Rotary manifold for paper-based immunoassays
Inventors
Henry, Charles S. • FEENY, Rachel • FRANKLIN, Alan B. • CARRELL, Cody
Assignees
US Department of Agriculture USDA • Colorado State University Research Foundation
Publication Number
US-11291997-B2
Publication Date
2022-04-05
Expiration Date
2039-08-02
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Abstract
A system for and methods of analyzing a test sample through the use of a rotary apparatus that includes a microfluidic paper-based apparatus (mPAD). The apparatus includes two or more layers that are rotatable with respect to one another. A middle layer may comprise a microfluidic apparatus having one or more reagent channels. Each of the reagent channels may include reagent dried on the surface of the channel, and, together with an absorption pad, may be aligned vertically with a sample chamber. Male and female engagement surfaces on each of the middle layer, the top layer, and the bottom layer interlock to secure each layers in vertical alignment so that fluid flows through the apparatus to contact a test sample with a reagent and facilitate detection of a target analyte in the test sample in the sample chamber.
Core Innovation
The invention provides a system and methods for analyzing a test sample using a rotary microfluidic paper-based apparatus comprising a housing with multiple layers, including a rotatable middle microfluidic layer. This middle layer contains reagent channels with dried reagents and absorption pads arranged radially about a central vertical rotational axis. A sample chamber in a removable sample insert is located in the bottom layer. The rotation of the middle layer aligns a reagent channel and absorption pad vertically with the sample chamber, enabling fluid flow and contact between the test sample and reagents for analyte detection.
The apparatus is designed with integral male and female engagement surfaces on the top, middle, and bottom layers that interlock to secure vertical alignment and restrict rotation to defined increments. This construction allows sequential delivery of various reagents and wash buffers to the sample chamber. The housing can be fabricated via three-dimensional printing or traditional molding, providing customizable and reproducible assembly for sensitive and controlled diagnostic assays.
Claims Coverage
The patent includes two independent claims defining the microfluidic apparatus and its method of analyzing a test sample, focusing on the multi-layer rotary construction, reagent delivery, fluid communication, and engagement surfaces for alignment and rotation control.
Rotary multi-layer housing with integral engagement surfaces
The apparatus comprises a housing with top, middle, and bottom substantially planar layers oriented perpendicular to a central vertical rotational axis. The middle layer is rotatable relative to the top and bottom layers and includes plural integral male and female engagement surfaces that interlock complementarily with those on the other layers to secure vertical alignment and permit controlled rotation.
Removable microfluidic layer with radially arranged reagent channels and absorption pads
The middle layer contains a removable microfluidic layer including multiple reagent channels and absorption pads arranged radially about the central vertical rotational axis, with reagents dried on the surfaces of the reagent channels to enable sequential reagent delivery to the sample chamber.
Fluid communication via aligned reagent channels, absorption pads, and sample chamber
Rotation of the middle layer vertically aligns a reagent channel and absorption pad with the fluid reservoir and sample chamber in the bottom layer, establishing fluid communication between these components to facilitate analyte detection in the test sample.
Defined rotation increments via engagement surface arrangement
The male and female engagement surfaces of the middle, top, and bottom layers are arranged to permit rotation of the middle layer relative to the other layers in fixed increments, including specific embodiments with 45-degree rotational steps or increments ranging from about 10 degrees to about 180 degrees.
The independent claims cover the invention of a rotatable, multi-layer paper-based microfluidic apparatus with interlocking engagement surfaces for precise reagent delivery through vertically aligned reagent channels to a sample chamber, enabling controlled sequential assays with customizable rotation increments.
Stated Advantages
Provides improved consistency of vertical alignment between reagent channels and the sample chamber through interlocking male and female engagement surfaces.
Allows sequential delivery of various reagents and wash buffers to a test sample, facilitating complex multi-step diagnostic assays.
Includes a disposable microfluidic paper-based device combined with a low-cost, portable, reusable housing potentially fabricated by three-dimensional printing.
Enables customizable absorbent pads to control fluid flow through the apparatus, improving assay performance.
Supports easy swapping of capture molecules to detect different analytes, enhancing assay flexibility.
Documented Applications
Point-of-care diagnostic assays for detection of analytes such as pathogens (e.g., Salmonella), proteins, small molecules, nucleic acids, and other biomarkers in biological samples.
Immuno-magnetic separation sandwich immunoassays using magnetic beads coated with capture molecules to isolate and detect target analytes in test samples.
Colorimetric detection of bacterial contamination in media or milk using sequential reagent delivery in the apparatus.
Use in resource-limited settings for rapid and sensitive diagnostic testing without laboratory infrastructure.
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