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JP-7855248-B2 - Diagnosis of respiratory diseases using exhaled breath and aerosol analysis

JP7855248B2JP 7855248 B2JP7855248 B2JP 7855248B2JP-7855248-B2

Inventors

  • チェン、ダペン
  • ブライデン、ウェイン、エイ．
  • マクローリン、マイケル

Assignees

  • ゼテオ テック、 インク．

Dates

Publication Date
20260508
Application Date
20240605
Priority Date
20200403

Claims (14)

  1. In a system for detecting protein-containing aerosol particles in exhaled breath, An exhaled breath collection element configured to accept an individual's face in order to collect aerosol particles containing proteins in exhaled breath, wherein the exhaled breath collection element fits snugly to the individual's face, When the above exhaled breath collection element is placed on the face of the individual, the exhaled breath collection element includes a port located near the individual's jaw, A sample capture element comprising a packed bed column for selectively capturing the above-mentioned aerosol particles containing protein, wherein the sample capture element is detachably connected to the port without interconnecting tubes, A pump configured to communicate with the above-mentioned sample capture element and draw exhaled air into the above-mentioned sample capture element, A sample extraction system configured to extract protein particles from the packed bed column described above, A sample processing system comprising means for high-temperature acid digestion of the protein particles extracted from the sample extraction system in order to generate a peptide sample characteristic of the captured protein, A system characterized by having a diagnostic device for analyzing the above-mentioned peptide sample.
  2. The system according to claim 1, wherein the protein particles include proteins related to a virus, comprising one or more of SARS-CoV, MERS-CoV, or SARS-CoV-2.
  3. The system according to claim 1, wherein the above-mentioned protein comprises a lower respiratory tract-derived protein containing one or more of the following: serum albumin, keratin, glycoprotein, cystatin, dermacidine, or S100 protein.
  4. The system according to claim 1, wherein the particle capture efficiency of the sample capture element is greater than 99%.
  5. The system according to claim 1, wherein the exhaled breath collection element includes one or more of the following: a CPR rescue mask, a CPAP mask, or a ventilator mask.
  6. The system according to claim 1, wherein the packed bed column comprises resin beads containing C18 functional groups on its surface.
  7. The system according to claim 1, wherein the packed bed column comprises heparin covalently bonded to Sepharose beads.
  8. The packed bed column comprises solid particles on which a functional group is immobilized on the surface of the particles, wherein the functional group comprises one or more carbohydrates including glycan, heparin, heparan sulfate, or dextran, according to claim 1.
  9. The system according to claim 8, wherein the above-mentioned solid particles are packed between two porous polymer frit disks.
  10. The extraction system according to claim 1, comprising means of flushing the packed bed column with one or more of the following: about 12.5% acetic acid, about 5% TFA, about 5% formic acid, about 70% isopropanol, or about 10% HCl.
  11. The system according to claim 1, wherein the pump includes a diaphragm pump.
  12. The diagnostic device described above is the system according to claim 1, comprising one or more of the following: PCR, ELISA, rt-PCR, mass spectrometer (MS), LC-MS, MALDI-MS, ESI-MS, or MALDI-TOFMS.
  13. The system according to claim 1, further comprising a trap positioned between the sample capture element and the pump, configured to capture exhaled condensate (EBC) containing one or more of water vapor, volatile organic components, or non-volatile organic components passing through the packed bed.
  14. The system according to claim 13, wherein the trap is cooled to a temperature lower than the ambient temperature.

Description

This disclosure relates to methods and devices for analyzing non-volatile organic compounds in exhaled breath and other aerosols using a variety of diagnostic tools that enable rapid, low-cost, and autonomous point-of-care assays for respiratory diseases. More specifically, but not limited to, this disclosure relates to methods and devices for analyzing non-volatile organic compounds in exhaled breath using mass spectrometry methods, including MALDI-TOFMS, for the detection of respiratory diseases such as COVID-19 and for the diagnosis of tuberculosis. Coronavirus disease (COVID-19) is a disease caused by the newly emerged coronavirus SARS-CoV-2. This new coronavirus is a respiratory virus that spreads primarily through droplets produced when an infected person coughs or sneezes, or through saliva droplets or nasal secretions. The new coronavirus is highly contagious and has caused the ongoing COVID-19 pandemic. This suggests that this virus is spreading more rapidly than influenza. Rapid detection tools are needed to help mitigate the spread. Furthermore, tuberculosis (TB) kills more than 4,000 people daily, surpassing HIV/AIDS as the leading cause of death (killer) globally (Patterson, B et al., 2018). The reported rate of incidence reduction remains insufficient at 1.5% per year, making it unlikely that treatment alone will significantly reduce the disease burden. In HIV-prevalent communities, genotyping studies of Mycobacterium tuberculosis (Mtb) have shown that recent infections, rather than relapses, account for the majority (54%) of tuberculosis cases. The physical processes of tuberculosis transmission remain poorly understood, and the application of new technologies to elucidate key events in the generation, release, and inhalation of infectious aerosols is slow. There are few empirical studies characterizing airborne infectious particles. Two major problems hindering investigations are the low concentrations of naturally generated Mtb particles and the complications of environmental and patient-derived bacterial and fungal contamination of airborne samples. Nevertheless, many attempts at airborne detection have been made. A 2004 proof-of-concept study in Uganda and subsequent feasibility studies sampled aerosols generated by coughing from pulmonary tuberculosis patients. Direct coughing into a sampling chamber equipped with two viable cascade impactors yielded positive cultures in more than a quarter of participants, despite 1–6 days of chemotherapy. Follow-up studies using the same apparatus found that participants with high aerosol bacterial loads had higher rates of household infections, potentially leading to disease detection that quantitative airborne sampling could serve as a useful measure of clinically relevant infectivity. Therefore, interruptions in transmission may have a rapid and measurable impact on tuberculosis incidence. The best way to control the transmission of tuberculosis is to rapidly identify and treat active TB cases (Wood, R.C. et al., 2015). Diagnosis of pulmonary tuberculosis is usually made by microbiological, microscopic, or molecular analysis of the patient's sputum. The “gold standard” test for tuberculosis infection in most developing countries is smear culture based on sputum samples. The sample is smeared onto a culture plate, a stain specific to Mtb is added, and the stained cells are counted using a microscope. If the cell concentration in the smear is higher than a set threshold, the sample is classified as positive. If the TB count is below this threshold, it is classified as negative. Diagnosis can take several hours. The need for sputum as a diagnostic sample is limited by the challenge of collecting sputum from patients and its complex composition. The viscosity of the material limits the sensitivity of the test, increases heterogeneity between samples, and increases the cost and effort associated with the test. Furthermore, sputum (which requires coughing) poses an occupational risk to healthcare workers. Sputum has several drawbacks as a sample medium. First, only about 50% of patients can provide a good sputum sample. For example, children around the age of eight often cannot provide a sample when requested because they have not yet developed the ability to "cough" sputum from the back of their throat. The elderly and sick may not have the strength to cough up phlegm. Others may simply not have phlegm in their throat. Therefore, diagnostic methods based on sputum analysis may not be able to provide a diagnosis for as many as 50% of patients who need one. Sputum is not useful as a diagnostic sample if it is collected one to two days after the patient has been treated with antibiotics. This is because the specimen no longer represents the disease deep in the lungs, and the number of living Mtb in the sputum decreases significantly within a few days of starting treatment. Urine and blood have been proposed as sample mediums for the diagnosis of tuberculosi