Magnetic particles with a closed ultrathin silica layer, method for the production thereof and their use
A method for producing silicate-containing magnetic particles having a closed and tight silicate layer and high purity. In addition, the novel method prevents an uncontrolled formation of aggregates and clusters of silicates on the magnetite surface, thereby having a positive influence on the properties and biological applications. The method enables depletion of nanoparticulate solid substance particles on the basis of a fractionated centrifugation. The silicate-coated magnetic particles exhibit optimized magnetization and suspension behavior as well as advantageous run-off behavior from plastic surfaces. These highly pure magnetic particles coated with silicon dioxide are preferably used for isolating nucleic acids from cell and tissue samples, whereby the separating out from a sample matrix ensues by means of magnetic fields. The particles are particularly well suited for the automatic purification of nucleic acids, mostly from biological body samples for the purpose of analyzing them with different amplification methods.
1. A method of purification of nucleic acid from a biological sample, comprising:
dissolving the biological sample containing a nucleic acid of HCV or HIV into a sample solution;
forming silica-coated magnetic particles by depositing initial-silicate layers on magnetic particles and continuously diluting using cross-flow microfiltration to reduce a pH value to a neutral pH value;
adding the silica-coated magnetic particles to the sample solution;
incubating the sample solution at a temperature at which the nucleic acid bonds to the silica-coated magnetic particles;
applying, at a first time, a first magnetic field to the sample solution;
removing constituents not bonded to the silica-coated magnetic particles;
removing unspecifically bonded molecules from the nucleic acid by applying and removing a washing buffer;
separating nucleic acid from the silica-coated magnetic particles by adding an elution buffer; and
applying, at a second time, a second magnetic field and separating eluate with the nucleic acid of HCV or HIV.
2. The method of claim 1 , wherein the biological sample is infected with HCV.
3. The method of claim 1 , wherein the biological sample is infected with HIV.
4. The method of claim 1 , wherein the nucleic acid is RNA of HCV or HIV.
5. The method of claim 1 , wherein prior to continuously diluting, carrying out a fractionated centrifugation.
6. A method for purification of nucleic acids from a biological sample in viral diagnostics, comprising:
dissolving the biological sample containing nucleic acid of HCV or HIV into a sample solution;
forming silica-coated magnetic particles by depositing initial-silicate layers on magnetic particles and continuously diluting using cross-flow microfiltration to reduce a pH value to a neutral pH value;
adding the silica-coated magnetic particles to the sample solution;
incubating the sample solution at a temperature at which the nucleic acid bonds to the silica-coated magnetic particles;
applying a magnetic field to the sample solution;
removing constituents not bonded to the silica-coated magnetic particles;
removing unspecifically bonded molecules from the nucleic acid by applying and removing a washing buffer a plurality of times;
separating the nucleic acid from the silica-coated magnetic particles by adding an elution buffer;
applying another magnetic field and separating eluate with the nucleic acid of HCV and HIV; and
analyzing nucleic acid of HCV or HIV obtained from eluate.
7. The method of claim 6 , further comprising:
prior to adding the silica-coated magnetic particles to the sample solution, carrying out a fractionated centrifugation on the silica-coated magnetic particles; and
subjecting the nucleic acid obtained from the eluate to an amplification reaction prior to analyzing.
8. The method of claim 7 , wherein the amplification reaction is selected from a group consisting of a polymerase chain reaction, a transcription mediated amplification, a ligase chain reaction, and a nucleic acid sequence based amplification.
9. The method of claim 7 , wherein the nucleic acid obtained from the amplification reaction includes nucleic acid of HCV.
10. The method of claim 7 , wherein the nucleic acid obtained from the amplification reaction includes nucleic acid of HIV.