Building-level wastewater surveillance using tampon swabs and RT-LAMP for rapid SARS-CoV-2 RNA detection

Bivins, Aaron; Lott, M.; Shaffer, Marlee; Wu, Zhenyu; North, Devin; Lipp, Erin K.; Bibby, Kyle

doi:10.1039/d1ew00496d

Cited by 42 publications

(41 citation statements)

References 52 publications

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“… 6 − 17 This approach uses a series of complex environmental microbiology procedures ranging from wastewater sampling to the detection and quantification of viral RNA primarily via polymerase chain reaction (PCR)-based or loop-mediated isothermal amplification (LAMP) assays. 6 , 8 , 18 These procedures require careful optimization to maximize their sensitivity for detecting traces of SARS-CoV-2 in wastewater, especially in the absence of standard methods. 19 − 21 Trace detection, the reliable detection of minuscule amounts of RNA in wastewater, is crucial for regions with few COVID-19 cases and for jurisdictions in which elimination is a management strategy.…”

Section: Introductionmentioning

confidence: 99%

Comparison of RT-qPCR and RT-dPCR Platforms for the Trace Detection of SARS-CoV-2 RNA in Wastewater

et al. 2022

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We compared reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and RT digital PCR (RT-dPCR) platforms for the trace detection of SARS-CoV-2 RNA in low-prevalence COVID-19 locations in Queensland, Australia, using CDC N1 and CDC N2 assays. The assay limit of detection (ALOD), PCR inhibition rates, and performance characteristics of each assay, along with the positivity rates with the RT-qPCR and RT-dPCR platforms, were evaluated by seeding known concentrations of exogenous SARS-CoV-2 in wastewater. The ALODs using RT-dPCR were approximately 2–5 times lower than those using RT-qPCR. During sample processing, the endogenous ( n = 96) and exogenous ( n = 24) SARS-CoV-2 wastewater samples were separated, and RNA was extracted from both wastewater eluates and pellets (solids). The RT-dPCR platform demonstrated a detection rate significantly greater than that of RT-qPCR for the CDC N1 and CDC N2 assays in the eluate (N1, p = 0.0029; N2, p = 0.0003) and pellet (N1, p = 0.0015; N2, p = 0.0067) samples. The positivity results also indicated that for the analysis of SARS-CoV-2 RNA in wastewater, including the eluate and pellet samples may further increase the detection sensitivity using RT-dPCR.

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Section: Introductionmentioning

confidence: 99%

Comparison of RT-qPCR and RT-dPCR Platforms for the Trace Detection of SARS-CoV-2 RNA in Wastewater

et al. 2022

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“… 6 Bivins et al in 2021 employed tampons for passive sampling of SARS-CoV-2 RNA in wastewater collection systems. 9 Hayes et al in 2021 designed a 3D-printed passive sampling device that housed cotton cheesecloth or an electronegative filter, both of which were capable of passively adsorbing SARS-CoV-2 RNA in laboratory-controlled experiments and municipal sewersheds. 10 Of the two adsorbent materials evaluated in Hayes et al, electronegative filters were the preferred adsorbent material for passively capturing SARS-CoV-2 RNA in wastewater, as the laboratory-grade filters provide reproducibility and consistency.…”

Section: Introductionmentioning

confidence: 99%

Operational Constraints of Detecting SARS-CoV-2 on Passive Samplers using Electronegative Filters: A Kinetic and Equilibrium Analysis

et al. 2022

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In developing an effective monitoring program for the wastewater surveillance of SARS-CoV-2 ribonucleic acid (RNA), the importance of sampling methodology is paramount. Passive sampling has been shown to be an effective tool to detect SARS-CoV-2 RNA in wastewater. However, the adsorption characteristics of SARS-CoV-2 RNA on passive sampling material are not well-understood, which further obscures the relationship between wastewater surveillance and community infection. In this work, adsorption kinetics and equilibrium characteristics were evaluated using batch-adsorption experiments for heat-inactivated SARS-CoV-2 (HI-SCV-2) adsorption to electronegative filters. Equilibrium isotherms were assessed or a range of total suspended solids (TSS) concentrations (118, 265, and 497 mg L –1 ) in wastewater, and a modeled q max of 7 × 10 3 GU cm –2 was found. Surrogate adsorption kinetics followed a pseudo-first-order model in wastewater with maximum concentrations achieved within 24 h. In both field and isotherm experiments, equilibrium behavior and viral recovery were found to be associated with wastewater and eluate TSS. On the basis of the results of this study, we recommend a standard deployment duration of 24–48 h and the inclusion of eluate TSS measurement to assess the likelihood of solids inhibition during analysis.

