Vessel for blood sampling

Abstract

The present invention relates to a vessel for withdrawing blood, the vessel containing a solution which comprises a guanidinium salt, a buffer substance, a reducing agent, and/or a detergent as components. The vessel is particularly suited for withdrawing blood which is to be analyzed with respect to nucleic acids.

1 - 160 . (canceled) 
     
     
         161 . A blood withdrawing vessel comprising
 an evacuated chamber which is provided for receiving withdrawn blood containing a nucleic acid-stabilizing aqueous solution for stabilizing nucleic acids for quantitative analysis in withdrawn blood directly upon contact with the solution,   wherein the solution comprises the following components:   a guanidinium salt in a concentration of 1 to 8.0 M;   a buffer substance in a concentration of 10 to 300 mM; and   a detergent.   
     
     
         162 . The vessel according to  claim 161 , characterized in that the guanidinium salt is selected from guanidinium thiocyanate and guanidinium chloride. 
     
     
         163 . The vessel according to  claim 161 , characterized in that the guanidinium salt is present in a concentration of 2.5 to 8.0 M. 
     
     
         164 . The vessel according to  claim 161 , characterized in that the buffer substance is selected from Tris, HEPES, MOPS, citrate and phosphate buffer. 
     
     
         165 . The vessel according to  claim 161 , characterized in that the detergent is selected from Triton-X-100, NP-40, polydocanol and Tween 20. 
     
     
         166 . The vessel according to  claim 161 , characterized in that the pH of the solution is between 4.0 and 7.5. 
     
     
         167 . The vessel according to  claim 161 , characterized in that it contains withdrawn blood. 
     
     
         168 . A method of withdrawing blood, comprising the step of directly introducing the blood into a vessel according to  claim 161 . 
     
     
         169 . The method according to  claim 168 , characterized in that an amount of blood is withdrawn that is 0.1 to 4 times the volume of the solution in the vessel. 
     
     
         170 . The method according to  claim 169 , characterized in that the concentration of the guanidinium salt after the blood is introduced is between 1.0 M and 5 M. 
     
     
         171 . A method for quantitatively stabilizing nucleic acids from blood, comprising the step of introducing blood into a vessel according to  claim 161  and, optionally, isolating the nucleic acids with conventional methods. 
     
     
         172 . The method according to  claim 170 , characterized in that the pH of the resultant mixture of the solution and blood is between 5.0 and 7.6. 
     
     
         173 . The method according to  claim 172 , characterized in that the concentration of the guanidinium salt, after blood is introduced, is between 1.5 and 5 M. 
     
     
         174 . A vessel comprising an interior, the interior comprising an aqueous solution that comprises a guanidinium salt, a buffer and a detergent, wherein the interior is evacuated, and wherein the aqueous solution is capable of lysing blood cells and stabilizing nucleic acids for quantitative analysis. 
     
     
         175 . The vessel of  claim 174 , wherein the vessel comprises a tube having an open end sealed by a septum. 
     
     
         176 . The vessel of  claim 174 , wherein the evacuation is effective for drawing a specific volume of a fluid sample into the interior. 
     
     
         177 . The vessel of  claim 174 , wherein the guanidinium salt, the buffer and the detergent are present in amounts effective to provide cell lysis and stabilization of nucleic acids in a fluid sample by inactivation of nucleases. 
     
     
         178 . The vessel of  claim 177 , wherein the buffer is effective, upon mixing with the specific volume of a blood sample, to provide the resultant mixture with a pH of 5.0 to 7.6. 
     
     
         179 . The vessel of  claim 178 , wherein the buffer is effective to provide the resultant mixture with a pH of 6.3 to 6.9. 
     
     
         180 . The vessel of  claim 174 , wherein the pH of the aqueous solution in the vessel is 4.0 to 7.5. 
     
     
         181 . The vessel of  claim 180 , wherein the pH is 5 to 6. 
     
     
         182 . The vessel of  claim 176 , wherein the specific volume is about 0.1 to about 4 times the volume of the solution. 
     
     
         183 . The vessel of  claim 174 , wherein the guanidinium salt is one or more salts selected from the group consisting of guanidinium thiocyanate and guanidinium chloride. 
     
