Methods for detecting multiple analytes with a single signal
Inventors
Siciliano, Nicholas • Leong, Louis • Keough, Martin Patrick • Brown, Ashley Shaniece
Assignees
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Publication Number
US-9347938-B2
Publication Date
2016-05-24
Expiration Date
Abstract
Compositions, methods, and devices for the detection of multiple analytes with a single signal are provided.
Core Innovation
The invention provides methods and devices for detecting analytes with a single signal by using a bridging complex that forms only when multiple analytes of interest are present. A first analyte of interest and a second analyte of interest are different, and the signal detection unit binds to the bridging complex to generate the single signal indicative of concurrent presence or absence of the analytes.
In the device aspect, a housing includes a force member and a detection membrane system comprising a conjugate pad, a test membrane, and an absorbent member, with an inlet opening in fluid contact with the conjugate pad. Pressure applied substantially perpendicular to the detection membrane system and an arrangement of the conjugate pad, test membrane, and absorbent member substantially parallel enable the sample to contact the membrane system so that the single-signal detection can be performed when the bridging complex is formed.
The document also describes a vertical-flow multi-analyte detection approach with single-signal readout, including actuation and a single optical signal detection workflow using an optical reader or spectrometer with signal processing and a display unit. Example embodiments show bridging-complex detection of two PCR amplicon analytes using streptavidin-bridge formation logic, where signal is generated only when the first analyte, second analyte, and bridge unit form the bridging complex, and a bridging workflow using labeled amplicons and a streptavidin-gold conjugate pad, where colloidal gold signal appears only when the bridging complex forms.
Claims Coverage
The document includes three independent claims: a two-analyte single-signal bridging detection method, a three-analyte single-signal bridging detection method, and a concurrent plurality-of-analytes single-signal detection method/device configuration. Across these claims, the core coverage centers on generating a single signal only when specified analytes and bridge units form a bridging complex that is bound by a signal detection unit (capture reagent).
Concurrent detecting of first and second analytes with a single signal via bridging complex binding
The method concurrently detects a first analyte of interest and a second analyte of interest in a test sample with a single signal, wherein the first and second analytes from the sample are different, the single signal indicates presence or absence concurrently, and the single signal is detected only when the first analyte, the second analyte, and a bridge unit form a bridging complex and the signal detection unit binds to the bridging complex.
Concurrent detecting of first, second, and third analytes with a single signal via bridging complex binding
The method concurrently detects a first analyte of interest, a second analyte of interest, and a third analyte of interest in a test sample with a single signal, wherein the first, second, and third analytes are different, and the single signal is detected only when the first, second, third analytes and a first bridge unit and a second bridge unit form a bridging complex and the signal detection unit binds to the bridging complex.
Concurrent detecting of a plurality of analytes with a single-signal device using a detection membrane system and pressure perpendicular to the membrane
The method concurrently detects a plurality of analytes in a test sample with a single signal by contacting a device comprising a housing with a force member, a slidable locking member, an attachment member, a sliding button, and a detection membrane system having a conjugate pad, a test membrane, and an absorbent member arranged substantially parallel; the force member applies pressure substantially perpendicular to the detection membrane system and the sample contains a first analyte, a second analyte, and a bridge unit, with a signal detection unit comprising a third capture reagent that binds based on formation of a bridging complex so that the single signal is detected only when the first analyte, second analyte, and bridge unit form the bridging complex.
Overall, the independent claims cover bridging-complex based concurrent multi-analyte detection in which a single signal is generated only upon formation of a bridging complex from specified analytes and bridge units, followed by binding of the bridging complex by a signal detection unit that includes a capture reagent.
Stated Advantages
Avoidance of false positives by detecting the single signal only when the complex contains the first and second analytes.
Documented Applications
Food-borne pathogen detection including Shiga toxin, eae gene, and E. coli serotypes.
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