Rapid nucleic acids separation and sample preparation via hollow-centered silica microsphere

A method for amplifying a nucleic acid separated from a sample, the method comprising:

• separating the nucleic acid from the sample;
• wherein separating the nucleic acid from the sample comprises:
• (a) contacting a sample lysate with a plurality of nucleic-acid-capture microspheres within a microsphere-separating device comprising a microsphere retaining mesh to form a lysate dispersion;
• wherein the lysate dispersion comprises a lysate continuous phase and a particulate phase and wherein the particulate phase comprises the plurality of nucleic-acid-capture microspheres and an adsorbed nucleic acid obtained from the sample;
• (b) mechanically separating the lysate continuous phase from the particulate phase comprising the plurality of nucleic-acid-capture microspheres and the adsorbed nucleic acid obtained from the sample;
• and
• (c) contacting the particulate phase with an eluent to form an eluate comprising the nucleic acid obtained from the sample;
• contacting the nucleic acid separated from the sample with an amplification medium;
• and
• amplifying the nucleic acid separated from the sample,
• wherein the plurality of nucleic-acid-capture microspheres comprises soda-lime-borosilicate microspheres.

A method of detecting a nucleic acid in a sample indicative of a pathogen or a cell, the method comprising:

• separating the nucleic acid from the sample, wherein separating the nucleic acid from the sample comprises:
• (a) contacting a sample lysate with a plurality of nucleic-acid-capture microspheres within a microsphere-separating device comprising a microsphere retaining mesh to form a lysate dispersion;
• wherein the lysate dispersion comprises a lysate continuous phase and a particulate phase and wherein the particulate phase comprises the plurality of nucleic-acid-capture microspheres and an adsorbed nucleic acid obtained from the sample;
• (b) mechanically separating the lysate continuous phase from the particulate phase comprising the plurality of nucleic-acid-capture microspheres and the adsorbed nucleic acid obtained from the sample;
• and
• (c) contacting the particulate phase with an eluent to form an eluate comprising the nucleic acid obtained from the sample;
• contacting the nucleic acid separated from the sample with an amplification medium, wherein the amplification medium comprises a primer complementary to a target sequence indicative of the pathogen or the cell;
• amplifying the nucleic acid separated from the sample to form an amplified sample comprising a plurality of amplicons of the target sequence;
• and
• detecting the target sequence indicative of the pathogen or the cell,
• wherein the plurality of nucleic-acid-capture microspheres comprises soda-lime-borosilicate microspheres.