Methods of in situ detection of nucleic acids
AssigneeAdvanced Cell Diagnostics, Inc.
InventorsYuling Luo, Huei-Yu Wang, John James Flanagan, Shiping Chen
Life of this patent
Estimating the term…
Abstract
Methods of detecting the presence or absence of a class of nucleic acid targets in single cells through direct or indirect capture of labels to the nucleic acids are provided, where such labels to the class of nucleic acid targets are indistinguishable from each other. Also described are methods of detecting individual cells, particularly a cell of a specific type from large heterogeneous cell populations, through detection of one or more of nucleic acid targets, where the labels to the one or more of nucleic acid targets are indistinguishable from each other. Related kits are also described.
Claims24
A method of detecting an individual cell of a specified type, the method comprising:
- providing a sample comprising a mixture of cell types, which mixture comprises at least one cell of the specified type;
- providing a first label probe comprising a first label, and providing a second label probe comprising a second label, wherein a first signal from the first label is indistinguishable from a second signal from the second label;
- capturing, in the cell, the first label probe to a first nucleic acid target, when present in the cell, and the second label probe to a second nucleic acid target, when present in the cell;
- detecting the signal from the labels;
- correlating the signal detected from the cell with the presence, absence, or amount of the first or second nucleic acid targets in the cell;
- and
- identifying the cell as being of the specified type based on detection of the presence, absence, or amount of either the first or second nucleic acid targets within the cell,
- wherein the specified type of cell is distinguishable from the other cell type(s) in the mixture on the basis of either the presence, absence, or amount of the first nucleic acid target or the presence, absence, or amount of the second nucleic acid target in the cell.
11-31. (canceled)
A kit for detecting an individual cell of a specified type, comprising:
- at least a first capture probe capable of hybridizing to the first nucleic acid target;
- at least a second capture probe capable of hybridizing to the second nucleic acid target;
- a first label probe comprising a first label and a second label probe comprising a second label, wherein the first label probe is capable of hybridizing to the first capture probe and the second label probe is capable of hybridizing to the second capture probe;
- wherein a first signal from the first label is indistinguishable from a second signal from the second label;
- wherein the specified type of cell is distinguishable from the other cell types in the mixture by presence, absence, or amount of the first nucleic acid and/or the second nucleic acid target;
- and
- packaged in one or more containers.
36-54. (canceled)
A method of detecting the presence or absence of a class of nucleic acid targets in an individual cell, wherein said class of nucleic acid targets consists of a plurality of nucleic acid targets, the method comprising:
- providing a sample comprising the cell, which cell comprises or is suspected of comprising said class of nucleic acid targets;
- providing, for each of the plurality of nucleic acid targets, a label probe comprising a label, wherein a signal from the label of one nucleic acid target in the plurality of nucleic acid targets is distinguishable from a signal from the label of another nucleic acid target in the plurality of nucleic acid targets;
- providing, for each of the plurality of nucleic acid targets, at least a capture probe;
- hybridizing in the cell, for each of the plurality of nucleic acid targets, the capture probe to its corresponding nucleic acid target, when present in the cell;
- capturing, for each of the plurality of nucleic acid targets, the label probe to the capture probe, thereby capturing the label probe to its corresponding nucleic acid target;
- and
- detecting the signal from the label of the label probes captured to the plurality of nucleic acid targets.
57-59. (canceled)
64-75. (canceled)
Dependencies are inferred from the claim text.