System and method for rapid multiplexed sample processing with applications for nucleic acid amplification assays
The invention(s) cover systems and methods for target detection in a multiplexed and rapid manner. Embodiments of the system can include: a base substrate; and an array of sample processing regions defined at a broad surface of the base substrate, wherein each of the array of sample processing regions includes: a set of microwell subarrays arranged in a gradient by volumetric capacity between an upstream end and a downstream end of each respective sample processing region, and a boundary separating each respective sample processing region from adjacent sample processing regions. The system can support methods, with example implementation by an automated platform, for returning preliminary results from a subset of microwells of the samples processing regions, as well as results pertaining to specific and non-specific amplification, for multiple targets of a sample.
1. A system for target detection, the system comprising:
a base substrate shaped as a microwell plate;
an array of sample processing regions comprising at least three sample processing regions defined at a top surface of the base substrate, wherein a sample processing region of the array of sample processing regions comprises:
a set of microwell subarrays arranged to provide up to a 9 log dynamic range for a polymerase chain reaction (PCR) process, based upon distribution of the set of microwell subarrays in a gradient by volumetric capacity between a first end and a second end of the sample processing region, the set of microwell subarrays comprising:
an initial subarray with microwells individually having a first volumetric capacity and first cross section smaller than microwells of all other subarrays of the set of microwell subarrays, and
a terminal subarray with microwells individually having a second volumetric capacity and second cross section greater than microwells of all other subarrays of the set of microwell subarrays, wherein the first volumetric capacity and the second volumetric capacity each fall within a range from 0.1 nL to 100 nL, and
a boundary recessed into the base substrate, surrounding the sample processing region as a moat, and separating the sample processing region from adjacent sample processing regions of the array of sample processing regions;
a heating subsystem comprising a heating body and a temperature sensor;
a cooling subsystem; and
a controller coupled to the heating subsystem and the cooling subsystem for performance of the PCR process.
2. The system of claim 1 , wherein the base substrate is configured to absorb inhibitors of target detection reactions within the array of sample processing regions, wherein said inhibitors comprise RNases, DNases, and proteinases.
3. The system of claim 1 , wherein the array of sample processing regions comprises from 3 to 96 sample processing regions.
4. The system of claim 1 , wherein each of the set of microwell subarrays comprises from 10 through 500,000 microwells.
5. The system of claim 1 , wherein the initial microwell subarray occupies a first footprint of the base substrate, and wherein the terminal microwell subarray occupies a second footprint of the base substrate larger than the first footprint.
6. The system of claim 1 , wherein the boundary comprises a recessed channel operable to receive sample overflow.
7. The system of claim 1 , further comprising a cover layer biased against the base substrate, the cover layer sealing individual sample processing regions of the array of sample processing regions.
8. The system of claim 7 , wherein a surface of the cover layer facing the array of sample processing regions of the base substrate has a surface roughness less than 5 microns and is configured to reflect light toward interior volumes of the array of sample processing regions.
9. The system of claim 7 , wherein the cover layer comprises a photo-thermal coating configured to receive incident light of a specified wavelength range, transform incident light into heat, and transmit said heat to contents of the array of sample processing regions.
10. The system of claim 7 , wherein the cover layer comprises a first locking portion complementary to a second locking portion of the base substrate, wherein the cover layer is composed of an elastomeric material, and wherein mating of the cover layer to the base substrate spreads a sample across the sample processing region.
11. The system of claim 7 , wherein the cover layer seals individual sample processing regions of the array of sample processing regions, and transfers heat to contents of the array of sample processing regions upon receiving heat from a heating subsystem.
12. The system of claim 7 , wherein the cover layer is separated from the base substrate by a film layer.