An efficient method to enhance recovery and detection of SARS-CoV-2 RNA in wastewater

Kumblathan, Teresa; Liu, Yanming; Qiu, Yuanyuan; Pang, Lilly; Hrudey, Steve E.; Le, X. Chris; Li, Xing‐Fang

doi:10.1016/j.jes.2022.10.006

Cited by 21 publications

(17 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Diverse sampling and analytical methods were identified as contributing factors for low accuracy and reproducibility for virus quantification in wastewater [ 17 ]. Current analytical approaches recover about 10–80% of the SARS-CoV-2 RNA from wastewater [ 17 , 28 , 29 ]. It is thus critical to improve the reproducibility and comparability of results by standardizing wastewater sampling, analysis, and data reporting protocols.…”

Section: Future Research To Improve the Wbe Methodologiesmentioning

confidence: 99%

Moving forward with COVID-19: Future research prospects of wastewater-based epidemiology methodologies and applications

Jiang

Liu

Tang

et al. 2023

Current Opinion in Environmental Science & Health

View full text Add to dashboard Cite

Section: Future Research To Improve the Wbe Methodologiesmentioning

confidence: 99%

Moving forward with COVID-19: Future research prospects of wastewater-based epidemiology methodologies and applications

Jiang

Liu

Tang

et al. 2023

Current Opinion in Environmental Science & Health

View full text Add to dashboard Cite

“…Both the liquid and solid phases of wastewater sewage samples were processed to capture viral materials for the subsequent analysis. 21 An 80 mL aliquot of each wastewater sewage sample was centrifuged at 3000 rpm for 30 min. The resulting supernatant was transferred into a separate tube.…”

Section: Methodsmentioning

confidence: 99%

Multiplex Assays Enable Simultaneous Detection and Identification of SARS-CoV-2 Variants of Concern in Clinical and Wastewater Samples

et al. 2023

Self Cite

View full text Add to dashboard Cite

The targeted screening and sequencing approaches for COVID-19 surveillance need to be adjusted to fit the evolving surveillance objectives which necessarily change over time. We present the development of variant screening assays that can be applied to new targets in a timely manner and enable multiplexing of targets for efficient implementation in the laboratory. By targeting the HV69/70 deletion for Alpha, K417N for Beta, K417T for Gamma, and HV69/ 70 deletion plus K417N for sub-variants BA.1, BA.3, BA.4, and BA.5 of Omicron, we achieved simultaneous detection and differentiation of Alpha, Beta, Gamma, and Omicron in a single assay. Targeting both T478K and P681R mutations enabled specific detection of the Delta variant. The multiplex assays used in combination, targeting K417N and T478K, specifically detected the Omicron sub-variant BA.2. The limits of detection for the five variants of concern were 4−16 copies of the viral RNA per reaction. Both assays achieved 100% clinical sensitivity and 100% specificity. Analyses of 377 clinical samples and 24 wastewater samples revealed the Delta variant in 100 clinical samples (nasopharyngeal and throat swab) collected in November 2021. Omicron BA.1 was detected in 79 nasopharyngeal swab samples collected in January 2022. Alpha, Beta, and Gamma variants were detected in 24 wastewater samples collected in May−June 2021 from two major cities of Alberta (Canada), and the results were consistent with the clinical cases of multiple variants reported in the community.

show abstract

“…WS can provide a realistic scale of the pandemic because it can capture input from all the symptomatic, presymptomatic, and asymptomatic individuals in a whole community served by a common sewer system. This alternative approach for assessing the status of overall community infection of SARS-CoV-2 has been supported by various organizations such as the U.S. Centers for Disease Control and Prevention, the International Water Association, the Global Institute for Water Security, the Public Health Agency of Canada, and the Canadian Water Network. − It is worth noting that our group has developed an efficient WS protocol with an 80% recovery rate, which is much higher than the recovery rates reported by other studies. − This efficient WS protocol provides excellent detection sensitivity of SARS-CoV-2 RNA in wastewater …”

Section: Introductionmentioning

confidence: 95%

Quantification and Differentiation of SARS-CoV-2 Variants in Wastewater for Surveillance

et al. 2023

Self Cite

View full text Add to dashboard Cite

Wastewater surveillance plays an important role in the monitoring of infections of SARS-CoV-2 at the community level. We report here the determination of SARS-CoV-2 and differentiation of its variants of concern in 294 wastewater samples collected from two major Canadian cities from May 2021 to March 2023. The overall method of analysis involved extraction of the virus and viral components using electronegative membranes, in situ stabilization and concentration of the viral RNA onto magnetic beads, and direct analysis of the viral RNA on the magnetic beads. Multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays, targeting specific and naturally selected mutations in SARS-CoV-2, enabled detection and differentiation of the Alpha, Beta, Gamma, Delta, and Omicron variants. An Omicron triplex RT-qPCR assay targeting three mutations, HV 69−70 deletion, K417N, and L452R, was able to detect and differentiate the Omicron BA.1/BA.3, BA.2/XBB, and BA.4/5. This assay had efficiencies of 90−104% for all three mutation targets and a limit of detection of 28 RNA copies per reaction. Analyses of 294 wastewater samples collected over a two-year span showed the concentrations and trends of Alpha, Beta, Gamma, Delta, and Omicron variants as they emerge in two major Canadian cities participating in the wastewater surveillance program. The trends of specific variants were consistent with clinical reports for the same period. At the beginning of each wave, the corresponding variants were detectable in wastewater. For example, RNA concentrations of the BA.2 variant were as high as 10 4 copies per 100 mL of wastewater collected in January 2022, when approximately only 50−60 clinical cases of BA.2 infection were reported in Canada. These results show that the strategy and highly sensitive assays for the variants of concern in wastewater are potentially useful for the detection of newly emerging SARS-CoV-2 variants and other viruses for future community biomonitoring.

show abstract

An efficient method to enhance recovery and detection of SARS-CoV-2 RNA in wastewater

Cited by 21 publications

References 55 publications

Moving forward with COVID-19: Future research prospects of wastewater-based epidemiology methodologies and applications

Moving forward with COVID-19: Future research prospects of wastewater-based epidemiology methodologies and applications

Multiplex Assays Enable Simultaneous Detection and Identification of SARS-CoV-2 Variants of Concern in Clinical and Wastewater Samples

Quantification and Differentiation of SARS-CoV-2 Variants in Wastewater for Surveillance

Contact Info

Product

Resources

About