RARE CELL ANALYSIS AFTER NEGATIVE SELECTION

• US 20140303005A1
• Filed: 04/04/2014
• Published: 10/09/2014
• Est. Priority Date: 04/05/2013
• Status: Abandoned Application

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Raindance Technologies Incorporated (Bio-Rad Laboratories Incorporated)
• Inventors: Samuels, Michael, Link, Darren R.
• Application Number:
  US14/245,348
• Publication Number:
  US 20140303005A1
• Time in Patent Office: Days
• US Class Current: 506/2
• CPC Class Codes: C12N 15/1075 by coupling phenotype to genotype, not provided for in other groups of this subclass
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/159 Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components

Abstract

The invention generally relates to rare cell analysis after negative selection.

Claims

1. A method for detecting biological material from a target cell, the method comprising:

obtaining a sample suspected to contain a target cell;

conducting an assay on the sample that removes non-target cells from the sample;

extracting biological material from the remaining target and non-target cells; and

analyzing the biological material using a digital counting technique, thereby detecting the biological material from a target cell.

2. The method according to claim 1, wherein the assay results in the target cell being present at about or less than 1 in 1,000,000 cells.

3. The method according to claim 1, wherein the assay results in the target cell being present at about or less than 1 in 10,000 cells.

4. The method according to claim 1, wherein the biological material is nucleic acid.

5. The method according to claim 4, wherein the digital counting technique is selected from the group consisting of is digital PCR and sequencing.

6. The method according to claim 5, wherein sequencing is sequencing-by-synthesis.

7. The method according to claim 1, wherein the biological material is protein.

8. The method according to claim 7, wherein the digital counting technique is digital ELISA.

9. The method according to claim 1, wherein analyzing comprises:

compartmentalizing the extracted biological material into compartmentalized portions; and

conducting an assay on the biological material in each of the compartmentalized portions.

10. The method according to claim 9, wherein the compartmentalized portions are aqueous droplets in an immiscible fluid.

11. The method according to claim 10, wherein the immiscible fluid is oil.

12. The method according to claim 11, wherein the oil is a fluorinated oil.

13. The method according to claim 11, wherein the oil comprises a surfactant.

14. The method according to claim 13, wherein the surfactant is a fluorinated surfactant.

15. The method according to claim 10, wherein the droplets are flowing through a channel.

16. The method according to claim 15, wherein the biological material is nucleic acid and the assay comprises amplifying the nucleic acid within the compartmentalized portions.

17. The method according to claim 16, wherein a plurality of the droplets comprises no more than a single target nucleic acid in each droplet.

18. The method according to claim 9, wherein the compartmentalized portions are produced by a flow focusing technique.

19. The method according to claim 18, wherein the biological material is in an aqueous solution and the compartmentalized portions are produced by partitioning the aqueous fluid with an immiscible fluid.

20. The method according to claim 19, wherein the aqueous fluid is flowing through a channel.