags. Reagent delivery to microfluidic chips generally involves the use of pumps such as syringe pumps or peristaltic pumps, and bottles, syringes, or reservoirs filled with external reagents. These systems are complex not only because they are difficult to carry, but also because they require numerous components that must be integrated and a leak-free fluid interface to the microfluidic chip. The latest technologies on the market have yet to successfully implement a method for easily automating fluid handling in a compact and low-power manner. Consequently, this has been seen as an obstacle to conducting proof-of-concept (POC) studies in the vast majority of multi-stage biological assays still performed in large clinical settings. Complex biological assays requiring multiple processing steps, including but not limited to pipetting, heating, cooling, mixing, washing, culturing, labeling, binding, and elution, rely on expensive laboratory automation equipment to perform sample-to-answer sequencing. Low-cost, low-power, and miniaturized instrumentation for automating sample-to-answer sequencing has not yet been realized; therefore, point-of-care microfluidic systems for performing sample-to-answer sequencing rely on additional instrumentation in the form of standalone benchtop or portable devices for performing analysis on the microfluidic system. Implementing separate instrumentation capable of automating the sample preparation process on microfluidic cartridges has been considered a way to keep the cost per test, and therefore the cost of the cartridge, low. Systems developed for point-of-care applications can take the form of portable benchtop devices equipped with solenoid plungers, linear actuators, microcontrollers, and electronic circuits to automate the sample preparation sequence. This instrumentation configuration allows user control of the sample processing sequence, but requires a controlled environment and a significant amount of power to operate. These point-of-care systems are not feasible in low-resource environments where the infrastructure for operating the instruments is lacking, or in non-hospital settings such as homes where laypeople do not need, cannot afford, or are not trained to operate expensive instruments for testing. Therefore, developing methods to enable low-power, standalone, inexpensive, and disposable instrumentation configurations that can be directly integrated onto microfluidic systems and perform automated sample-to-answer sequencing is considered an obstacle to developing single-use test devices capable of performing complex multi-step nucleic acid, protein, and immunological assays from sample-to-answer. Disposable tests that do not require instrumentation to perform them are limited to simple single-step and multi-step analyses. In simple single-step analyses, the sample is a single liquid, and no reagents are used. These tests typically include dipstick tests su