FTA-coated media for use as a molecular diagnostic tool
There is provided a substrate for lysing cells and purifying nucleic acid having a matrix and a coating and an integrity maintainer for maintaining the purified nucleic acid. Also provided is a method of purifying nucleic acid by applying a nucleic acid sample to a substrate having an anionic detergent affixed to a matrix, the substrate physically capturing the nucleic acid, bonding the nucleic acid to a substrate and generating a signal when the nucleic acid bonds to the substrate indicating the presence of the nucleic acid. A kit for purifying nucleic acid containing a coated matrix and an integrity maintenance provider for preserving the matrix and purifying nucleic acid is also provided.
1. A kit for indicating the presence of nucleic acid in a sample, the kit comprising:
a. a dry substrate for lysing cells and purifying nucleic acid therefrom, said dry substrate consisting of:
i. a solid matrix, wherein the solid matrix comprises nitrocellulose or nylon; and
ii. a coating sorbed to the solid matrix, wherein the coating comprises a cellular lysis reagent comprising an anionic surfactant or detergent at a concentration of at least 5% which induces cellular lysis and releases or stabilizes the nucleic acid onto the dry substrate, and
b. an indicator which detects the presence of nucleic acid, wherein the nucleic acid is maintained on the solid matrix, the indicator comprising an external substance which generates a signal and detects the presence of the nucleic acid immobilized thereon.
2. The kit according to claim 1 , wherein said indicator is selected from the group consisting essentially of a fluorescent indicator, color indicator or photometric indicator.
3. The kit according to claim 1 , wherein said substrate is in a shape selected from the group consisting essentially of a swab, a sheet, a card, and a ball.
4. The kit according to claim 3 , wherein said substrate further includes an integrity maintenance means which reduces degradation or loss of the matrix or the nucleic acid.
5. The kit according to claim 4 , wherein when said substrate is a sheet, said integrity maintenance means is a plastic bag.
6. A kit for purifying nucleic acid comprising:
a. a dry substrate comprising:
i. a solid matrix, wherein the solid matrix comprises nitrocellulose or nylon; and
ii. a coating sorbed to the solid matrix, wherein the coating comprises a cellular lysis reagent comprising an anionic surfactant or detergent at a concentration of at least 5% which induces cellular lysis and releases or stabilizes the nucleic acid onto the dry substrate, and
b. an indicator which detects the presence of nucleic acid, wherein the nucleic acid is maintained on the solid matrix, the indicator comprising an external substance which generates a signal and detects the presence of the nucleic acid immobilized thereon; and
c. an integrity maintenance means which reduces degradation or loss of the matrix or the nucleic acid.
7. The kit according to claim 6 , wherein said coated matrix is in a shape selected from the group consisting essentially of a swab, a sheet, a card, and a ball.
8. The kit according to claim 6 , wherein said coated matrix is in a shape selected from the group consisting essentially of a plastic bag, cellophane, a sealable container, cartridge and parafilm.
9. A blood card for labeling blood transfusion bags comprising:
a. a dry substrate comprising a solid matrix selected from the group consisting of nitrocellulose, carboxymethylcellulose, polyester, polyamide, polytetrafluoroethylene and porous ceramics, wherein the solid matrix further comprises a chemical coating sorbed to the solid matrix, the chemical coating comprising:
i. a weak base;
ii. a chelating agent; and
iii. a cellular lysis reagent comprising an anionic surfactant or detergent at a concentration of at least 5% which induces cellular lysis and releases or stabilizes the nucleic acid onto the dry substrate; and
b. an indicator which detects the presence of nucleic acid, wherein the nucleic acid is maintained on the solid matrix, the indicator comprising an external substance which generates a signal and detects the presence of the nucleic acid immobilized thereon; and
c. an integrity maintenance means which reduces degradation or loss of the matrix or the nucleic acid.
10. The kit of claim 1 , wherein the anionic surfactant or detergent is selected from the group consisting of sodium dodecyl sulfate (SDS), alky aryl sulfonates, sodium tetradecylsulfate long chain (fatty) alcohol sulfates, sodium 2-ethyihexysulfate olefine sulfates, sulfosuccinates or phosphate esters.
11. The kit of claim 10 , wherein:
a. the weak base comprises a Tris or trishydroxymethyl methane; and
b. the chelating agent comprises ethylenediamine tetra-acetic acid (EDTA).
12. The kit of claim 1 , wherein the anionic surfactant or detergent comprises sodium dodecyl sulfate at a concentration of at least 5% to about 10%.
