Digital molecular assays
AssigneeIlytica LLC
InventorJay T. Groves
Life of this patent
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Abstract
Provided herein are systems, devices and methods for the rapid and accurate measurement of analytes by assay of binding events, by direct, digital measurement of individually resolved analyte/reporter binding events. The digital molecular assay systems, devices and methods disclosed herein are capable of particle-by-particle readout using optical reporter molecules that detect and report the binding of a single analyte molecule, and report each such binding in binary format. Such digital molecular assay systems, devices and methods are useful in a variety of applications, such as on mobile electronic devices for use in the field.
Claims42
A method for determining presence or concentration of at least one analyte in a sample, comprising:
- in an image of a plurality of signals emitted by at least one type of optical reporter molecules incubated with at least one type of analyte molecules,
- for each type of optical reporter molecules, determining in the image which optical reporter molecules are active and which are null;
- determining from the active optical reporter molecules the number bound to analyte molecules and the number of active optical reporter molecules unbound to analyte in the image by individually resolving bound and unbound active optical reporter molecules;
- and
- determining the presence or concentration of analyte from the number of bound active optical reporter molecules as a fraction of, or as proportional to a fraction of, the total number of bound and unbound active optical reporter molecules.
A method for determining presence or concentration of antigen in a sample, comprising:
- in an image of a plurality of signals emitted by at least one type of optical reporter molecules comprising antibodies functionalized onto one or more plasmonic nanoparticles incubated with antigen,
- for each type of optical reporter molecule, determining in the image which possess active antibodies and which possess null antibodies;
- determining the number of active antibodies bound to antigen and the number of active antibodies unbound to antigen in the image by individually resolving bound and unbound optical reporter molecules;
- and
- determining the presence or concentration of antigen from the number of bound active antibodies as a fraction of, or as proportional to a fraction of, the total number of bound and unbound active antibodies.
A method for determining presence or concentration of a target nucleotide sequence in a sample, comprising:
- in an image of a plurality of signals emitted by
- a) an optical reporter molecule comprising a first capture nucleotide sequence complementary to a first part of the target nucleotide sequence and
- b) bound complexes comprising a target nucleotide sequence bound to the first optical reporter molecule comprising the first capture nucleotide sequence complementary to part of the target nucleotide sequence and a second optical reporter molecule comprising a second capture nucleotide sequence complementary to a second part of the target nucleotide sequence,
- determining in the image which optical reporter molecules are active and which are null;
- determining the number of active bound complexes, comprising target nucleotide sequences bound to active optical reporter molecules comprising the first and second parts of the complementary nucleotide sequence;
- determining the presence or concentration of the target nucleotide sequence as a fraction of, or as proportional to number of active bound complexes as a fraction of the total number of active optical reporter molecules emitting detectable signals.
Dependencies are inferred from the claim text.