Periodic Reporting for period 1 - B-Path (Breath Pathogen Detection (B-Path): Establishing Exhaled Breath Aerosol (XBA) sampling for diagnosis and screening of respiratory infections)
Période du rapport: 2024-07-01 au 2025-12-31
Respiratory infections resulted in >7 million deaths in 2020 and were responsible for 7 of the last 9 pandemics, causing trillions of €s in economic losses. Despite the importance of early detection for individual health and pandemic control, suboptimal sampling methods for respiratory infections limit the impact of highly-sensitive diagnostics. B-Path will tackle this problem by establishing exhaled breath aerosols (XBA) as an evidence-based, non-invasive sample for simple detection of respiratory infections that can be undertaken at all levels of care. We will achieve this by developing and optimising two innovative, easy-to-use and scalable exhaled breath aerosol sampling devices and generating evidence for their use in diagnosis and screening (for early diagnosis and transmission control).
We will use different model pathogens that have caused epidemics and pandemics globally and generate data on the feasibility of multiplex detection of respiratory pathogens in XBA samples for different use cases, including the ability to detect novel pathogens through metagenomics, and point-of-care testing with lateral-flow assays. We will assess the sampling efficiency of the novel devices against the Respiratory Aerosol Sampling Chamber as a benchmark, and evaluate performance, feasibility and acceptability, in comparison to the current standard of sampling in multi-country clinical studies. Impact and economic modelling will inform the implementation potential of breath sampling devices for the different use cases and help assess trade-offs. We envision that a single breath sample enables accessible and accurate detection of highly transmissible respiratory infections, thereby improving both individual and population health.
We will use different model pathogens that have caused epidemics and pandemics globally and generate data on the feasibility of multiplex detection of respiratory pathogens in XBA samples for different use cases, including the ability to detect novel pathogens through metagenomics, and point-of-care testing with lateral-flow assays. We will assess the sampling efficiency of the novel devices against the Respiratory Aerosol Sampling Chamber as a benchmark, and evaluate performance, feasibility and acceptability, in comparison to the current standard of sampling in multi-country clinical studies. Impact and economic modelling will inform the implementation potential of breath sampling devices for the different use cases and help assess trade-offs. We envision that a single breath sample enables accessible and accurate detection of highly transmissible respiratory infections, thereby improving both individual and population health.
Objective 1: Develop and optimise two breath-based aerosol sampling methods to detect respiratory pathogens with pandemic potential
During this reporting period, consortium members Avelo and Goethe University Frankfurt (GUF) successfully achieved Objective 1 by completing Work Package (WP) 2. This entailed developing, validating, and operationalising two breath-based aerosol sampling technologies (AveloCollect and AveloMask) together with molecular assays for detection. All activities defined under WP2 were achieved, with the only operational adaptation being the shift from single-batch manufacturing of the AveloMask to staged production and shipment aligned with clinical recruitment timelines. The work under Objective 1 has delivered validated breath aerosol sampling technologies, validated extraction and multiplex virus testing workflows, regulatory-ready manufacturing, and decisive performance data guiding downstream implementation. The consortium is now positioned to advance AveloMask, the most effective XBA sampling solution, into the next clinical phases in full alignment with the project’s scientific, clinical and public-health objectives.
Objective 2: Development of RASC as a benchmark for aerosol detection of respiratory pathogens and comparison of 3 different sampling methods
Ethical approval for the B-Path study was granted by the South African Human Research Ethics Council (HREC) following submission of clarifications regarding pre-specified safety inclusion and exclusion criteria, including thresholds for oxygen saturation, pulse, blood pressure, respiratory rate, and dyspnoea. Approval from the City of Cape Town was subsequently received in October 2025. A total of 80 AveloCollect devices and AveloMasks were received for use in the study.
For downstream analysis, the team agreed to proceed with nucleic acid extraction using the QIAsymphony platform, and finalisation of the analytical workflow is happening based on the data generated from WP2. Recruitment for the clinical study is due to commence in March 2025 to co-incide with the South African respiratory viral season.
