Compositions for stabilizing DNA, RNA and proteins in saliva and other biological samples during shipping and storage at ambient temperatures
First Claim
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1. An admixture comprising:
- A) a composition for substantially stable storage of nucleic acid and polypeptide molecules in a biological sample at room temperature, comprising;
(a) a compound selected from the group consisting of;
1-methyl-3-carboxyethyl-imidazolium bromide,1-hexyl-3-methylimidazolium bromide,1-octyl-3-methylimidazolium bromide,1-decyl-3-methylimidazolium bromide, and1-(2-hydroxyethyl)-3-methylimidazolium bromide;
(b) one, two, or all three of;
(i) at least one precipitating agent,(ii) at least one lower alcohol, and(iii) at least one chaotrope;
(c) one or more of;
(i) a chelating agent,(ii) a reducing agent,(iii) a pH buffer, and(iv) water; and
(d) a surfactant or detergent,wherein the composition is an aqueous solution and the compound of (a) is present in the composition at a concentration of about 0.1% to about 10% (w/v); and
B) a biological sample comprising saliva or sputum.
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Abstract
Compositions and methods are disclosed for substantially liquid, gel, suspension, slurry, semisolid and/or colloid storage of biological samples following admixture with the herein disclosed storage composition, permitting substantial recovery of biological activity following storage without refrigeration. In certain embodiments, unfractionated saliva samples may be stored without refrigeration for weeks, months or years in a form that permits recovery of intact DNA following the storage period.
331 Citations
24 Claims
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1. An admixture comprising:
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A) a composition for substantially stable storage of nucleic acid and polypeptide molecules in a biological sample at room temperature, comprising; (a) a compound selected from the group consisting of; 1-methyl-3-carboxyethyl-imidazolium bromide, 1-hexyl-3-methylimidazolium bromide, 1-octyl-3-methylimidazolium bromide, 1-decyl-3-methylimidazolium bromide, and 1-(2-hydroxyethyl)-3-methylimidazolium bromide; (b) one, two, or all three of; (i) at least one precipitating agent, (ii) at least one lower alcohol, and (iii) at least one chaotrope; (c) one or more of; (i) a chelating agent, (ii) a reducing agent, (iii) a pH buffer, and (iv) water; and (d) a surfactant or detergent, wherein the composition is an aqueous solution and the compound of (a) is present in the composition at a concentration of about 0.1% to about 10% (w/v); and B) a biological sample comprising saliva or sputum. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method for substantially stabilizing one or a plurality of nucleic acid or polypeptide molecules that are present in a biological sample comprising saliva or sputum, comprising:
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(a) admixing the biological sample comprising saliva or sputum with a composition comprising (A) a compound selected from the group consisting of; 1-methyl-3-carboxyethyl-imidazolium bromide, 1-hexyl-3-methylimidazolium bromide, 1-octyl-3-methylimidazolium bromide, 1-decyl-3-methylimidazolium bromide, and 1-(2-hydroxyethyl)-3-methylimidazolium bromide; (B) one, two, or all three of; (i) at least one precipitating agent, (ii) at least one lower alcohol, and (iii) at least one chaotrope; (C) one or more of; (i) a chelating agent, (ii) a reducing agent, (iii) a pH buffer, and (iv) water; and (D) a surfactant or detergent, wherein the composition is an aqueous solution and the compound of (A) is present in the composition at a concentration of about 0.1% to about 10% (w/v), to obtain a mixture; and (b) maintaining the mixture without refrigeration for a time period of at least 7 days, and thereby substantially stabilizing said one or a plurality of nucleic acid or polypeptide molecules that are present in the biological sample, wherein either one, two or all three of; (1) degradation is substantially prevented of at least 70% of recoverable DNA in the mixture, relative to an amount of DNA or polypeptide that is recoverable from the sample when stored for the time period at −
20°
C. without the composition,(2) degradation is substantially prevented of at least 70% of recoverable RNA in the mixture, relative to an amount of RNA that is recoverable from the sample when stored for the time period at −
80°
C. without the composition, and(3) degradation is substantially prevented of at least 70% of recoverable polypeptide molecules in the mixture, relative to an amount of polypeptide molecules that is recoverable from the sample when stored for the time period at −
20°
C. without the composition. - View Dependent Claims (20, 21, 22, 23, 24)
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Specification