Digital assay for quantifying and concentrating analytes

Patent metadata

Field	Value
Publication number	US-2016266105-A1
Application number	US-201615000864-A
Country	US
Kind code	A1
Filing date	Jan 19, 2016
Priority date	Jul 19, 2013
Publication date	Sep 15, 2016
Grant date	—

Abstract

Provided herein are methods, compositions, and devices for the identification and quantification of analytes, such as nucleic acids, proteins, cells or other biological samples. The methods, compositions, and devices are suited for accurate, portable analysis of small amounts of analyte.

First claim

1 . A method for analyzing a sample, comprising:
providing a device comprising
a fluid inlet,
a fluid receiving structure,
a fluid path connecting said fluid inlet and said receiving structure, and
a plurality of digital units located in said fluid path, each digital unit capable of capturing at least one analyte;
flowing a fluid sample in said fluid path, said fluid sample comprising analytes; capturing a subset of said analytes with said digital units; isolating a first subset of said digital units from a second subset of said digital units; and determining the concentration or identity of said analytes in said fluid sample performing digital amplification of the analytes.

2 . The method of claim 1 , wherein said determining is conducted based on signal generated from said digital units.

3 . The method of claim 2 , wherein signal is generated from each of said digital units if the number of analytes captured thereby exceeds a threshold number.

4 . The method of claim 1 , wherein isolating the first subset of said digital units from the second subset of said digital units comprises actuating a portion of the device.

5 . The method of claim 3 , wherein said threshold number is greater than zero.

6 . The method of claim 2 , wherein the probability of signal being generated from each of said digital units depends of the number of analytes captured thereby.

7 . The method of claim 6 , wherein said probability is different between a first digital unit of said plurality of digital units and a second digital unit of said plurality of digital units.

8 . The method of claim 7 , wherein a first digital unit of said plurality of digital units and a second digital unit of said plurality of digital units are exposed to different amounts of sample.

9 . The method of claim 2 , wherein said determining comprises generating a profile of said signal versus location along said flow path and comparing said profile to a calibration profile.

10 . The method of claim 2 , wherein said determining comprises measuring the number of said digital units after which said signal decreases below a threshold value.

11 . (canceled)

12 . The method of claim 2 , wherein said determining comprises comparing signal from said first subset of said digital units to signal from said second subset of said digital units.

13 . The method of claim 1 , wherein said isolating is achieved by physical motion of said digital units.

14 . The method of claim 1 , wherein said isolating is achieved by introduction of an isolation fluid into said fluid path.

15 . The method of claim 1 , wherein said flowing comprises flow through said digital units.

16 .- 19 . (canceled)

20 . The method of claim 1 , wherein, prior to said determining, said analytes are released from said digital units.

21 .- 25 . (canceled)

26 . A device for analyzing a sample comprising analytes, comprising:
a fluid inlet, a fluid receiving structure, a fluid path connecting said fluid inlet and said fluid receiving structure, a plurality of digital units comprising a first subset of digital units and a second subset of digital units configured to be physically isolatable from the first subset, wherein each digital unit is capable of capturing said at least one analyte; and a signal generator for generating signal from said digital units by digital amplification.

27 . The device of claim 26 , wherein said signal generator generates a signal from each of said digital units if the number of analytes bound thereto exceeds a threshold number.

28 . (canceled)

29 . The device of claim 27 , wherein said threshold number is greater than zero.

30 . The device of claim 27 , wherein said threshold number for a first digital unit of said digital units is different than said threshold number for a second digital unit of said digital units.

31 . The device of claim 26 , wherein the number of digital units from which signal is generated is correlated to the amount of said analytes in said sample.

32 . The device of claim 26 , wherein the number of digital units along said flow path after which said signal decreases below a threshold value is correlated to the amount of said analytes in said sample.

33 . (canceled)

34 . The device of claim 26 , wherein at least part of said sample flows through said digital units.

35 . The device of claim 26 , wherein the smallest dimension of each of said digital units is less than or equal to about 1 mm.

36 .- 38 . (canceled)

39 . The device of claim 26 , wherein said receiving structure comprises a hydrophobic element.

40 . (canceled)

Assignees

• California Inst Of Techn
• Univ Chicago

Inventors

• Ismagilov Rustem F
• Mccalla Stephanie
• Schlappi Travis
• Huynh Toan
• Rolando Justin
• Liu Weishan
• Ge Shencheng
• Kreutz Jason E

Classifications

• B01L2300/0893

  having a very large number of wells, microfabricated wells · CPC title

• B01L2300/0819

  Microarrays; Biochips · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• B01L2300/0803

  Disc shape · CPC title

• B01L2300/12

  Specific details about materials · CPC title