Multivolume devices, kits and related methods for quantification and detection of nucleic acids and other analytes
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.
1. A method, comprising:
a) introducing a sample comprising a target molecule into a device comprising
a first component comprising a plurality of first areas,
a second component comprising a plurality of second areas,
wherein the first and second components are configured to slip relative to the other between
a first position, wherein the plurality of first areas and the plurality of second areas overlap to form a continuous fluidic path, and
a second position, wherein at least two areas of said plurality of first areas and second areas are isolated from each other;
b) in said first position, distributing an amount of said target molecule into said at least two areas of the plurality of first areas and second areas via the continuous fluidic path extending through the overlapping plurality of first areas and second areas and which connects the at least two areas, wherein said at least two areas define volumes that differ from one another;
c) slipping said first or second component relative to the other to the second position, thereby isolating the at least two areas;
(d) effecting a reaction on said amount of said target molecule in said at least two isolated areas and thereby producing a reaction product in said at least two isolated areas;
(e) detecting said reaction product optically in said at least two areas; and
(f) estimating, from said reaction product, a level of said target molecule in said sample.
2. The method of claim 1 , wherein said target molecule comprises a nucleic acid.
3. The method of claim 2 , wherein effecting the reaction comprises contacting an amplification reagent with said nucleic acid.
4. The method of claim 2 , wherein at least one of said at least two areas is estimated to comprise about one molecule of nucleic acid.
5. The method of claim 1 , wherein at least one of said at least two areas is estimated to contain only target molecule.
6. The method of claim 1 , wherein said reaction comprises nucleic acid amplification.
7. The method of claim 6 , wherein said nucleic acid amplification comprises polymerase chain reaction, room-temperature polymerase chain reaction, nested polymerase chain reaction, multiplex polymerase chain reaction, arbitrarily primed polymerase chain reaction, nucleic acid sequence-based amplification, transcription mediated amplification, strand displacement amplification, branched DNA probe target amplification, ligase chain reaction, cleavase invader amplification, anti DNA-RNA hybrid antibody amplification, or any combination thereof.
8. The method of claim 6 , wherein said nucleic acid amplification is essentially isothermal.
9. The method of claim 1 , wherein at least one of said at least two areas defines a volume in the range of from about 1 picoliter to about 1 microliter.
10. The method of claim 1 , wherein distribution comprises effecting relative motion between the first and second component so as to distribute said amount of said target molecule into said at least two areas.
11. The method of claim 10 , wherein said relative motion gives rise to said amount of said target molecule being divided among at least 10 areas.
12. The method of claim 11 , wherein said relative motion gives rise to said amount of said target molecule being divided among at least 50 areas.
13. The method of claim 1 , wherein said reaction is effected at two or more areas essentially simultaneously.
14. The method of claim 1 , wherein effecting the reaction comprises heating said amount of said target molecule.
15. The method of claim 1 , wherein the introducing the sample comprising a target molecule into the device comprises introducing into an inlet in at least one of the first or second components of the device the sample comprising the target molecule.
16. The method of claim 1 , wherein effecting the reaction comprises contacting an amplification reagent with said amount of said target molecule in said at least two areas by effecting relative motion between the first and second components of the device.