Cartridges, kits, and methods for amplification and detection of analytes
Devices, systems, and methods for detecting molecules of interest within a collected sample are described herein. In certain embodiments, self-contained sample analysis systems are disclosed, which include a reusable reader component, a disposable cartridge component, and a disposable sample collection component. The reader component may communicate with a remote computing device for the digital transmission of test protocols and test results. In various disclosed embodiments, the systems, components, and methods are configured to identify the presence, absence, and/or quantity of particular nucleic acids, proteins, or other analytes of interest, for example, in order to test for the presence of one or more pathogens or contaminants in a sample.
1. A method of amplification for determining at least one of a presence, absence, or quantity of a target nucleic acid in a sample, the method comprising:
introducing the sample comprising the target nucleic acid into fluid held by a reservoir of a microfluidic cartridge and introducing a reagent ball into the fluid held by the reservoir, the reagent ball comprising reagents for amplification of the target nucleic acid by an amplification reaction;
amplifying the target nucleic acid with an effective amount of the reagents for amplification in the reservoir to generate amplicons comprising signaling agents;
introducing the fluid comprising the amplicons into an analysis channel from the reservoir such that the amplicons indicative of at least one of a presence, absence, or quantity of the target nucleic acid in the sample localize over a sensor at least partially disposed in the analysis channel;
reacting the signaling agents with a substrate; and
sensing signals indicative of at least one of the presence, absence, or quantity of the target nucleic acid in the sample based on reactions between the substrate and the signaling agents of the amplicons localized over the sensor.
2. The method of claim 1 , wherein the reagent ball comprises polymerase, primers for amplification of the target nucleic acid and the signaling agent for the detection of the amplification of the target nucleic acid.
3. The method of claim 1 , wherein the reagent ball further comprises one or more affinity molecules covalently or non-covalently bound to a solid particle for detection of the target nucleic acid.
4. The method of claim 1 , wherein introducing the reagent ball comprises moving the reagent ball from a pre-mixing position, wherein the reagent ball is disposed in the input tunnel and not exposed to the fluid in the reservoir, to a mixing position, wherein the reagent ball is exposed to the fluid in the reservoir.
5. The method of claim 1 , wherein introducing the sample into the fluid held by the reservoir comprises moving a shuttle having the sample disposed therein into the reservoir.
6. The method of claim 5 , wherein introducing the reagent ball into the fluid held by the reservoir comprises moving the shuttle having the reagent ball disposed therein into the reservoir.
7. The method of claim 5 , further comprising moving the fluid, the sample, and the reagents between first and second compartments of the shuttle to mix the fluid, the sample, and the reagents in the reservoir.
8. The method of claim 7 , wherein moving the fluid, the sample, and the reagents comprises emitting energy from a piezoelectric transducer into the reservoir to move the fluid in the reservoir between the first and second compartments.
9. The method of claim 1 , further comprising sensing temperature indicative of temperature of the fluid in the reservoir.
10. The method of claim 9 , further comprising emitting energy into the reservoir to move the fluid in the reservoir to mix the fluid, the sample, and the reagents in the reservoir; and
further emitting energy in a modified manner based on the sensed temperature.
11. The method of claim 10 , wherein emitting the energy causes the amplification reaction to cure on the target nucleic acid.
12. The method of claim 1 , further comprising sensing insertion of a sample collection device in the input tunnel.
13. The method of claim 1 , further comprising introducing a substrate solution comprising the substrate from a substrate reservoir into the analysis channel to the sensor such that the substrate reacts with the signaling agents localized over the sensor.
14. The method of claim 13 , further comprising, before introducing the substrate solution, introducing a wash solution from a wash reservoir into the analysis channel to the sensor.