Portable devices and methods for in situ nucleic acid detection in water samples
OwnerZhonghui Hugh Fan, Carlos Manzanas +2inventors
InventorsZhonghui Hugh Fan, Carlos Manzanas, Elise Morrison, Todd Osborne
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Abstract
The present disclosure provides for devices for preparing a sample for nucleic acid detection, methods of detection of a nucleic acid in a sample, system for preparing and analyzing nucleic acid, and the like. The present disclosure provides for an on-site testing device, system, and method for pathogen detection in water. The device, system, and method can process large sample volumes using a paper-based sample preparation method, amplify DNA using loop-mediated isothermal amplification (LAMP) or amplify RNA using reverse transcription LAMP (RT-LAMP), and detect amplicons based on color change or other signals.
Claims21
A device for preparing a sample for nucleic acid detection comprising:
- a buffer unit comprising a plurality of buffer wells arranged in a circular type shape, wherein each buffer well comprises a ball valve in a bottom of the well;
- a mixing unit comprising a mixing well and a pin, wherein the mixing unit is connected to a bottom of the buffer unit such that the mixing unit is rotatable to align with each buffer well in turn, wherein when the mixing unit is aligned with a buffer well the pin engages with the ball valve to release fluid from the buffer well into the mixing well;
- and
- a detection unit removably coupled to a bottom of the mixing unit to receive fluids from the mixing well;
- and
- a waste container coupled to the bottom of the mixing unit, wherein the waste container comprises a well that receives fluids from the detection unit.
A method of detection of a nucleic acid in a sample, comprising:
- providing a fluid sample to a first buffer well of a device, the device comprising:
- a buffer unit comprising a plurality of buffer wells arranged in a circular type shape, wherein each buffer well comprises a ball valve in a bottom of the well, wherein a first buffer well contains a lysis solution, wherein a second buffer well contains a binding buffer, wherein a third buffer contains a first wash buffer, and a fourth buffer well contains a second wash buffer;
- a mixing unit comprising a mixing well and a pin, wherein the mixing unit is connected to a bottom of the buffer unit such that the mixing unit is rotatable to align with each buffer well in turn, wherein when the mixing unit is aligned with a buffer well the pin engages with the ball valve to release fluid from the buffer well into the mixing well;
- and
- a detection unit removably coupled to a bottom of the mixing unit to receive fluids from the mixing well;
- rotating the mixing unit or the buffer unit to engage the pin with the first buffer well such that the sample and the lysis solution releases into the mixing unit;
- rotating the mixing unit or the buffer unit to engage the pin with the second buffer well such that the binding buffer releases into the mixing unit;
- rotating the mixing unit or the buffer unit to engage the pin with the third buffer well such that the first wash buffer releases into the mixing unit;
- rotating the mixing unit or the buffer unit to engage the pin with the fourth buffer well such that the second wash buffer releases into the mixing unit;
- and
- removing the detection unit once all fluids have drained from the mixing unit through the detection unit, and wherein the detection unit contains nucleic acid obtained from the fluid sample.
17-20. (canceled)
Dependencies are inferred from the claim text.