Graphene Multiplexed Sensor for Point-of-Need Viral Wastewater-Based Epidemiology

Wastewater-based epidemiology (WBE) can help mitigate the spread of respiratory infections through the early detection of viruses, pathogens, and other biomarkers in human waste. The need for sample collection, shipping, and testing facilities drives up the cost of WBE and hinders its use for rapid detection and isolation in environments with small populations and in low-resource settings. Given the ubiquitousness and regular outbreaks of respiratory syncytial virus, SARS-CoV-2 and various influenza strains, there is a rising need for a low-cost and easy-to-use biosensing platform to detect these viruses locally before outbreaks can occur and monitor their progression. To this end, we have developed an easy-to-use, cost-effective, multiplexed platform able to detect viral loads in wastewater with several orders of magnitude lower limit of detection than mass spectrometry. This is enabled by wafer scale production and aptamers pre-attached with linker molecules, producing forty-four chips at once. Each chip can simultaneously detect four target analytes using twenty transistors segregated into four sets of five for each analyte to allow for immediate statistical analysis. We show our platforms ability to rapidly detect three virus proteins (SARS-CoV-2, RSV, and Influenza A) and a population normalization molecule (caffeine) in wastewater. Going forward, turning these devices into hand-held systems would enable waste-water epidemiology in low-resource settings and be instrumental for rapid, local outbreak prevention. ### Competing Interest Statement Kenneth Stephen Burch reports financial support, equipment, drugs, or supplies, and writing assistance were provided by GRIP Molecular Inc. Kenneth Stephen Burch reports a relationship with GRIP Molecular Inc that includes: board membership and equity or stocks. Narendra Kumar reports a relationship with GRIP Molecular Inc that includes: equity or stocks. Kenneth Stephen Burch has patent pending to Boston College. Micheal Geiwitz has patent pending to Boston College. Stephen Hummel has patent pending to Boston College. Tim Van Opijnen has patent pending to Boston College. Narendra Kumar has patent pending to Boston College. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. ### Funding Statement NIH, Boston College Schiller Institute ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes Data available upon request.