Nucleic acid isolation using polidocanol and derivatives

Abstract

This invention relates to a composition comprising a chaotropic agent, a buffering substance, and 0.5 to 5% (V/V) polidocanol or a derivative thereof. The invention is further related to uses of this composition and to a kit comprising the composition according to the invention. The invention is further related to a method for the detection of a nucleic acid in a biological sample comprising the steps of incubating the biological sample in the presence of a chaotropic agent, a buffering substance, and 0.5 to 5% (V/V) polidocanol or a derivative thereof, optionally isolating the nucleic acid, optionally amplifying the nucleic acid, and detecting the nucleic acid. The invention is further related to a method for the purification of a nucleic acid in a biological sample comprising the steps of incubating the biological sample in the presence of a chaotropic agent, a buffering substance, and 0.5 to 5% (V/V) polidocanol or a derivative thereof and isolating the nucleic acid thereby purifying the nucleic acid.

1 . A composition comprising
 a chaotropic agent,   a buffering substance, and   0.5 to 5% (V/V) polidocanol or a derivative thereof.   
     
     
         2 . The composition according to  claim 1  wherein the composition comprises 0.5 to 3% (V/V) polidocanol or a derivative thereof. 
     
     
         3 . The composition according to  claim 1  wherein the composition comprises 1 to 4.5% (V/V) polidocanol or a derivative thereof. 
     
     
         4 . The composition according to  claim 1 , wherein the chaotropic agent is selected from the group consisting of guanidinium thiocyanate, guanidinium chloride and urea. 
     
     
         5 . The composition according to  claim 1  wherein the buffering substance is selected from the group consisting Tris-(hydroxymethyl)-aminomethane (TRIS), phosphate, N-(2-hydroxyethyl)-piperazine-N′-(2-ethanesulfonic acid) (HEPES), acetate and citrate. 
     
     
         6 . The composition according to  claim 1  wherein the pH of the composition is between 3 and 5. 
     
     
         7 . The composition according to  claim 1  wherein the composition further comprises a reducing agent. 
     
     
         8 . The composition according to  claim 1  wherein the composition comprises 4 M guanidinium thiocyanate, 50 mM Na-Citrate, 1% (V/V) DTT, 3% (V/V) polidocanol, pH4. 
     
     
         9 . The composition according to  claim 1 , further comprising a protease. 
     
     
         10 . A kit comprising a composition according to  claim 1 . 
     
     
         11 . The kit according to  claim 10 , further comprising a nucleic acid binding material. 
     
     
         12 . The kit according to  claim 11 , wherein the nucleic acid binding material comprises magnetic glass particles. 
     
     
         13 . The kit according to  claim 10 , further comprising a washing buffer and an elution buffer. 
     
     
         14 . A method for the detection of a nucleic acid in a biological sample, comprising:
 a) incubating the biological sample in the presence of a chaotropic agent, a buffering substance, and 0.5 to 5% (V/V) polidocanol or a derivative thereof,   b) optionally isolating the nucleic acid,   c) optionally amplifying the nucleic acid, and   d) detecting the nucleic acid.   
     
     
         15 . The method according to  claim 14 , whereby in the amplifying step c) the nucleic acid is amplified by the polymerase chain reaction. 
     
     
         16 . A method for the purification of a nucleic acid in a biological sample, comprising:
 a) incubating the biological sample in the presence of a chaotropic agent, a buffering substance, and 0.5 to 5% (V/V) polidocanol or a derivative thereof,   b) isolating the nucleic acid, thereby purifying the nucleic acid.   
     
     
         17 . The method according to  claim 14 , wherein in step a) the biological sample is incubated in the presence of a chaotropic agent, a buffering substance, and 0.5 to 3% (V/V) polidocanol or a derivative thereof. 
     
     
         18 . The method according to  claim 14 , whereby the isolating step b) comprises binding the nucleic acid to a nucleic acid binding material, optionally washing the nucleic acid bound to the material and eluting the nucleic acid from the material. 
     
     
         19 . The method according to  claim 18 , wherein the nucleic acid binding material comprises magnetic glass particles. 
     
     
         20 . The method according to  claim 14 , wherein the biological sample is a fluid from the human or animal body, said fluid being selected from the group consisting of blood, blood plasma, blood serum and urine. 
     
     
         21 . The method according to  claim 14 , wherein the nucleic acid comprises DNA or RNA or both. 
     
     
         22 . The method according to  claim 14 , wherein the nucleic acid is derived from a microorganism or a virus. 
     
     
         23 . The method according to  claim 14 , wherein the nucleic acid is derived from a member of the group consisting of a hepatitis A virus, hepatitis B virus, the hepatitis C virus, the human immunodeficiency virus or the cytomegalovirus.