Device for detection of target molecules and uses thereof
Inventors
Siciliano, Nicholas A. • Bouliane, Martin Joseph
Assignees
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Publication Number
US-9816984-B2
Publication Date
2017-11-14
Expiration Date
Abstract
Devices and methods for the detection of antigens are disclosed. Devices and methods for detecting food-borne pathogens are disclosed.
Core Innovation
The invention relates to an antigen detection membrane system for food-borne pathogens, where a vertical-flow chromatographic membrane stack is arranged with a conjugate pad, a permeable membrane, a test membrane, and two absorbent members. The membrane detection system is housed in a housing having a first housing member, a second housing member, and an inlet opening in fluid contact with the membrane detection system.
A force member applies pressure substantially perpendicular to the membrane detection system, with the vertical axis of the force member substantially perpendicular to the membrane detection system. A slidable locking member contacts the force member and is operably connected to the conjugate pad so that moving the slidable locking member moves the conjugate pad. The sample flows vertically from the conjugate pad to the test membrane, and moving the conjugate pad after a portion of the sample has contacted and flowed through the conjugate pad exposes at least a portion of the test membrane for detection.
The device and method support identifying a positive or negative reaction for a target molecule, including detection formats using capture reagents on the conjugate pad and the test membrane, and using label options such as colloidal gold. An inlet/sample handling system and kit components are described, and an optional hydrophobic membrane is included to modulate dwell time, with capture reagents and pathogen targets such as E. coli 0157 LPS described in an example assay.
Claims Coverage
The independent claim covers detecting a target molecule using a membrane detection system inside a device, with a vertically flowing sample path, perpendicular compression via a force member, and a slidable locking mechanism that moves the conjugate pad to expose the test membrane for detection, followed by identifying a positive or negative reaction. The coverage includes multiple refinements through dependent claims defining structural relationships and biological capture reagent binding targets and example pathogen targets.
Vertical-flow membrane detection system with parallel planar alignment
A housing with an inlet opening and a membrane detection system in which a conjugate pad comprising a first capture reagent, a permeable membrane, a test membrane comprising a second capture reagent, and two absorbent members are substantially parallel to each other and the inlet opening, with the sample flowing vertically from the conjugate pad to the test membrane.
Perpendicular compression with force member aligned to membrane detection system
A force member applies pressure substantially perpendicular to the membrane detection system, wherein the vertical axis of the force member is substantially perpendicular to the membrane detection system.
Slidable locking mechanism that moves the conjugate pad to expose the test membrane
A slidable locking member contacts the force member, with an attachment member in contact with the slidable locking member and a sliding button attached to the slidable locking member, wherein moving the slidable locking member moves the conjugate pad after a portion of the sample has contacted and flowed through the conjugate pad, thereby exposing at least a portion of the test membrane for detection.
Positive or negative identification of target molecule
The method identifies a positive or negative reaction for the target molecule.
Capture reagent binding to polynucleotide, peptide, protein, saccharide, or carbohydrate
The first capture reagent binds to a polynucleotide, a peptide, a protein, a saccharide, or a carbohydrate.
Antibody as first capture reagent
The first capture reagent is an antibody.
Labeled first capture reagent options for detection
The first capture reagent includes colloidal gold, a fluorescent molecule, a radioactive tag, an infrared molecule, or a chemiluminescent substrate.
Example pathogen protein or polynucleotide targets
The pathogen protein or pathogen polynucleotide is from E. coli, Campylobacter, Listeria, or Salmonella.
The claim set is directed to a vertical-flow membrane detection system combined with perpendicular compression and a slidable locking member that moves the conjugate pad after initial flow to expose the test membrane, enabling identification of a positive or negative reaction. Dependent claims further define the first capture reagent binding scope, including antibody and labeled options, and provide example pathogen targets.
Stated Advantages
Provides a mechanism to expose at least a portion of the test membrane for detection after a portion of the sample has contacted and flowed through the conjugate pad.
Enables identification of a positive or negative reaction for the target molecule.
Documented Applications
Detection of food-borne pathogens using an antigen detection membrane system, including detection of E. coli 0157 LPS in an example assay format.
Detection targets include Campylobacter and Salmonella as part of the described example pathogen scope.
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