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address how entities can participate in the Collaborative and explain what sampling and laboratory methods are required to ensure data consistency. Colorado Department of Public Health and Environment The Department will complete the following efforts: • Utilize the data to inform ongoing response to the COVID19 pandemic. • Provide technical assistance to Local Public Health Agencies on the data. • Work with Colorado State University, MSU and the Wastewater Utilities to optimize data collection and display of results. • Work with Colorado State University, MSU and the Wastewater Utilities to align wastewater collection catchment geographic information with COVID19 disease reporting information for correlation and display of results. • On a weekly basis, provide weekly surveillance data (case counts and positivity rates) aggregated by wastewater facility boundary or sewershed. This information will be used in the dashboard. • Will publish a link to the dashboard for Local Public Health Agencies and eventually the public. • Coordination with Centers for Disease Control with any national efforts to use wastewater surveillance. Colorado State University Colorado State University will complete the following: • Complete RNA extraction for wastewater influent samples. RNA will be extracted and concentrated via centrifugation, filtration and/or precipitation. • Viral RNA copy number will be quantified using reverse transcription PCR - either real-time PCR (for comparison to established procedures), or digital PCR (for increased sensitivity) • Data will be reported to the WWTP/State of Colorado as viral copies per liter. Trends in the data over time will also be reported. • Will lead development of and host the dashboard. Coordinate with the Department and the Wastewater Utilities on required GIS layers, information needed to calculate viral loads, and surveillance data to be displayed on the dashboard. Depending on length of surveillance effort the dashboard may eventually be transferred to the Department for ongoing updates and maintenance. • Laboratory capacity to process 38 samples/week (1,860 samples over course of 52 week period). Metropolitan State University of Denver MSU will complete the following: • MSU will work with the Department to identify specific questions associated with the statewide wastewater surveillance effort, and conduct experiments designed to answer these questions. MSU and the Department will get input on these questions from other members of the collaborative. The Department will prioritize the questions to align with the budget and MSU laboratory capacity. Examples of applicable projects might include questions about RNA stability, potential extraction techniques, and optimal sample processing prior to shipping. • Complete RNA extraction for wastewater influent samples. RNA will be extracted and concentrated via centrifugation, filtration and/or precipitation. • Viral RNA copy number will be quantified using reverse transcription PCR - real-time PCR (for comparison to established procedures) Page 3