This protocol describes materials and methods that can be employed in the field to collect Moore ... more This protocol describes materials and methods that can be employed in the field to collect Moore swabs and/or grab samples for wastewater sample collection. The materials and methods described herein are meant to provide guidelines, examples, and tips for various methods of wastewater collection; however, these techniques can be adapted and modified depending on the setting and goals of the particular project or program. This protocol makes note of composite sampling instructions but is focused on situations where installation and operation of a composite sampler is not feasible.
This protocol details a method for SARS-CoV-2 capture and concentration through the use of Nanotr... more This protocol details a method for SARS-CoV-2 capture and concentration through the use of Nanotrap® Magnetic Virus Particles from 40mL of wastewater sample.
As COVID-19 continues to spread globally, monitoring the disease at different scales is critical ... more As COVID-19 continues to spread globally, monitoring the disease at different scales is critical to support public health decision making. Surveillance for SARS-CoV-2 RNA in wastewater can supplement surveillance based on diagnostic testing. In this paper, we report the results of wastewater-based COVID-19 surveillance on Emory University campus that included routine sampling of sewage from a hospital building, an isolation/quarantine building, and 21 student residence halls between July 13th, 2020 and March 14th, 2021. We examined the sensitivity of wastewater surveillance for detecting COVID-19 cases at building level and the relation between Ct values from RT-qPCR results of wastewater samples and the number of COVID-19 patients residing in the building. Our results show that weekly wastewater surveillance using Moore swab samples was not sensitive enough (6 of 63 times) to reliably detect one or two sporadic cases in a residence building. The Ct values of the wastewater samples over time from the same sampling location reflected the temporal trend in the number of COVID-19 patients in the isolation/quarantine building and hospital (Pearson's r < −0.8), but there is too much uncertainty to directly estimate the number of COVID-19 cases using Ct values. After students returned for the spring 2021 semester, SARS-CoV-2 RNA was detected in the wastewater samples from most of the student residence hall monitoring sites one to two weeks before COVID-19 cases surged on campus. This finding suggests that wastewater-based surveillance can be used to provide early warning of COVID-19 outbreaks at institutions.
This protocols describes the use of Skim Milk Flocculation to concentrate and extract SARS-CoV-2 ... more This protocols describes the use of Skim Milk Flocculation to concentrate and extract SARS-CoV-2 from Moore swab wastewater samples. Included in this protocol are: materials and equipment, steps for processing Moore swabs, and steps for concentrating and extracting the virus using the Qiagen RNeasy Mini Kit.
These protocols have been adapted from Ceres "Nanotrap® Wastewater Protocol using MagMAX Kit... more These protocols have been adapted from Ceres "Nanotrap® Wastewater Protocol using MagMAX Kits" (APP-030, Revision 0, Nov 2021). This protocol uses Nanotrap® Magnetic Virus Particles and Nanotrap® Enhancement Reagent 1 (ER1) to capture and concentrate viruses in wastewater samples. It is optimized for viral capture from 10 mL samples of wastewater and is compatible with three nucleic acid extraction kits from ThermoFisher. This protocol has been used and adapted for SARS-CoV-2 viral capture at Emory University for wastewater surveillance of COVID-19.
SARS-CoV-2 is a respiratory virus, but it is also detected in a significant proportion of fecal s... more SARS-CoV-2 is a respiratory virus, but it is also detected in a significant proportion of fecal samples from COVID-19 cases. Recent studies have shown that wastewater surveillance can be a low-cost tool compared to massive diagnostic testing for tracking COVID-19 outbreaks in communities, but most studies have focused on sampling from wastewater treatment plants. Institutional level wastewater surveillance may serve well for early warning purposes because specific geographic areas/populations with emerging cases can be tracked and immediate action can be executed in the event of a positive wastewater signal. In this study, a novel Moore swab method was developed and used for wastewater surveillance of COVID-19 at an institutional level. Of the 442 swab samples tested, 148 (33.5%) swabs collected from the three campuses and two buildings were positive for SARS-CoV-2 RNA. Further study of the quarantine building with a known number of cases indicated that this method was sensitive enough to detect few cases in the building. In addition, comparison between grab samples and Moore swab samples from the hospital sewage line indicated that Moore swabs were more sensitive than grab samples and offer a simple, inexpensive method for obtaining a composite sample of virus in wastewater over a 24–48 h period. These results suggest that collection and analyses of Moore swabs for SARS-CoV-2 RNA detection is a sensitive, low-cost, and easy to use tool for COVID-19 surveillance that is useful for institutional settings and could be deployed in low-resource settings to identify emerging COVID-19 clusters in communities.