Objective 3: Evaluate the diagnostic accuracy of XBA sampling in symptomatic adults, and feasibility of XBA sampling for detecting asymptomatic infection
The project’s clinical studies will be conducted under Objective 3, whereby Project B-Path aims to evaluate the diagnostic accuracy of an exhaled breath aerosol sample in symptomatic adults, and feasibility of sampling for detecting asymptomatic infection. The multisite coordinated study protocol has been developed to be implemented in Bucharest (Romania), Milan (Italy) and Heidelberg (Germany). Ethical approvals have been received, the trial registered in the German Clinical Trial Registry and the studies were registered as the different teams prepared each site for recruitment commencement in March 2026.
Objective 4: Evaluate acceptability, usability, population impact, and cost-effectiveness of XBA sampling for diagnosis and screening of respiratory infections of pandemic potential
Initial work on human factors engineering has been completed with some valuable insights gathered around the mask design, packaging, hygiene, acceptability and consideration for adaptation. A mixed-methods study of 22 participants with both AveloCollect and AveloMask lay the groundwork for evaluating the devices during the remainder of the project. Plans to further assess acceptability, appropriateness and usability are nested within the study protocols under Objective 3 and will take place in Germany, Romania and Italy in adults being diagnosed and screened for respiratory infections.
Population impact and cost-effectiveness of XBA will be analysed in the second half of the project.
During this reporting period, consortium members Avelo and Goethe University Frankfurt (GUF) successfully achieved Objective 1 by completing Work Package (WP) 2. This entailed developing, validating, and operationalising two breath-based aerosol sampling technologies (AveloCollect and AveloMask) together with molecular assays for detection. All activities defined under WP2 were achieved, with the only operational adaptation being the shift from single-batch manufacturing of the AveloMask to staged production and shipment aligned with clinical recruitment timelines. The work under Objective 1 has delivered validated breath aerosol sampling technologies, validated extraction and multiplex virus testing workflows, regulatory-ready manufacturing, and decisive performance data guiding downstream implementation. The consortium is now positioned to advance AveloMask, the most effective XBA sampling solution, into the next clinical phases in full alignment with the project’s scientific, clinical and public-health objectives.
Objective 2: Development of RASC as a benchmark for aerosol detection of respiratory pathogens and comparison of 3 different sampling methods
Ethical approval for the B-Path study was granted by the South African Human Research Ethics Council (HREC) following submission of clarifications regarding pre-specified safety inclusion and exclusion criteria, including thresholds for oxygen saturation, pulse, blood pressure, respiratory rate, and dyspnoea. Approval from the City of Cape Town was subsequently received in October 2025. A total of 80 AveloCollect devices and AveloMasks were received for use in the study.
For downstream analysis, the team agreed to proceed with nucleic acid extraction using the QIAsymphony platform, and finalisation of the analytical workflow is happening based on the data generated from WP2. Recruitment for the clinical study is due to commence in March 2025 to co-incide with the South African respiratory viral season.
Objective 3: Evaluate the diagnostic accuracy of XBA sampling in symptomatic adults, and feasibility of XBA sampling for detecting asymptomatic infection
The project’s clinical studies will be conducted under Objective 3, whereby Project B-Path aims to evaluate the diagnostic accuracy of an exhaled breath aerosol sample in symptomatic adults, and feasibility of sampling for detecting asymptomatic infection. The multisite coordinated study protocol has been developed to be implemented in Bucharest (Romania), Milan (Italy) and Heidelberg (Germany). Ethical approvals have been received, the trial registered in the German Clinical Trial Registry and the studies were registered as the different teams prepared each site for recruitment commencement in March 2026.
Objective 4: Evaluate acceptability, usability, population impact, and cost-effectiveness of XBA sampling for diagnosis and screening of respiratory infections of pandemic potential
Initial work on human factors engineering has been completed with some valuable insights gathered around the mask design, packaging, hygiene, acceptability and consideration for adaptation. A mixed-methods study of 22 participants with both AveloCollect and AveloMask lay the groundwork for evaluating the devices during the remainder of the project. Plans to further assess acceptability, appropriateness and usability are nested within the study protocols under Objective 3 and will take place in Germany, Romania and Italy in adults being diagnosed and screened for respiratory infections.
Population impact and cost-effectiveness of XBA will be analysed in the second half of the project.
To date, direct-pathogen detection from XBA using a commercial portable sampling device has not been possible. Avelo and consortium partners have optimised workflows for multiplex molecular detection of respiratory pathogens from breath samples. Further results from clinical studies, qualitative and modelling data will become available in the subsequent parts of the project.