Nucleic acid preservation solution and methods of manufacture and use

Abstract

Disclosed is nucleic acid preserving compositions and methods of manufacturing and using the same. Compositions include a carrier, a chaotropic agent, a buffering agent, a chelating agent, a surfactant, an alcohol, an acid, and a mucolytic agent. Compositions as aqueous solutions can include water as a carrier. Preferred embodiments include water, guanidine thiocyanate, Tris, EDTA, SLS, SDA 3C, HCl, and N-acetyl-L-cysteine. Some embodiments include a colored dye as a visual indicator. Methods of manufacturing include combining the components into a mixture, such as an aqueous solution. Methods of use include providing a biological sample that includes nucleic acid and contacting the biological sample with the composition. Kits include the composition disposed in a portion of a biological sample collection apparatus.

We claim: 
     
         1 . A nucleic acid preservation composition, comprising:
 an aqueous carrier;   about 20% to about 50%, w/w, of a chaotropic agent;   about 0.1% to about 5%, w/w, of a buffering agent;   about 0.05% to about 2.5%, w/w, of a chelating agent comprising;   about 0.01% to about 5%, w/w, of a detergent or a surfactant;   about 5% to about 25%, w/w, of an alcohol, wherein the alcohol comprises ethanol and optionally comprises isopropanol; and   0.005% to about 0.5%, w/w, of a mucolytic agent or a reducing agent, wherein the mucolytic agent or reducing agent is other than dithiothreitol and/or 2-mercaptoethanol,   the composition having a pH of 6.5-9.5.   
     
     
         2 . The composition of  claim 1 , wherein the composition has a pH of about 7 to about 9. 
     
     
         3 . The composition of  claim 1 , wherein:
 the chaotropic agent comprises guanidine thiocyanate;   the buffering agent comprises tris(hydroxymethyl)aminomethane (Tris);   the chelating agent comprises ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate; and   the detergent or surfactant comprises sodium lauroyl sarcosinate (SLS).   
     
     
         4 . The composition of  claim 1 , wherein the composition is substantially free or devoid of antimicrobial agent(s), bactericidal agent(s), and/or bacteriostatic agent(s) other than the chaotropic agent, the detergent or surfactant, the alcohol, and the mucolytic agent or reducing agent. 
     
     
         5 . The composition of  claim 1 , wherein the composition is substantially free or devoid of protease(s). 
     
     
         6 . The composition of  claim 1 , wherein the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the chaotropic agent, the surfactant, the alcohol, and the mucolytic agent. 
     
     
         7 . A nucleic acid preservation kit, comprising:
 a sample collection apparatus comprising a solution compartment; and   the nucleic acid preservation composition of  claim 1  disposed in the solution compartment.   
     
     
         8 . A method of preserving nucleic acid in a biological sample, the method comprising:
 obtaining or providing a biological sample containing nucleic acid, the biological sample optionally comprising saliva; and   contacting the biological sample with the nucleic acid preservation composition of  claim 1 .   
     
     
         9 . A nucleic acid preservation composition, comprising:
 guanidine thiocyanate;   tris(hydroxymethyl)aminomethane (Tris);   ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate;   sodium lauroyl sarcosinate (SLS);   an alcohol, wherein the alcohol comprises ethanol and optionally comprises isopropanol; and   a mucolytic agent or reducing agent other than dithiothreitol and/or 2-mercaptoethanol.   
     
     
         10 . The composition of  claim 9 , wherein the composition has a pH of 6.5-9.5. 
     
     
         11 . The composition of  claim 9 , wherein the composition has a pH of 7-9. 
     
     
         12 . The composition of  claim 9 , wherein the composition is substantially free or devoid of antimicrobial agent(s), bactericidal agent(s), and/or bacteriostatic agent(s) other than the chaotropic agent, the detergent or surfactant, the alcohol, and the mucolytic agent or reducing agent. 
     
     
         13 . The composition of  claim 9 , wherein the composition is substantially free or devoid of protease(s). 
     
     
         14 . The composition of  claim 9 , wherein the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the chaotropic agent, the surfactant, the alcohol, and the mucolytic agent. 
     
     
         15 . A nucleic acid preservation kit, comprising:
 a sample collection apparatus comprising a solution compartment; and   the nucleic acid preservation composition of  claim 9  disposed in the solution compartment.   
     
     
         16 . A method of preserving nucleic acid in a biological sample, the method comprising:
 obtaining or providing a biological sample containing nucleic acid, the biological sample optionally comprising saliva; and   contacting the biological sample with the nucleic acid preservation composition of claim.   
     
     
         17 . A nucleic acid preservation composition, comprising:
 an aqueous carrier;   a chaotropic agent;   a buffering agent;   a chelating agent;   a detergent or a surfactant;   an alcohol, wherein the alcohol comprises ethanol and optionally comprises isopropanol; and   a mucolytic agent or reducing agent other than dithiothreitol and/or 2-mercaptoethanol.   
     
     
         18 . The nucleic acid preservation composition of  claim 17 , wherein the composition has a pH of about 7 to about 9. 
     
     
         19 . The nucleic acid preservation composition of  claim 18 , wherein:
 the composition is substantially free or devoid of antimicrobial agent(s), bactericidal agent(s), and/or bacteriostatic agent(s) other than the chaotropic agent, the detergent or surfactant, the alcohol, and the mucolytic agent or reducing agent;   the composition is substantially free or devoid of protease(s); and   the composition is substantially free or devoid of inhibitor(s) of ribonuclease other than the chaotropic agent, the surfactant, the alcohol, and the mucolytic agent.   
     
     
         20 . The nucleic acid preservation composition of  claim 19 , wherein:
 the chaotropic agent comprises guanidine thiocyanate;   the buffering agent comprises tris(hydroxymethyl)aminomethane (Tris);   the chelating agent comprises ethylenediaminetetraacetic acid (EDTA) or EDTA disodium (salt) dihydrate; and   the detergent or surfactant comprises sodium lauroyl sarcosinate (SLS).