ABSTRACTSARS-CoV-2 is a respiratory virus but it is also detected in a significant proportion of ... more ABSTRACTSARS-CoV-2 is a respiratory virus but it is also detected in a significant proportion of fecal samples of COVID-19 cases. Recent studies have shown that wastewater surveillance can be a low-cost tool for management of COVID-19 pandemic and tracking COVID-19 outbreaks in communities but most studies have been focusing on sampling from wastewater treatment plants. Institutional level of wastewater surveillance may serve well for early warning purposes since cases can be tracked and immediate action can be executed in the event of positive signal. In this study, a novel Moore swab method was developed and used for wastewater surveillance of COVID-19 at institutional level. Among the 219 swab samples tested, 28 (12.8%) swabs collected from the three campuses and two buildings were positive for SARS-CoV-2. Further individual clinical diagnosis validated the wastewater results and indicated that this method was sensitive enough to detect 1-2 cases in a building. In addition, compa...
Monitoring SARS-CoV-2 in wastewater is a valuable approach to track COVID-19 transmission. Design... more Monitoring SARS-CoV-2 in wastewater is a valuable approach to track COVID-19 transmission. Designing wastewater surveillance (WWS) with representative sampling sites and quantifiable results requires knowledge of the sewerage system and virus fate and transport. We developed a multi-level WWS system to track COVID-19 in Atlanta using an adaptive nested sampling strategy. From March 2021 to April 2022, 868 wastewater samples were collected from influent lines to wastewater treatment facilities and upstream community manholes. Variations in SARS-CoV-2 concentrations in influent line samples preceded similar variations in numbers of reported COVID-19 cases in the corresponding catchment areas. Community sites under nested sampling represented mutually-exclusive catchment areas. Community sites with high SARS-CoV-2 detection rates in wastewater covered high COVID-19 incidence areas, and adaptive sampling enabled identification and tracing of COVID-19 hotspots. This study demonstrates ho...
This protocol describes materials and methods that can be employed in the field to collect Moore ... more This protocol describes materials and methods that can be employed in the field to collect Moore swabs and/or grab samples for wastewater sample collection. The materials and methods described herein are meant to provide guidelines, examples, and tips for various methods of wastewater collection; however, these techniques can be adapted and modified depending on the setting and goals of the particular project or program. This protocol makes note of composite sampling instructions but is focused on situations where installation and operation of a composite sampler is not feasible.
This protocol details a method for SARS-CoV-2 capture and concentration through the use of Nanotr... more This protocol details a method for SARS-CoV-2 capture and concentration through the use of Nanotrap® Magnetic Virus Particles from 40mL of wastewater sample.
As COVID-19 continues to spread globally, monitoring the disease at different scales is critical ... more As COVID-19 continues to spread globally, monitoring the disease at different scales is critical to support public health decision making. Surveillance for SARS-CoV-2 RNA in wastewater can supplement surveillance based on diagnostic testing. In this paper, we report the results of wastewater-based COVID-19 surveillance on Emory University campus that included routine sampling of sewage from a hospital building, an isolation/quarantine building, and 21 student residence halls between July 13th, 2020 and March 14th, 2021. We examined the sensitivity of wastewater surveillance for detecting COVID-19 cases at building level and the relation between Ct values from RT-qPCR results of wastewater samples and the number of COVID-19 patients residing in the building. Our results show that weekly wastewater surveillance using Moore swab samples was not sensitive enough (6 of 63 times) to reliably detect one or two sporadic cases in a residence building. The Ct values of the wastewater samples over time from the same sampling location reflected the temporal trend in the number of COVID-19 patients in the isolation/quarantine building and hospital (Pearson's r < −0.8), but there is too much uncertainty to directly estimate the number of COVID-19 cases using Ct values. After students returned for the spring 2021 semester, SARS-CoV-2 RNA was detected in the wastewater samples from most of the student residence hall monitoring sites one to two weeks before COVID-19 cases surged on campus. This finding suggests that wastewater-based surveillance can be used to provide early warning of COVID-19 outbreaks at institutions.