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“…Within WBE, sample collection strategies vary with three predominant methodologies emerging: composite sampling, grab sampling and passive sampling. Autosamplers can mechanically pull wastewater samples routinely with defined time or flow intervals to create representative composite aliquots; however, these instruments are expensive and challenging to maintain in certain environments (Bivins et al, 2021). Grab samples are generally an operationally straightforward option, collecting a volume of wastewater at a single timepoint.…”

Section: Introductionmentioning

confidence: 99%

“…Grab samples are generally an operationally straightforward option, collecting a volume of wastewater at a single timepoint. However, such samples are susceptible to fecal shedding and flow fluctuations, particularly outside buildings with limited drainage areas, and may provide a biased sample of the wastewater stream (Bivins et al, 2021; Rafiee et al, 2021). Several universities have trialed the use of passive samplers in place of traditional composite or grab sampling methods due to additional cost-effectiveness and ease of deployment (Bivins et al, 2021; Brizee, 2021; Corchis-Scott et al, 2021; Severance, 2021).…”

Section: Introductionmentioning

confidence: 99%

“…However, such samples are susceptible to fecal shedding and flow fluctuations, particularly outside buildings with limited drainage areas, and may provide a biased sample of the wastewater stream (Bivins et al, 2021; Rafiee et al, 2021). Several universities have trialed the use of passive samplers in place of traditional composite or grab sampling methods due to additional cost-effectiveness and ease of deployment (Bivins et al, 2021; Brizee, 2021; Corchis-Scott et al, 2021; Severance, 2021). Schang et al demonstrated that the concentration of SARS-CoV-2 RNA recovered from passive samplers was positively correlated with the viral wastewater concentrations obtained using traditional sampling methods in communities with low caseloads (Schang et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

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Quantifying the relationship between SARS-CoV-2 wastewater concentrations and building-level COVID-19 prevalence at an isolation residence using a passive sampling approach

Acer

Kelly

Lover

et al. 2022

Preprint

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SARS-CoV-2 RNA can be detected in the excreta of individuals with COVID-19 and has demonstrated a positive correlation with various clinical parameters. Consequently, wastewater-based epidemiology (WBE) approaches have been implemented globally as a public health surveillance tool to monitor the community-level prevalence of infections. Over 270 higher education campuses monitor wastewater for SARS-CoV-2, with most gathering either composite samples via automatic samplers (autosamplers) or grab samples. However, autosamplers are expensive and challenging to manage with seasonal variability, while grab samples are particularly susceptible to temporal variation when sampling sewage directly from complex matrices outside residential buildings. Prior studies have demonstrated encouraging results utilizing passive sampling swabs. Such methods can offer affordable, practical, and scalable alternatives to traditional methods while maintaining a reproducible SARS-CoV-2 signal. In this regard, we deployed tampons as passive samplers outside of a COVID-19 isolation unit (a segregated residence hall) at a university campus from February 1, 2021 - May 21, 2021. Samples were collected several times weekly and remained within the sewer for a minimum of 24 hours (n = 64). SARS-CoV-2 RNA was quantified using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) targeting the viral N1 and N2 gene fragments. We quantified the mean viral load captured per individual and the association between the daily viral load and total persons, adjusting for covariates using multivariable models to provide a baseline estimate of viral shedding. Samples were processed through two distinct laboratory pipelines on campus, yielding highly correlated N2 concentrations. Data obtained here highlight the success of passive sampling utilizing tampons to capture SARS-CoV-2 in wastewater coming from a COVID-19 isolation residence, indicating that this method can help inform public health responses in a range of settings.

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Building-level wastewater surveillance using tampon swabs and RT-LAMP for rapid SARS-CoV-2 RNA detection

Abstract: Wastewater surveillance for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA has demonstrated useful correlation with both coronavirus disease 2019 (COVID-19) cases and clinical testing positivity at the community level....

Cited by 42 publications

References 52 publications

Comparison of RT-qPCR and RT-dPCR Platforms for the Trace Detection of SARS-CoV-2 RNA in Wastewater

Comparison of RT-qPCR and RT-dPCR Platforms for the Trace Detection of SARS-CoV-2 RNA in Wastewater

Operational Constraints of Detecting SARS-CoV-2 on Passive Samplers using Electronegative Filters: A Kinetic and Equilibrium Analysis

Quantifying the relationship between SARS-CoV-2 wastewater concentrations and building-level COVID-19 prevalence at an isolation residence using a passive sampling approach

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