Method of amplifying nucleic acid from a cell using a nonplanar solid substrate
Provided is a method of amplifying nucleic acid from a comprising: contacting a cell-containing sample with a nonplanar solid substrate in a liquid medium having a pH range of 3.0-6.0 to attach the cell to the solid substrate; washing the nonplanar solid substrate to remove materials that are not attached thereto; and performing PCR using the nucleic acid from the cell attached to the nonplanar solid substrate as a template sample to amplify nucleic acid from the cell, wherein the contacting, washing and PCR processes are performed in a single vessel.
1. A method of amplifying nucleic acid from a cell comprising:
contacting a sample comprising a cell and polyanethole sulfonate with a nonplanar solid substrate in a liquid medium having a pH in a range of 3.0-6.0 to attach the cell to the nonplanar solid substrate,
wherein the nonplanar solid substrate is hydrophobic and has a water contact angle of 70° to 95° or the nonplanar solid substrate has at least one amine-based functional group at its surface prepared by coating the nonplanar solid substrate with polyethyleneiminetrimethoxysilane (PEIM);
washing the nonplanar solid substrate to remove materials that are not attached thereto; and
performing a polymerase chain reaction (PCR) using the cell attached to the nonplanar solid substrate as a template sample to amplify nucleic acid from the cell,
wherein the contacting, washing and performing the PCR are performed in a single vessel and the cell is a microorganism cell.
2. The method of claim 1 , wherein the sample is blood, blood culture, urine or saliva.
3. The method of claim 1 , wherein the cell is a bacterial cell, a fungus, or a virus.
4. The method of claim 1 , wherein the sample is diluted in the liquid medium, which is a phosphate buffer or an acetate buffer.
5. The method of claim 4 , wherein the sample is diluted in a ratio of 1:1 to 1:10.
6. The method of claim 5 , wherein the liquid medium has a salt concentration of 10 mM to 500 mM.
7. The method of claim 6 , wherein the sample has a salt concentration of 50-300 mM.
8. The method of claim 1 , wherein the nonplanar solid substrate is a solid substrate comprising a surface with a plurality of pillars, a bead-shaped solid substrate, or a sieve-shaped solid substrate having a plurality of pores.
9. The method of claim 8 , wherein the pillars have an aspect ratio of 1:1-20:1, wherein the aspect ratio refers to a ratio of the cross-sectional diameter of a pillar to the height of a pillar.
10. The method of claim 8 , the plurality of pillars comprises a pillar structure with a ratio of a height of the pillars to a distance between adjacent pillars in the range of 1:1 to 25:1.
11. The method of claim 8 , wherein the plurality of pillars comprises a pillar structure and a distance between adjacent pillars is in the range of 5 μm to 100 μm.
12. The method of claim 1 , wherein the hydrophobic nonplanar solid substrate is obtained by coating octadecyldimethyl(3-trimethoxysilyl propyl)ammonium (OTC) or tridecafluorotetrahydrooctyltrimethoxysilane (DFS) on a surface of the nonplanar solid substrate.