Biomarker Detection From Fluid Samples
AssigneeThe Board of Trustees of the University of Illinois
InventorsRashid BASHIR, Anurup GANGULI
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Abstract
Described herein are systems and methods which utilize an array of wells to isolate pathogens and nucleic acid detection techniques to accurately and rapidly detect pathogens in fluid samples, even in very low concentrations, including from solid or semi-solid samples that have been fluidized. The provided systems and methods dry the fluid sample to deposit a fraction of the total volume in a number of wells and perform nucleic acid detection on individual wells to detect even individual pathogens and provide a quantitative analysis of the amount of pathogen within the sample. Also provided are methods and systems for precise delivery of dried materials, including biomolecules that are enzymes of use in the process, to microwells.
Claims40
A method of detecting one or more target analytes from a liquid sample, the method comprising the steps of:
- applying a liquid sample to a substrate, wherein the substrate has an array of wells;
- drying the liquid sample to generate a plurality of dried sample islands, with each sample island confined to a unique well;
- applying a reagent to each well, wherein the reagent is used for nucleic acid detection;
- eluting a nucleic acid from the dried sample islands with a liquid phase having the applied reagent;
- detecting a target nucleic acid if present in a dried sample island by a bi-phasic reaction, wherein the detecting is by electrical or optical detection;
- thereby detecting the one or more target analytes from the liquid sample.
A system for introducing a biomolecule to a plurality of wells in an array for target analyte detection, the system comprising:
- a plate having a plurality of microwells, each microwell having:
- a well surface;
- a top portion that is physically accessible;
- and
- wherein adjacent microwells are separated by a separation surface;
- a hydrophobic substrate having a release surface;
- a biomolecule that is connected to the hydrophobic substrate release surface;
- wherein the hydrophobic substrate release surface with biomolecules connected thereto is configured to connect to the microwell separation surface and fluidically seal the microwells to prevent fluid transmission between different microwells filled with a fluid.
Dependencies are inferred from the claim text.