Processing blood samples to detect target nucleic acids

1. A method of fabricating a porous polymer monolith, comprising:

providing a plurality of monomers comprising:

at least one monomer; and

at least one radically polymerizable trimethylolpropane (TMP)-based monomer having the formula:

wherein R′, R 2 , and R 3 are each independently selected from —(CH 2 CH 2 O) n ,

—(CH 2 CH 2 O) n C(O)CH 2 CH 2 SH, —(CH 2 CH 2 O) n C(O)CH═CH 2 , and

—(CH 2 CH 2 O) n C(O)C(CH 3 )═CH 2 ; and each n is independently an integer from 0 to 12:

and wherein the at least one TMP-based monomer comprises trimethylolpropane tris(3-mercaptopropionate) (TMPMP) in an amount ranging from 0.1% to 7% (v:v) of the total volume of the plurality of monomers

obtaining a polymerizable composition by combining the plurality of monomers in a porogenic solvent; and

polymerizing the polymerizable composition to form the porous polymer monolith.

2. The method of claim 1 , wherein n is 0.

3. The method of claim 1 , wherein the at least one monomer comprises trimethylolpropane ethoxy triacrylate (TMP(EO)TA) in an amount ranging from 0.1% to 44% (v:v) of the total volume of the plurality of monomers.

4. The method of claim 1 , wherein the at least one monomer is selected from ethylene glycol dimethacrylate (EGDMA);

2-hydroxyethyl methacrylate (HEMA);

tetra(ethylene glycol) diacrylate (TEGDA); and

tetra(ethylene glycol) dimethacrylate (TEGDMA); or a combination thereof.

5. The method of claim 1 , wherein the at least one monomer is selected from

ethylene glycol dimethacrylate (EGDMA) in an amount ranging from 34% to 75% (v:v) of the total volume of the plurality of monomers;

2-hydroxyethyl methacrylate (HEMA) in an amount ranging from 10% to 35% (v:v) of the total volume of the plurality of monomers;

tetra(ethylene glycol) diacrylate (TEGDA) in an amount ranging from 0% to 15% (v:v) of the total volume of the plurality of monomers; and

tetra(ethylene glycol) dimethacrylate (TEGDMA) in an amount ranging from 0% to 20% (v:v) of the total volume of the plurality of monomers.

6. The method of claim 1 , wherein the porogenic solvent comprises at least one of the following

alcohols;

a mixture of alcohols and water;

a first alcohol of the formula: [C X H (2X+2) O], wherein X is an integer from 1 to 10; and

a second alcohol of the formula: [C Y H (2Y+2) O 2 ], wherein Y is an integer from 2 to 10.

7. The method of claim 1 , wherein the porogenic solvent further comprises at least one of the following:

a surfactant;

sodium dodecyl sulfate (SDS),

polyethylene-polypropylene glycol, and

polyethylene glycol tert-octylphenyl ether.

8. The method of claim 1 , wherein the polymerizable composition has a monomer to solvent ratio of 1:1 to 1:5 (v:v).

9. The method of claim 1 , wherein the monolith is a self-wicking monolith having a two minute water wick rate between 1.5 and 5.0 centimeters.

10. The method of claim 1 , wherein the monolith has a measured red blood cell retention factor (Rf) value in the range of 0.01 to 0.8.

11. The method of claim 1 , wherein the monolith has a pore size within a range of 2-7 microns and a porosity of 50 to 85 percent.

12. The method of claim 1 , wherein the monolith is a lateral flow monolith.

13. The method of claim 1 , wherein the monolith is a lateral flow porous monolith for the fractionation of a blood sample into blood cells and a blood-cell-free fraction.

14. The method of claim 1 , wherein the monolith has a minimum tensile strength corresponding to a supported weight of at least 10 grams.

15. A monolith fabricated by the method of claim 1 .