This protocols describes the use of Skim Milk Flocculation to concentrate and extract SARS-CoV-2 ... more This protocols describes the use of Skim Milk Flocculation to concentrate and extract SARS-CoV-2 from Moore swab wastewater samples. Included in this protocol are: materials and equipment, steps for processing Moore swabs, and steps for concentrating and extracting the virus using the Qiagen RNeasy Mini Kit.
These protocols have been adapted from Ceres "Nanotrap® Wastewater Protocol using MagMAX Kit... more These protocols have been adapted from Ceres "Nanotrap® Wastewater Protocol using MagMAX Kits" (APP-030, Revision 0, Nov 2021). This protocol uses Nanotrap® Magnetic Virus Particles and Nanotrap® Enhancement Reagent 1 (ER1) to capture and concentrate viruses in wastewater samples. It is optimized for viral capture from 10 mL samples of wastewater and is compatible with three nucleic acid extraction kits from ThermoFisher. This protocol has been used and adapted for SARS-CoV-2 viral capture at Emory University for wastewater surveillance of COVID-19.
SARS-CoV-2 is a respiratory virus, but it is also detected in a significant proportion of fecal s... more SARS-CoV-2 is a respiratory virus, but it is also detected in a significant proportion of fecal samples from COVID-19 cases. Recent studies have shown that wastewater surveillance can be a low-cost tool compared to massive diagnostic testing for tracking COVID-19 outbreaks in communities, but most studies have focused on sampling from wastewater treatment plants. Institutional level wastewater surveillance may serve well for early warning purposes because specific geographic areas/populations with emerging cases can be tracked and immediate action can be executed in the event of a positive wastewater signal. In this study, a novel Moore swab method was developed and used for wastewater surveillance of COVID-19 at an institutional level. Of the 442 swab samples tested, 148 (33.5%) swabs collected from the three campuses and two buildings were positive for SARS-CoV-2 RNA. Further study of the quarantine building with a known number of cases indicated that this method was sensitive enough to detect few cases in the building. In addition, comparison between grab samples and Moore swab samples from the hospital sewage line indicated that Moore swabs were more sensitive than grab samples and offer a simple, inexpensive method for obtaining a composite sample of virus in wastewater over a 24–48 h period. These results suggest that collection and analyses of Moore swabs for SARS-CoV-2 RNA detection is a sensitive, low-cost, and easy to use tool for COVID-19 surveillance that is useful for institutional settings and could be deployed in low-resource settings to identify emerging COVID-19 clusters in communities.
ABSTRACTSARS-CoV-2 is a respiratory virus but it is also detected in a significant proportion of ... more ABSTRACTSARS-CoV-2 is a respiratory virus but it is also detected in a significant proportion of fecal samples of COVID-19 cases. Recent studies have shown that wastewater surveillance can be a low-cost tool for management of COVID-19 pandemic and tracking COVID-19 outbreaks in communities but most studies have been focusing on sampling from wastewater treatment plants. Institutional level of wastewater surveillance may serve well for early warning purposes since cases can be tracked and immediate action can be executed in the event of positive signal. In this study, a novel Moore swab method was developed and used for wastewater surveillance of COVID-19 at institutional level. Among the 219 swab samples tested, 28 (12.8%) swabs collected from the three campuses and two buildings were positive for SARS-CoV-2. Further individual clinical diagnosis validated the wastewater results and indicated that this method was sensitive enough to detect 1-2 cases in a building. In addition, compa...
Monitoring SARS-CoV-2 in wastewater is a valuable approach to track COVID-19 transmission. Design... more Monitoring SARS-CoV-2 in wastewater is a valuable approach to track COVID-19 transmission. Designing wastewater surveillance (WWS) with representative sampling sites and quantifiable results requires knowledge of the sewerage system and virus fate and transport. We developed a multi-level WWS system to track COVID-19 in Atlanta using an adaptive nested sampling strategy. From March 2021 to April 2022, 868 wastewater samples were collected from influent lines to wastewater treatment facilities and upstream community manholes. Variations in SARS-CoV-2 concentrations in influent line samples preceded similar variations in numbers of reported COVID-19 cases in the corresponding catchment areas. Community sites under nested sampling represented mutually-exclusive catchment areas. Community sites with high SARS-CoV-2 detection rates in wastewater covered high COVID-19 incidence areas, and adaptive sampling enabled identification and tracing of COVID-19 hotspots. This study demonstrates ho...
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