Useful molecular tools for facing next pandemic events: Effective sample preparation and improved RT-PCR for highly sensitive detection of SARS-CoV-2 in wastewater environment

Rusková, Magdaléna; Bučková, Maria; Achs, Adam; Puškárová, Andrea; Wu, Jer Horng; Kuchta, T.; Šubr, Z; Pangallo, Domenico

doi:10.1016/j.ijheh.2022.114017

Cited by 3 publications

(9 citation statements)

References 33 publications

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“…Single-tube nested PCR allows the reaction to be performed in a single PCR tube, reducing the possibility of cross-contamination. The detection performance of the single-tube one-step nested quantitative PCR (OSN-qRT-PCR) method and the RT-qPCR method was evaluated by Rusková et al ( 2022 ). The LOD of OSN-qRT-PCR was the first order of magnitude higher than that of RT-qPCR and was able to improve virus detection, indicating that OSN-qRT-PCR can sensitively detect SARS-CoV-2 in a wastewater environment (Rusková et al 2022 ).…”

Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 surveillance in medical and industrial wastewater—a global perspective: a narrative review

Yang

Guo

Møhlenberg

et al. 2023

Environ Sci Pollut Res

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The novel coronavirus SARS-CoV-2 has spread at an unprecedented rate since late 2019, leading to the global COVID-19 pandemic. During the pandemic, being able to detect SARS-CoV-2 in human populations with high coverage quickly is a huge challenge. As SARS-CoV-2 is excreted in human excreta and thus exposed to the aqueous environment through sewers, the goal is to develop an ideal, non-invasive, cost-effective epidemiological method for detecting SARS-CoV-2. Wastewater surveillance has gained widespread interest and is increasingly being investigated as an effective early warning tool for monitoring the spread and evolution of the virus. This review emphasizes important findings on SARS-CoV-2 wastewater-based epidemiology (WBE) in different continents and techniques used to detect SARS-CoV-2 in wastewater during the period 2020–2022. The results show that WBE is a valuable population-level method for monitoring SARS-CoV-2 and is a valuable early warning alert. It can assist policymakers in formulating relevant policies to avoid the negative impacts of early or delayed action. Such strategy can also help avoid unnecessary wastage of medical resources, rationalize vaccine distribution, assist early detection, and contain large-scale outbreaks.

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Section: Resultsmentioning

confidence: 99%

SARS-CoV-2 surveillance in medical and industrial wastewater—a global perspective: a narrative review

Yang

Guo

Møhlenberg

et al. 2023

Environ Sci Pollut Res

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“…Several sets of primers and probes were designed using the software Primer3Plus ( https://dev.primer3plus.com/index.html ). The first group of primers and probes was oriented to the SARS-CoV-2 spike protein gene ( S ) (Rusková et al 2022 ). The second group of primers and probes was oriented to nucleocapsid phosphoprotein gene ( N ; accession number: MZ054877).…”

Section: Methodsmentioning

confidence: 99%

Comparison of ordinary reverse transcription real-time polymerase chain reaction (qRT-PCR) with a newly developed one-step single-tube nested real-time RT-PCR (OSN-qRT-PCR) for sensitive detection of SARS-CoV-2 in wastewater

Rusková

Bučková

Puškárová

et al. 2023

Environ Sci Pollut Res

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Wastewater monitoring has proven to be an important approach to detecting and controlling the development of the SARS-CoV-2 pandemic. Various tests based on reverse transcription real-time PCR (qRT-PCR) have been developed and used for the detection of SARS-CoV-2 in wastewater samples. In this study, we attempted to increase the sensitivity of qRT-PCR by developing a one-step single-tube nested qRT-PCR assay (OSN-qRT-PCR). Two variants were developed, oriented to nucleocapsid phosphoprotein gene (N) and to spike protein gene (S), respectively. The performance of conventional qRT-PCR assays oriented to these genes with two novel OSN-qRT-PCR assays were firstly optimized using wastewater artificially contaminated with two encapsidated RNA mimic systems harboring a portion either N or S gene (ENRM and ESRM, respectively). The assays were coupled to a polyethylene glycol–based RNA precipitation/extraction method and applied to detect SARS-CoV-2 in wastewater samples from four cities in Slovakia. Both novel OSN-qRT-PCR assays demonstrated higher detection rates than the ordinary qRT-PCR counterparts. The virus levels in the analyzed wastewater samples had a high or very high relation with the numbers of clinical cases in the monitored regions. In fact, correlation with a 3-, 4-, or 5-day temporal offset was revealed. The OSN-qRT-PCR assays demonstrated robustness, mainly in samples with low viral loads.

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Section: Computational Framework For Covid-19 Diagnostics and Therape...mentioning

confidence: 99%

“…RT-PCR is generally a couple-of-step technique for evaluating viral concentration from effective samples [ 102 ]. One-advance constant RT-PCR runs the entire RT-PCR reaction on a single cylinder that contains the essential preliminaries [ 102 ]. The two-advance continuous RT-PCR method uses multiple cylinders to conduct independent converse translation and intensification reactions.…”

Section: Computational Framework For Covid-19 Diagnostics and Therape...mentioning

confidence: 99%

“…The two-advance continuous RT-PCR method uses multiple cylinders to conduct independent converse translation and intensification reactions. Yet, it is more adaptable and effective than the one-advance method [ 102 ], fewer raw ingredients are needed, and cDNA can be stored for later objective examination. The one-advance technique is frequently chosen for SARS-CoV-2 detection because it is swiftly balanced, comprises limited example handling, reduces track time, lowers the risk of error and contamination, and continuously combines RT and PCR stages.…”

Section: Computational Framework For Covid-19 Diagnostics and Therape...mentioning

confidence: 99%

See 1 more Smart Citation

Reviewing methods of deep learning for diagnosing COVID-19, its variants and synergistic medicine combinations

Rafique,

Rehman,

Afghan

et al. 2023

Computers in Biology and Medicine

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Useful molecular tools for facing next pandemic events: Effective sample preparation and improved RT-PCR for highly sensitive detection of SARS-CoV-2 in wastewater environment

Cited by 3 publications

References 33 publications

SARS-CoV-2 surveillance in medical and industrial wastewater—a global perspective: a narrative review

SARS-CoV-2 surveillance in medical and industrial wastewater—a global perspective: a narrative review

Comparison of ordinary reverse transcription real-time polymerase chain reaction (qRT-PCR) with a newly developed one-step single-tube nested real-time RT-PCR (OSN-qRT-PCR) for sensitive detection of SARS-CoV-2 in wastewater

Reviewing methods of deep learning for diagnosing COVID-19, its variants and synergistic medicine combinations

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