Rapid diagnostic test for LAMP
Kits and methods are described that are directed to specific and sensitive methods of target nucleic acid detection and more specifically detecting target nucleic acids directly from biological samples. The kits and methods were developed to be easy to use involving a minimum number of steps and giving rapid and consistent results either at point of care or in high throughput situations. The kits and methods utilize in various combinations, reversible inhibitors of kit components, thermolabile enzymes, poloxamers, various salts, indicators and one or more Loop-Mediated Isothermal Amplification (LAMP) primer sets for detecting single and/or multiple targets and variants of the targets including SARS-CoV-2 targets and variants thereof in a single reaction. The kits and methods permit detection of the target nucleic with similar sensitivity regardless of the presence of undefined mutations that may enhance the virulence of cells or viruses containing the undefined mutations.
1. A facemask for detecting a pathogen, the face mask comprising:
(a) A lysis reagent for disrupting the pathogen received by the mask in an aqueous aerosol, to release nucleic acid;
(b) a lyophilized isothermal amplification mixture; the lyophilized amplification mixture comprising a polymerase, and one or more primers for hybridizing to the nucleic acid; and
(c) a signaling reagent for reporting a positive amplification reaction.
2. The facemask according to claim 1 , wherein the aqueous aerosol is saliva or nasal effluent from a person other than the mask wearer.
3. The facemask according to claim 1 , wherein the lysis mixture is compartmentally separated from the amplification mixture.
4. The facemask according to claim 1 , wherein the facemask further comprises a reagent in the amplification mixture capable of generating an exothermic chemical reaction when contacted by the lysis mixture after contact with the aqueous aerosol, wherein the exothermic reaction initiates the amplification reaction.
5. The facemask according to claim 1 , wherein the signaling reagent produces a color change caused by release of pyrophosphate.
6. The facemask according to claim 1 , wherein the signaling reagent is a sound.
7. The facemask according to claim 1 , wherein the lyophilized amplification mixture is immobilized on a substrate.
8. The facemask according to claim 7 , wherein the immobilized substrate is beads.
9. A method for detecting a pathogen in an environment, comprising:
(a) obtaining a fabric containing a lysis reagent, lyophilized isothermal amplification mixture; and a signaling reagent;
(b) receiving an aerosol from a third party or from the wearer of the fabric;
(c) lysing the pathogen to release nucleic acids by reagents in the fabric;
(d) amplifying the nucleic acids by amplification reagents in the fabric; and
(e) producing a visible or audible signal if the nucleic acids from the pathogen are present in the received aerosol.