Compositions and methods for rapid covid-19 detection
AssigneeDuke University
InventorsXin Song, John Reif
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Abstract
The present disclosure provides compositions and methods related to the detection of pathogenic organisms. In particular, the present disclosure provides compositions and methods related to the detection and/or quantification of viral RNA in a sample from a subject that has, or is suspected of having, a SARS-COV-2 infection. Using rapid reverse-transcription loop-mediated isothermal amplification (RT-LAMP), the compositions and methods of the present disclosure provide a portable, inexpensive, rapid, and accurate assay platform for detecting and/or quantifying the presence of a pathogenic organism (e.g., SARS-COV-2) in a patient sample.
Claims41
A composition for performing a reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction, the composition comprising:
- a reaction buffer comprising a DNA polymerase and a reverse transcriptase;
- a chaotropic agent;
- and
- at least one excipient;
- wherein the composition is lyophilized to form an RT-LAMP reaction mixture.
A method for generating a reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction mixture, the method comprising:
- combining a reaction buffer comprising a DNA polymerase and a reverse transcriptase with a chaotropic agent and at least one excipient into a container;
- and
- subjecting the container to a lyophilization process to form an RT-LAMP reaction mixture.
The method of claim 29, wherein the chaotropic agent is selected from the group consisting of n-butanol, ethanol, guanidine hydrochloride, guanidine thiocyanate, lithium perchlorate, lithium acetate, magnesium chloride, phenol, 2-propanol, sodium dodecyl sulfate, sodium iodide, thiourea, and urea.
The method of any of claims 29 to 31, wherein the guanidine hydrochloride is present at a concentration prior to lyophilization sufficient to produce a reconstituted concentration ranging from about 20 mM to about 80 mM.
Dependencies are inferred from the claim text.