Assay for detection of androgen receptor variants
AssigneeCornell University
InventorsParaskevi Giannakakou, Ada Gjyrezi, Seaho Kim
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Abstract
Sensitive assay methods are described herein that involve (a) capturing circulating cancer cells (e.g., by any method); (b) extracting mRNA and (c) detecting and quantifying each of full-length androgen receptor (AR-FL), androgen receptor variant 7 (AR-V7), and androgen receptor variant 5,6,7 (AR-V567) in parallel digital droplet PCR assays. Additional methods are described herein for detecting and/or isolating circulating cancer cells by selecting for cells that express transferrin receptor.
Claims31
A method comprising:
- a. capturing circulating cancer cells from a sample from a test subject;
- b. extracting mRNA to provide a RNA sample;
- and
- c. detecting and quantifying each of full-length androgen receptor (AR-FL), androgen receptor variant 7 (AR-V7), and androgen receptor variant 5,6,7 (AR-V567) in parallel digital droplet PCR assays;
- wherein the parallel digital droplet PCR assays separately comprise mixtures of two or more primers or probes comprising sequences SEQ ID NO:7, 8, 9, 11, 12, 13, 15, 16, or 17.
A composition comprising two or more types of primers or probes with at least 15 nucleotides of SEQ ID NO:7, 8, 9, 11, 12, 13, 15, 16, and 17, wherein one or more type of primer or probe is covalently or non-covalently bound to a label.
The composition of statement 24, further comprising a probe comprising at least 15 nucleotides of a SEQ ID NO:13 sequence.
A method comprising:
- a. obtaining a sample from a test subject;
- b. depleting the sample of CD45+ cells to obtain a CD45+ depleted sample;
- c. staining the CD45+ depleted sample with a labeled reagent that binds to transferrin receptor to obtain a TfR-labeled sample;
- d. optionally quantifying the TfR-labeled cells in the TfR-labeled sample;
- e. optionally measuring RNA or protein expression in the TfR-labeled sample.
Dependencies are inferred from the claim text.