Diagnosis of tuberculosis and other diseases using exhaled breath

Abstract

Disclosed are methods and devices for analyzing exhaled breath aerosols and exhaled breath condensates using various diagnostic tools that enable rapid, low cost and autonomous point of care assays for several diseases including respiratory tract diseases. Disclosed are methods and devices for analyzing exhaled breath aerosols and exhaled breath condensates for tuberculosis diagnosis using mass spectrometry, including MALDI-MS. 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 . (canceled) 
     
     
         2 . The system of  claim 51  wherein the EBA particles comprises at least one of microbes, virus, metabolite biomarkers, lipid biomarkers, and proteomic biomarkers characteristic of the respiratory disease. 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . The system of  claim 51  wherein the volume of the collected sample is between about 100 microliter and about 1 ml. 
     
     
         6 . The system of  claim 51  wherein the sample capture component further comprises an air pump, and an impactor wherein the air pump provides the flow of air to carry the exhaled breath from the extraction component into the impactor and wherein the impactor separates the EBA particles from exhaled breath to produce the collected sample. 
     
     
         7 . The system of  claim 6  wherein the impactor comprises at least one of a cyclone, a wetted wall cyclone, one or more wetted film impactors, and an impinger. 
     
     
         8 . The system of  claim 6  further comprising at least one virtual impaction stage disposed upstream of the impactor. 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . The system of  claim 51  wherein the diagnostic device comprises at least one of PCR, rt-PCR, immuno-based assay, mass spectrometer (MS), MALDI-MS, ESI-MS, GC-MS, GC-IMS and MALDI-TOFMS. 
     
     
         12 . (canceled) 
     
     
         13 . The system of  claim 51  wherein the sample capture component is chilled to a temperature greater than about 0° C. and less than about 10° C. using the one or more chilling devices. 
     
     
         14 - 18 . (canceled) 
     
     
         19 . The system of  claim 51  further comprising a sterilization component to disinfect the sample collection subsystem. 
     
     
         20 . The system of  claim 19  wherein the sterilization component comprises at least one of a nebulizer for spraying a disinfectant, one or more UV lights to produce UV radiation, a steam generator, an ozone generator, a peroxide vapor generator, and a combination thereof 
     
     
         21 . The system of  claim 20  wherein the disinfectant comprises at least one of 60% ethanol in water, at least 60% isopropanol in water, and a peroxide solution. 
     
     
         22 . The system of  claim 51  wherein the collected sample is transferred to the sample processing component using at least one of a dispensing pump, gravity-induced flow, and a robotic sample transfer system. 
     
     
         23 - 26 . (canceled) 
     
     
         27 . The system of  claim 51  wherein the sample processing component further comprises at least one of a fluid reservoir, and a fluid dispensing pump to dispense about 1 microliter of fluid on the collected sample disposed on the sample substrate. 
     
     
         28 . The system of  claim 27  wherein the liquid comprises at least one of a solvent, a MALDI matrix chemical, water, and an acid. 
     
     
         29 - 33 . (canceled) 
     
     
         34 . An autonomous method for diagnosing of respiratory diseases in an individual using exhaled breath, the method comprising:
 extracting EBA particles expelled from the individual during a predetermined number of breath maneuvers into a flow of air fed into a sample extraction component configured to receive an individual's face;   collecting the EBA particles from exhaled breath and air as a collected sample using a sample capture component fluidly connected to the sample extraction component by an interface tubing;   spotting a small amount of the collected sample on a sample plate;   processing the sample by treating the sample using at least one of a solvent, a MALDI matrix chemical, water, and an acid, and mixtures thereof; and,   analyzing the sample using a diagnostic device.   
     
     
         35 . The method of  claim 34  wherein the EBA particles comprises at least one of microbes, virus, metabolite biomarkers, lipid biomarkers, and proteomic biomarkers characteristic of the respiratory disease. 
     
     
         36 . The method of  claim 34  wherein the processing step further comprises concentrating the sample by drying the sample using suitable drying means. 
     
     
         37 . The method of  claim 34  wherein the diagnostic device comprises MALDI-TOFMS. 
     
     
         38 . (canceled) 
     
     
         39 . The method of  claim 34  further comprising the step of digesting the collected sample to generate a peptide sample characteristic of the EBA particles. 
     
     
         40 - 42 . (canceled) 
     
     
         43 . The method of  claim 34  wherein the flow rate of air entering the sample capture component is between about 100 L/min and about 1000 L/min. 
     
     
         44 - 45 . (canceled) 
     
     
         46 . The method of  claim 34  wherein the predetermined number of breath maneuvers comprises the following steps:
 a. performing a deep exhale to clear the individual's lungs; 
 b. pausing for up to 10 s; 
 c. performing an FVC inhale; 
 d. performing a deep exhale; and, 
 e. repeating the above sequence for up to 10 times. 
 
     
     
         47 . The method of  claim 46  further comprising at least one of the steps of tidal breathing, coughing, normal FVC breaths, speaking, and sneezing. 
     
     
         48 . (canceled) 
     
     
         49 . The method of  claim 34  wherein the number of pre-determined breath maneuvers is determined by one or more sensors that indicate at least one of the volume of particles exhaled and the volume of breath exhaled. 
     
     
         50 . The method of  claim 49  wherein the one or more sensors comprises at least one of a CO 2  sensor, an oxygen sensor, a humidity sensor, an optical particle size counter, an aerodynamic particle sizer, and a nephelometer. 
     
     
         51 . A system for diagnosis of a respiratory disease using exhaled breath, the system comprising:
 a sample capture component configured to collect EBA particles in a predetermined volume of air fed into the sample capture component as a collected sample wherein the air flow is between about 30 L/min and about 1000 L/min; and,   a sample analysis subsystem fluidly connected to the sample capture component, the sample analysis subsystem comprising:
 a sample processing component to spot a small amount of the collected sample on a sample plate and treat the collected sample on the sample plate; and, 
 a diagnostic device for analyzing the sample. 
   
     
     
         52 . The system of  claim 51  wherein the sample processing component comprises fluidic components to treat the sample using at least one of a solvent, a MALDI matrix chemical, water, and an acid, and mixtures thereof. 
     
     
         53 . The system of  claim 51  further comprising one or more sensors configured be in fluid communication with the sample extraction component wherein the output of the one or more sensors is used to calculate the total cumulative volume of exhaled breath aerosol particles entering the sample capture component. 
     
     
         54 . The system of  claim 53  wherein the one or more sensors comprises at least one of a CO 2  sensor, an oxygen sensor, a humidity sensor, an optical particle size counter, an aerodynamic particle sizer, and a nephelometer. 
     
     
         55 . The system of  claim 51  wherein the predetermined volume of air is determined using the output of the one or more sensors.