Lab Matters Fall 2023 | Page 111

APHL 2023 POSTER ABSTRACTS

Going With the Flow : Implementing a COVID-19 Wastewater Surveillance Program in a Small State

C . Benton , State of New Hampshire Public Health Laboratories

The State of New Hampshire Public Health Laboratory ( NH PHL ) implemented wastewater testing for COVID-19 to act as an early outbreak warning system for the state ’ s 1.5 million residents . Implementation of this testing required overcoming several challenges , such as identifying laboratory space , personnel , and equipment . Cross training of staff , utilizing shared lab space , and redirecting laboratory funds as well as partnering with the University of New Hampshire enabled NH PHL to efficiently bring on this testing . As UNH had an established wastewater surveillance program on their campus , NH PHL was able to share protocols , and received guidance for implementation . Wastewater surveillance for COVID-19 began on October 1 , 2022 , and currently consists of 14 sewage treatment plants throughout the state submitting weekly samples for testing . The submissions from each facility are effluent collected over a 24 hour period . Once received at the lab , the samples are processed and extracted using the KingFisher Apex platform , and digital PCR for COVID-19 detection is performed using the Qiagen Qiacuity . The data that is collected is then used to populate a publicly facing COVID-19 dashboard , allowing the general public to view trends in COVID-19 wastewater by facility . Currently , New Hampshire is expanding this testing to include WGS of these samples for variant determination , and is also working to add testing for additional pathogens .

Presenter : Christopher Benton , christopher . benton @ dhhs . nh . gov

Lessons Learned from Two Years of SARS-CoV-2 Wastewater Surveillance : Comparison of Viral Concentration , Extraction and Quantification Methods

D . Antkiewicz 1 , 2 , K . Janssen 1 , A . Roguet 1 , H . Pilch 2 , R . Fahney 1 , P . Mullen 1 , D . Everett 1 , G . Knuth 1 , E . Doolittle 1 , A . Stanley 1 , 2 , J . Hemming 1 , M . Shafer 1 , 2 ; 1 Wisconsin State Laboratory of Hygiene ,

2

University of Wisconsin Madison

Wastewater monitoring of SARS-CoV-2 throughout the COVID-19 pandemic has documented the utility of the wastewater-based epidemiology ( WBE ) approach as a cost-effective , fast and relatively unbiased method for assessing trends in community-wide viral concentrations . Just two years since the inception of many of the SARS-CoV-2 WBE programs there are now over 3,800 sites across 70 countries and over 150 public facing dashboards devoted to WBE surveillance . The pressing need for wastewater-specific viral methods motivated the rapid development of a large number of protocols and tools and that effort continues to date . Thus far , the WBE community gained a deeper understanding of the advantages and limitations of some of the protocols . Still , even though new guidelines have been written both by the CDC and the National Academy of Sciences , the information on which to judge the comparative performance of the methods for concentration , extraction and quantification of SARS-CoV-2 from wastewater remains limited . Despite the rapid evolution of WBE methods and an impressive body of literature emerging from those efforts , much remains to be evaluated and standardized . Our work attempts to systematically compare the most popular WBE methods in a stepby-step approach , including comparisons of HA electronegative filtration to Nanotrap ® Microbiome A particles and polyethylene glycol ( PEG ) precipitation , as well as the direct capture using

Promega ’ s PureYield binding column . We also evaluated different extraction methods and compared side-by-side qPCR with digital PCR platform . Overall , our data suggest that multiple concentration methods can perform adequately , though some stark differences can be observed with specific wastewater matrices . Among the methods tested , the automated paramagnetic bead-based extraction yielded better viral recoveries than the manual on-column extraction . In addition , the viral target quantification with traditional qPCR was found to perform adequately when careful quality control was applied , but the digital PCR was more sensitive and slightly less variable . In particular , the combination of Nanotrap ® concentration with automated bead-based extraction and digital PCR quantification was chosen as the most practical and best performing WBE method .

Presenter : Dagmara Antkiewicz , dksieprawska @ wisc . edu

SARS-CoV-2 Wastewater Workflow Comparison for Upscale or Downscale Testing in the National Capital Region ( NCR ) Composed of the District of Columbia and Southern Maryland

E . Zeleya , M . Mann , A . Park , H . Tesfagebriel , D . Payne , J . Hauser , S . Byfield ; District of Columbia Department of Forensic Sciences , Public Health Laboratory Division

Clinical monitoring of the COVID-19 pandemic has been essential , but unintentionally taxing on the public ’ s willingness to undergo nasopharyngeal swabbing or administer at-home self-tests . Population-level surveillance can be performed to supplement individual clinical tests . Infected persons shed SARS-CoV-2 in their feces , urine , sweat , and respiratory fluids . Research by CDC and WHO have demonstrated a correlation between SARS-CoV-2 found in wastewater and the SARS-CoV-2 infection rate found in clinical testing . By surveilling raw wastewater within the National Capital Region ( NCR ), the District of Columbia Public Health Laboratory ( DC PHL ) is able to surveil all eight wards of the District of Colombia , various K-12 schools during the academic year and parts of southern Maryland . For processing and analysis of wastewater samples , we first concentrate the raw wastewater , extract viral RNA , quantify SARS-CoV-2 abundance using digital PCR , sequence variant strains using the Illumina MiSeq , then transfer data to local and federal partners . The DC PHL utilizes one of two methods for the initial concentration step : the Innova Prep Concentrating Pipette or the Ceres Nanotrap Technology . The Innova Prep method concentrates one wastewater sample at a time in less than five minutes by capturing viral particles in the pores of an ultra-filtrating pipette tip . Whereas Ceres Nanotrap is a high throughput system that can concentrate 24 wastewater samples in 40 minutes and achieves viral capture through high affinity rapid binding . The purpose of this study is to compare these two methods based on utilization time , throughput , cost and how suitable the products are for downstream applications , including sequencing and digital PCR using the QIAcuity Digital PCR Instrument . This study will provide data to aid public health laboratories in optimizing workflows for analyzing wastewater samples for SARS-CoV-2 , but also for other pathogens found in wastewater such as mpox , Candida auris and various other respiratory diseases .

Presenter : Shauntelle Byfield , shauntelle . byfield @ dc . gov