| 摘要 |
<p>#CMT# #/CMT# Method for decontamination of solution containing nucleic acid and its contaminant, comprises: subjecting the solution to action of a molecule having property to degrade nucleic acids by fragmentation and the molecule for fragmentation, until the total degradation of the nucleic acids and its contaminants while retaining the desired nucleic acids; and arresting the activity of the molecule for fragmentation. #CMT#USE : #/CMT# The method is useful in diagnostic field. #CMT#ADVANTAGE : #/CMT# The method is easy to carry out and avoids the contamination of new samples. #CMT#BIOLOGY : #/CMT# Preferred Components: The solution contains all constituents necessary for amplification reaction except nucleic acids, either targets or amplification primers and probe for detection. The treated nucleic acids are RNA or DNA in the form of single or double strand and RNA/DNA heteroduplex. The duration of the treatment by treated nucleic acids is 5-60 minutes, preferably 10-30 minutes for the concentrations of ClipPhen/metal (1-100 mu M). #CMT#ORGANIC CHEMISTRY : #/CMT# Preferred Method: The method comprises an additional step comprising an amplification reaction, after arresting step. The method comprises subjecting the solution to the action of ClipPhen/metal molecule, until total degradation of the nucleic acids and its contaminants, while preserving the desired nucleic acids. The solution is mixed with a biological sample containing target nucleic acids for amplifying, but also primers for amplification and probes for specific detection of target nucleic acids in the presence of a second organic reducer, for creating an excess of organic reducer, which arrests the action of molecules for fragmentation. Preferred Components: The molecule for fragmentation is: used for randomly hydrolyzing the diester phosphate bonds of nucleic acid contaminants; a complex constituted of two molecules of 1,10-phenanthroline associated with a metal atom forming the complex bis (1,10-phenanthroline)/metal, preferably transition metal comprising copper, ruthenium, nickel, iron, zinc, rhodium, cobalt and manganese. The two phenanthroline core of the molecule are linked to one another by a bonding arm to form a ClipPhen molecule. The arm of bonding between two phenanthroline cores is constituted by a chain of three successive carbon atoms, where the carbon atom is substituted in central position and each carbon atom of carbon terminal is linked to the phenanthroline core by an oxygen atom. The carbon atom is substituted by amine or acetamide, and that each atom of carbon terminal is linked to the phenanthroline core by oxygen atom in position 2 or 3 of the core. The ClipPhen molecule is 3-ClipPhen (1,3-bis (1,10-phenanthroline-3-yloxy)-propan-2-amino). The molecule comprises: a type I copper complex associated with hydrogen peroxide and optionally with a reducer, preferably organic; and a type II copper complex preferably associated with an organic reducer. The reducer is an organic reducer comprising thiol comprising: dithiothreitol, thio-acid and mercapto-propionic acid; or carboxylic acid; or carboxylic acid derivative comprising ascorbate; or phosphine as tri carboxy ethyl phosphine. The molecule is immobilized on a solid support. The reducer is inactive on a solid support. The solid support is particle, membrane, bandage or filter. The ratio between the molecule and organic reducer constituted by carboxylic acid or derivative is 1-1 and 1-100. The fragmentation activity of the molecule is arrested by an addition of an excess organic reducer or a complexing agent (EDTA). The ratio between the molecule and thiol is greater than 1-100, preferably greater than 1-1000. The fragmentation activity of the molecule is arrested when the solid support is removed from the solution. The second organic reducer is identical to the first organic reducer or different from first organic reducer. #CMT#EXAMPLE : #/CMT# No suitable example given.</p> |
