Diagnosis of respiratory diseases using analysis of exhaled breath and aerosols
Disclosed are methods and devices for analyzing non-volatile organics in exhaled breath and other aerosols using various diagnostic tools that enable rapid, low cost point of care assays for several diseases including respiratory tract diseases such as COVID-19. The disclosed methods and systems selectively capture non-volatile organics in exhaled breath and other aerosols in a packed bed column. The non-volatile organics are eluted and samples are analysis using diagnostic devices including MALDI-TOFMS. The disclosed systems and methods provide for a diagnostic test result in less than about 20 minutes and provides for autonomous operation with minimal human intervention.
1. A breath sample collection system for diagnosis of at least one respiratory disease using exhaled breath, the system comprising:
a breath collection element configured to receive an individual's face for collecting aerosolized bacteria and virus particles in exhaled breath wherein the breath collection element forms a tight-fit with the individuals face;
a sample capture element comprising a packed bed column to selectively capture the aerosolized bacteria and virus particles wherein the sample capture element is removably connected to a port disposed in the breath collection element proximate to the individual's chin when the breath collection element is positioned on the individual's face without any interconnecting tubing; and,
a pump in fluid communication with the sample capture element and configured to draw exhaled breath into the sample capture element wherein the particle capture efficiency of the breath sample collection system is greater than 99%.
2. The system of claim 1 wherein the packed bed column comprises solid particles comprising at least one of resins, cellulose, silica, agarose, and hydrated Fe 3 O 4 nanoparticles.
3. The system of claim 2 wherein the solid particles comprise functional groups immobilized on the surface of the particles wherein the functional groups comprise at least one of C18 (octadecyl), octyl, ethyl, cyclohexyl, phenyl, cyanopropyl, aminopropyl, 2,3-dihydroxypropoxypropyl, trimethyl-aminopropyl, carboxypropyl, benzenesulfonic acid, propylsulfonic acid, an ion exchange phase, a polymer phase, antibodies, glycans, lipids, DNA and RNA.
4. The system of claim 1 wherein the packed bed column comprises resin beads having C18 functional groups on the surface.
5. The system of claim 4 wherein the resin beads have a nominal diameter of between about 12 μm and about 20 μm.
6. The system of claim 1 wherein the nominal flow rate pulled through the packed bed column by the pump is between about 200 ml/min and about 600 ml/min.
7. The system of claim 1 wherein the breath collection element comprises at least one of a CPR rescue mask, a CPAP mask, and a ventilator mask.
8. The system of claim 1 further comprising a trap disposed between the sample capture element and the pump and configured to trap exhaled breath condensate (EBC) comprising at least one of water vapor, volatile organic components and non-volatile organic components that pass through the packed bed wherein the trap is cooled below ambient temperature.
9. The system of claim 1 wherein the capture element is cooled to a temperature at or below ambient temperature.
10. A system for diagnosis of a respiratory disease using exhaled breath, the system comprising:
the breath sample collection system of claim 1 ;
a sample extraction system to extract captured virus and bacteria particles from the packed bed column; and,
a sample analysis system comprising:
a sample processing system for treating and concentrating the collected sample on a sample plate; and,
a diagnostic device for analyzing the sample.
11. The system of claim 10 wherein the diagnostic device comprises at least one of PCR, ELISA, rt-PCR, mass spectrometer (MS), MALDI-MS, ESI-MS, and MALDI-TOFMS.
12. The system of claim 10 wherein the extraction system comprises means to flush the pack bed column with a solvent and remove the solvent comprising aerosolized bacteria and virus particles from the packed bed.
13. The system of claim 12 wherein the solvent comprises at least one of acetonitrile, methanol, acid, isopropanol, the remaining being water.
14. The system of claim 10 wherein the sample processing system comprises:
means for hot acid digestion of the bacteria and virus particles extracted from the sample extraction system to generate a peptide sample characteristic of the particles;
mixing the peptide sample with a MALDI matrix; and,
applying the mixed sample and MALDI matrix to a sample plate.
15. A sample capture element for diagnosis of a respiratory disease using exhaled breath comprising:
a packed bed column to selectively capture aerosolized bacteria and virus particles in exhaled breath drawn through the packed bed column using a pump wherein the packed bed column comprises:
solid particles having a nominal diameter of between about 12 μm and about 20 μm comprising at least one of resins, cellulose, silica, agarose, and hydrated Fe 3 O 4 nanoparticles; and,
functional groups immobilized on the surface of the particles wherein the functional groups comprise at least one of C18 (octadecyl), octyl, ethyl, cyclohexyl, phenyl, cyanopropyl, aminopropyl, 2,3-dihydroxypropoxypropyl, trimethyl-aminopropyl, carboxypropyl, benzenesulfonic acid, propylsulfonic acid, ion exchange phases, polymer phases, antibodies, glycans, lipids, DNA and RNA,
wherein the particle capture efficiency of the sample capture element is greater than 99% as the exhaled breath is drawn through the packed bed column at a flow rate of between about 200 ml/min and about 600 ml/min.
16. The element of claim 15 wherein the solid particles are packed between two porous polymeric frit discs.
17. A breath sample collection system for diagnosis of at least one respiratory disease using exhaled breath, the system comprising:
a mask configured to receive an individual's face for collecting aerosolized bacteria and virus particles in exhaled breath wherein the mask forms a tight-fit with the individuals face and wherein the mask comprises a stem and port disposed below the stem proximate to the individual's chin when the mask is positioned on the individual's face;
a HEPA filter removably and fluidly connected to the stem of the mask;
a sample capture element comprising a packed bed column to selectively capture the aerosolized bacteria and virus particles wherein the sample capture element is removably connected to the port; and,
a pump in fluid communication with the sample capture element and configured to draw exhaled breath into the sample capture element wherein the particle capture efficiency of the sample capture element is greater than 99% as the exhaled breath is pulled through the packed bed column by the pump at a flow rate of between about 200 ml/min and about 600 ml/min.
18. The system of claim 17 wherein the packed bed column comprises solid particles comprising at least one of resins, cellulose, silica, agarose, and hydrated Fe 3 O 4 nanoparticles.
19. The system of claim 17 wherein the packed bed column comprises resin beads having C18 functional groups on the surface.
20. The system of claim 19 wherein the resin beads have a nominal diameter of between about 12 μm and about 20 μm.