     
         184 . The vessel of  claim 174 , wherein the buffer is selected from the group consisting of: Tris (tris(hydroxymethyl)aminomethane), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), MOPS (4-morpholinepropanesulfonic acid), MES (4-morpholineethanesufonic acid), citrate and phosphate buffer. 
     
     
         185 . The vessel of  claim 174 , wherein the detergent is selected from the group consisting of: (polyethylene glycol tert-octylphenyl ether), (polyethylene glycol 4-nonylphenyl ether), Polydocanol (dodecyl polyethylene glycol ether) and (polyethylene glycol sorbitan monolaurate). 
     
     
         186 . The vessel of  claim 174 , wherein the solution comprises a single buffer and a single detergent. 
     
     
         187 . The vessel of  claim 174 , wherein the aqueous solution consists essentially of the guanidinium salt, the buffer and the detergent. 
     
     
         188 . A process for collecting and stabilizing a blood sample for quantitative nucleic acid analysis, the process comprising:
 providing a vessel comprising an interior, the interior comprising an aqueous solution that comprises a guanidinium salt, a buffer and a detergent; and   drawing the blood sample from a blood vessel directly into the interior, wherein the aqueous solution causes lysis of cells in the blood and stabilizes nucleic acids in the sample.   
     
     
         189 . The process of  claim 188 , wherein the interior is evacuated and wherein the evacuation is used to draw the blood sample. 
     
     
         190 . The process of  claim 188 , wherein the vessel comprises a tube having an open end sealed by a septum. 
     
     
         191 . The process of  claim 188 , wherein the evacuation is effective for drawing a specific volume of the blood into the interior. 
     
     
         192 . The process of  claim 188 , wherein the guanidinium salt, the buffer and the detergent are present in amounts effective to provide cell lysis and stabilization of nucleic acids in the sample by inactivation of nucleases. 
     
     
         193 . The process of  claim 188 , wherein the buffer is effective, upon drawing the blood into the interior, to provide the resultant mixture with a pH of 5.0 to 7.6. 
     
     
         194 . The process of  claim 193 , wherein the buffer is effective, upon drawing the blood into the interior, to provide the resultant mixture with a pH of 6.3 to 6.9. 
     
     
         195 . The process of  claim 191 , wherein the specific volume is about 0.1 to about 4 times the volume of the solution. 
     
     
         196 . The process of  claim 188 , wherein the guanidinium salt is one or more salts selected from the group consisting of guanidinium thiocyanate and guanidinium chloride. 
     
     
         197 . The process of  claim 188 , wherein the buffer is selected from the group consisting of: Tris (tris(hydroxymethyl)aminomethane), HEPES (4-(2-hydroxyethyl)-1-piperazineethane-sulfonic acid), MOPS (4-morpholinepropanesulfonic acid), MES (4-morpholineethanesufonic acid), citrate and phosphate buffer. 
     
     
         198 . The process of  claim 188 , wherein the detergent is selected from the group consisting of: (polyethylene glycol tert-octylphenyl ether), (polyethylene glycol 4-nonylphenyl ether), Polydocanol (dodecyl polyethylene glycol ether) and (polyethylene glycol sorbitan monolaurate). 
     
     
         199 . The process of  claim 188 , wherein the pH of the aqueous solution in the vessel prior to the drawing step is 4.0 to 7.5. 
     
     
         200 . The process of  claim 199 , wherein the pH of the aqueous solution in the vessel prior to the drawing step is 5 to 6. 
     
     
         201 . The process of  claim 188 , further comprising the step of engaging the vessel with a blood sampling accessory. 
     
     
         202 . The process of  claim 188 , wherein the solution comprises a single buffer and a single detergent. 
     
     
         203 . The process of  claim 188 , wherein the aqueous solution consists essentially of the guanidinium salt, the buffer and the detergent. 
     
     
         204 . The process of  claim 188 , further comprising the step of isolating the nucleic acids after the drawing step. 
     
     
         205 . The process of  claim 204 , wherein the isolating step is performed at least 3 days after the drawing step. 
     
     
         206 . The process of  claim 205 , wherein the isolating step is performed at least 6 days after the drawing step. 
     
     
         207 . The process of  claim 206 , wherein the isolating step is performed at least 8 days after the drawing step.