Multivolume devices, kits and related methods for quantification and detection of nucleic acids and other analytes

• US 9,493,826 B2
• Filed: 02/10/2014
• Issued: 11/15/2016
• Est. Priority Date: 03/24/2009
• Status: Active Grant

• Current Assignee: University of Chicago
• Original Assignee: California Institute of Technology, University of Chicago
• Inventors: Ismagilov, Rustem F., Shen, Feng, Kreutz, Jason E., Du, Wenbin, Sun, Bing
• Primary Examiner(s): HURST, JONATHAN M
• Application Number:
  US14/177,190
• Publication Number:
  US 20140336064A1
• Time in Patent Office: 1,009 Days
• US Class Current: 1/1
• CPC Class Codes: B01F 25/14 Mixing drops, droplets or bodies of liquid which flow together or contact each other micromixers using where the materials to be mixed are in the form of droplets B01F33/302
• CPC Class Codes: B01F 33/304 the mixing being performed in a mixing chamber where the products are brought into contact
• CPC Class Codes: B01F 35/71411 rotating or oscillating about an axis
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2300/045 whereby the whole cover is slidable
• CPC Class Codes: B01L 2300/049 Valves integrated in closure
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 2400/065 sliding valves
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 2531/119 Strand displacement amplification [SDA]
• CPC Class Codes: C12Q 2537/149 Sequential reactions
• 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
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device

Abstract

Provided are devices comprising multivolume analysis regions, the devices being capable of supporting amplification, detection, and other processes. Also provided are related methods of detecting or estimating the presence nucleic acids, viral levels, and other biological markers of interest.

Claims

1. A method of binary quantification of nucleic acids in a sample, comprising:

a. providing a microfluidic device comprising a first substrate comprising a first population of areas and a second substrate comprising a second population of areas;

b. partitioning a sample comprising nucleic acid molecules into said first and second populations of areas in said microfluidic device by exerting the sample through a conduit in fluidic communication with each area of the first population of areas and each area of the second population of areas via a continuous fluidic pathway within said first and second substrates, such that a plurality of said first and second populations of areas each comprise only one nucleic acid molecule;

c. loading nucleic acid amplification reagents into said microfluidic device;

d. effecting relative motion between the first substrate and the second substrate, thereby isolating the first population of areas from the second population of areas; and

e. conducting a nucleic acid amplification reaction in said microfluidic device in both said first and second populations of areas, wherein during said nucleic acid amplification reaction the temperature of said sample and the temperature of said nucleic acid amplification reagents remain within 10°

C. of their temperature at the beginning of said nucleic acid amplification reaction.

2. The method of claim 1, wherein said nucleic acid amplification reaction comprises loop mediated isothermal amplification.

3. The method of claim 1, wherein said nucleic acid amplification reaction comprises reverse transcription loop mediated isothermal amplification.

4. The method of claim 1, wherein said nucleic acid amplification reaction comprises nucleic acid sequence based amplification.

5. The method of claim 1, wherein said nucleic acid amplification reaction comprises sequence-specific polymerase chain reaction.

6. The method of claim 1, wherein said nucleic acid amplification reaction comprises transcription mediated amplification.

7. The method of claim 1, wherein said nucleic acid amplification reaction comprises transcription associated amplification.

8. The method of claim 1, wherein said nucleic acid amplification reaction comprises reverse transcription recombinase polymerase amplification.

9. The method of claim 1, wherein said nucleic acid amplification reaction comprises strand displacement amplification.

10. The method of claim 1, wherein said nucleic acid amplification reaction comprises signal mediated amplification of RNA technology.

11. The method of claim 1, wherein said nucleic acid amplification reaction comprises cyclic enzymatic amplification method.

12. The method of claim 1, wherein said nucleic acid amplification reaction comprises isothermal target and signaling probe amplification.

13. The method of claim 1, wherein said nucleic acid amplification reaction comprises ligase chain reaction.

14. The method of claim 1, wherein said microfluidic device comprises a heat control component.