13. The kit of claim 1 , wherein the coating further comprises a free radical trap.
14. The kit of claim 6 , wherein the anionic surfactant or detergent is selected from the group consisting of sodium dodecyl sulfate (SDS), alky aryl sulfonates, sodium tetradecylsulfate long chain (fatty) alcohol sulfates, sodium 2-ethylhexysulfate olefine sulfates, sulfosuccinates or phosphate esters.
15. The kit of claim 14 , wherein:
a. the weak base comprises a Tris or trishydroxymethyl methane; and
b. the chelating agent comprises ethylenediamine tetra-acetic acid (EDTA).
16. The kit of claim 6 , wherein the anionic surfactant or detergent comprises sodium dodecyl sulfate at a concentration of at least 5% to about 10%.
17. The kit of claim 6 , wherein the chemical coating further comprises a free radical trap.
18. The kit of claim 6 , wherein the indicator is selected from the group consisting essentially of a fluorescent indicator, color indicator or photometric indicator.
19. A kit for labeling blood transfusion bags comprising:
a. a dry substrate comprising a solid matrix selected from the group consisting of nitrocellulose, carboxymethylcellulose, polyester, polyamide, polytetrafluoroethylene and porous ceramics, the solid matrix being coated with a chemical coating sorbed to the solid matrix, the chemical coating comprising a weak base, a chelating agent, and a cellular lysis reagent comprising an anionic surfactant or detergent at a concentration of at least 5% which induces cellular lysis and releases or stabilizes the nucleic acid onto the dry substrate;
b. an indicator which detects the presence of nucleic acid, wherein the nucleic acid is maintained on the solid matrix, the indicator comprising an external substance which generates a signal and detects the presence of the nucleic acid immobilized thereon; and
c. an integrity maintenance means which reduces degradation or loss of the matrix or the nucleic acid.
20. A kit for purifying nucleic acid comprising:
a. a dry substrate for lysing cells and purifying nucleic acid comprising a solid matrix comprising nitrocellulose, the solid matrix being coated with a chemical coating sorbed to the solid matrix, the chemical coating comprising:
i. a weak base;
ii. a chelating agent; and
iii. a cellular lysis reagent comprising an anionic surfactant or detergent at a concentration of at least 5% which induces cellular lysis and releases or stabilizes the nucleic acid onto the dry substrate; and
b. an indicator which detects the presence of nucleic acid, wherein:
i. the indicator comprises an external substance which generates a signal and detects the presence of the nucleic acid immobilized thereon; and
ii. the indicator comprises a polyethyleneimine conjugate or an enzyme-linked immunosorbant assay (ELISA) reagent.
21. The kit of claim 20 , wherein the anionic surfactant or detergent is selected from the group consisting of sodium dodecyl sulfate (SDS), alky aryl sulfonates, sodium tetradecylsulfate long chain (fatty) alcohol sulfates, sodium 2-ethylhexysulfate olefine sulfates, sulfosuccinates or phosphate esters.
22. The kit of claim 20 , wherein:
a. the weak base comprises a Tris or trishydroxymethyl methane; and
b. the chelating agent comprises ethylenediamine tetra-acetic acid (EDTA).
23. The kit of claim 20 , wherein the anionic surfactant or detergent comprises sodium dodecyl sulfate at a concentration of at least 5% to about 10%.
24. The kit of claim 20 , wherein the chemical coating further comprises a free radical trap.
25. The kit of claim 20 , wherein the nucleic acid comprises a sequence of interest and the indicator comprises an enzyme-linked immunosorbant assay (ELISA) using antibodies to the sequence of interest.
26. The kit of claim 1 , wherein the indicator comprises a polyethyleneimine conjugate or an enzyme-linked immunosorbant assay (ELISA) reagent.
27. The kit of claim 4 , wherein the integrity maintenance means reduces contact of the matrix of the dry substrate, or nucleic acid stored thereon, with a contaminant.
28. The kit of claim 6 , wherein the integrity maintenance means reduces contact of the matrix of the dry substrate, or nucleic acid stored thereon, with a contaminant.
29. The kit of claim 9 , wherein the integrity maintenance means reduces contact of the matrix of the dry substrate, or nucleic acid stored thereon, with a contaminant.
30. The kit of claim 19 , wherein the integrity maintenance means reduces contact of the matrix of the dry substrate, or nucleic acid stored thereon, with a contaminant.
31. The kit of claim 20 , wherein the integrity maintenance means reduces contact of the matrix of the dry substrate, or nucleic acid stored thereon, with a contaminant.