Development of an integrated breath analysis technology for on-chip aerosol capture and molecular analysis

As proven early on in the pandemic, SARS-CoV-2 is mainly transmitted by aerosols. This urged us to develop a silicon impactor that collects the virus particles directly from breath. Performing PCR on these breath samples proved equally sensitive as nasopharyngeal swabs during the first week of an infection [Stakenborg et al., 2022], yet it remained a mostly manual process and PCR turn-around-time was still long. To overcome these drawbacks, we developed a fast and sensitive, fully integrated point-of-need breath test, comprising a novel breath sampler device and PCR instrument. The breath sampler combines virus collection and in-situ RNA amplification. The PCR instrument performs very fast amplification of the released viral RNA. Sample-to-result time was reduced to <20 min with an equal performance as the original manual procedure

Molecular detection of SARS-COV-2 in exhaled breath at the point-of-need

The SARS-CoV-2 pandemic has highlighted the need for improved technologies to help control the spread of contagious pathogens. While rapid point-of-need testing plays a key role in strategies to rapidly identify and isolate infectious patients, current test approaches have significant shortcomings related to assay limitations and sample type. Direct quantification of viral shedding in exhaled particles may offer a better rapid testing approach, since SARS-CoV-2 is believed to spread mainly by aerosols. It assesses contagiousness directly, the sample is easy and comfortable to obtain, sampling can be standardized, and the limited sample volume lends itself to a fast and sensitive analysis. In view of these benefits, we developed and tested an approach where exhaled particles are efficiently sampled using inertial impaction in a micromachined silicon chip, followed by an RT-qPCR molecular assay to detect SARS-CoV-2 shedding. Our portable, silicon impactor allowed for the efficient capture (>85%) of respiratory particles down to 300 nm without the need for additional equipment. We demonstrate using both conventional off-chip and in-situ PCR directly on the silicon chip that sampling subjects' breath in less than a minute yields sufficient viral RNA to detect infections as early as standard sampling methods. A longitudinal study revealed clear differences in the temporal dynamics of viral load for nasopharyngeal swab, saliva, breath, and antigen tests. Overall, after an infection, the breath-based test remains positive during the first week but is the first to consistently report a negative result, putatively signalling the end of contagiousness and further emphasizing the potential of this tool to help manage the spread of airborne respiratory infections