Rapid diagnostics
Systems and methods for rapid diagnostics related to the use of isothermal amplification reagents for detection of microbial species, including coronavirus, and methods of use, are provided.
1. A cartridge comprising at least a first and second ampoule, a lysis chamber, an amplification chamber, and a sample receiving chamber, wherein the first ampoule comprises DNA isolation solution capable of isolating a target RNA from a crude or unprocessed sample comprising a cell or virus containing the target RNA and the second ampoule comprises a loop-mediated isothermal amplification (LAMP) composition for detecting the presence of the target RNA in a sample, comprising optimized LAMP primers consisting of SEQ ID NOs: 61983-61988 and amplification reagents for single amplification and glycine or taurine, or a combination thereof for amplifying the target RNA in less than one hour.
2. The cartridge of claim 1 , configured to fit in a system comprising a heating means, an optic means, a means for releasing reagents on the cartridge, and a means for readout of assay result.
3. The cartridge of claim 1 , wherein the contents of the first ampoule and/or second ampoule are lyophilized.
4. A device designed to receive one or more cartridges of claim 2 and further comprising one or more motors connected to a plunger for rupturing of the first and second ampoule of the cartridge and configured within the device to align with the first and second ampoule of the inserted cartridge, a heating element configured to align with the amplification chamber of the inserted cartridge, an optical detector configured to align with the amplification chamber of the inserted cartridge, and a display.
5. The device of claim 4 , further comprising a graphical user interface for programming the device and/or readout of the results of the assay.
6. A system comprising a docking station and two or more devices of claim 5 , wherein the docking station is configured to receive the two or more devices.
7. The cartridge of claim 1 , wherein the second ampoule comprises isothermal amplification reagents and a CRISPR-Cas collateral detection system for amplifying and detecting a target RNA.
8. The cartridge of claim 7 , wherein the target RNA is a coronavirus.
9. The cartridge of claim 1 , wherein the DNA isolation solution and/or the lysis well comprises RNA binding beads.
10. The cartridge of claim 1 , wherein the DNA isolation solution comprises a protease.
11. The cartridge of claim 10 , wherein the protease is proteinase K.
12. The cartridge of claim 1 , wherein the amplification reagents comprise reverse transcriptase.
13. The cartridge of claim 1 , wherein the first ampoule comprises isolation solution in an amount to provide a ratio from 2:1 to 1:2 isolation solution:crude or unprocessed sample.
14. The cartridge of claim 1 , wherein the LAMP composition further comprises colorimetric or fluorescent detection reagents.