SYSTEMS AND METHODS FOR ENHANCED DETECTION AND QUANTIFICATION 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 - 27 . (canceled)
28 . A kit for detecting at least one of a presence, absence, or quantity of a first and second types of target analytes in a sample comprising:
a sample collection device;
a sample analysis cartridge; and
a sample analysis reader configured to receive the sample analysis cartridge;
wherein the sample analysis cartridge comprises:
a sample preparation reservoir holding a first fluid comprising a first signaling agent to detect the first type of target analyte of the sample and a second signaling agent to detect the second type of target analyte of the sample; and a reagent ball comprising nucleic acid amplification reagents; wherein
the first type of target analyte directly or indirectly binds to the first signaling agent in the first fluid in the sample preparation reservoir, the first signaling agent having a first detectable potential for the first type of target analyte; and
the second type of target analyte directly or indirectly binds to the second signaling agent in the first fluid in the sample preparation reservoir; the second signaling agent having a second detectable potential, different from the first detectable potential, for the second type of target analyte; and
a substrate reservoir configured to hold a second fluid comprising a substrate that reacts with the first signaling agent and the second signaling agent to form a substrate-first signaling agent complex and a substrate-second signaling complex; and
an analysis channel configured to receive the second fluid from the substrate reservoir and comprising a sensor for detecting both the first and second types of target analytes based upon a first signal and a second signal respectively generated by the substrate-first signaling agent complex and the substrate-second signaling agent complex respectively following electrical stimulation of the second fluid; and
wherein the signal for each substrate-signaling agent complex is based on the respective potential of the signaling agent for the analyte.
29 . The kit of claim 28 , wherein the sample collection device comprises a distal portion adapted to be exposed to the sample, optionally wherein the sample collection device is a swab.
30 . The kit of claim 28 , wherein the sensor is an electrochemical sensor.
31 . The kit of claim 28 , wherein the sensor comprises:
a positive control working electrode configured to generate the first signal for detecting the first type of target analytes; and
a working electrode configured to generate the second signal for detecting the second type of target analytes;
wherein the positive control working electrode or the working electrode comprises a self-assembled monolayer pre-bound to an affinity molecule configured to directly or indirectly bind to the first signaling agent or the second signaling agent.
32 . The kit of claim 31 , wherein the self-assembled monolayer comprises a thiolated ethylene glycol and/or a dithiol.
33 . The kit of claim 32 , wherein the self-assembled monolayer is backfilled with a backfiller that binds directly to the affinity molecule.
34 . The kit of claim 33 , wherein the backfiller is a carboxylic acid terminated thiol.
35 . The kit of claim 34 , wherein the carboxylic acid terminated thiol is a mercaptoundecanoic acid (MUDA) or a mercaptohexanol.
36 . The kit of claim 32 , wherein the dithiol is an hexaethylene glycol dithiol (HEGT).
37 . The kit of claim 35 , wherein the self-assembled monolayer comprises an hexaethylene glycol dithiol (HEGT) and the backfiller comprises the mercaptoundecanoic acid (MUDA).
38 . The kit of claim 31 , wherein the positive control working electrode comprises the self-assembled monolayer pre-bound to affinity molecules and the working electrode comprises a self-assembled monolayer.
39 . The kit of claim 31 , further comprising a negative control working electrode configured to generate a third signal based on a reaction between the first and second signaling agents localized at the negative control working electrode and the substrate.
40 . The kit of claim 39 , wherein the negative control working electrode comprises a self-assembled monolayer.
41 . The kit of claim 40 , wherein the self-assembled monolayer of the negative control working electrode is pre-bound to affinity molecules.
42 . The kit of claim 31 , wherein the second signal is indicative of at least one of the presence, absence, or quantity of the first and second types of analytes within the sample.
43 . The kit of claim 31 , wherein the first signal is indicative of reliability of a test.
44 . The kit of claim 43 , wherein the test is determined to be reliable if the first signal indicates a quantity of the first and second types of analytes within a predetermined range.
45 . The kit of claim 39 , wherein the third signal is indicative of reliability of a test.
46 . The kit of claim 45 , wherein the test is determined to be reliable if the third signal indicates a quantity of the reaction that is below a threshold.
47 . The kit of claim 31 further comprising a processor configured to process the second signal to generate information indicative of at least one of the presence, absence, or quantity of one or more analytes within a sample.
48 . The kit of claim 28 , wherein the reagent ball comprises a plurality of forward and reverse primers, a plurality of dNTPs, a polymerase, a reverse transcriptase, a recombinase, and a single stranded binding protein.
49 . The kit of claim 48 , wherein the reagent ball further comprises a recombinase loading factor.
50 . The kit of claim 28 , wherein the reagent ball comprises nucleic acid amplification reagents for an isothermal amplification reaction.