Using Molecular Tools to Compare Various Concentration Protocols Prior to Detecting SARS-Cov-2 RNA in Wastewater

Document Type : Original Article


1 Water pollution research department, institute of environmental research and climate change, National research center,giza,Egypt

2 Therapeutic chemistry department, Institute of pharmaceutical and drug industry research, National research center, Giza, Egypt

3 Therapeutic Chemistry Department, National Research Centre (NRC), 33 EL Bohouth St. (former El Tahrir St.),


Background: Reports showed presence of SARS-CoV-2 genetic material in wastewater. Wastewater concentration methods are optimized for detection of non-enveloped viruses so need to be adopted for enveloped viruses and their genetic material. Methods: Conventional (cRT-PCR) and quantitative real time RT-PCR (qRT-PCR) were used as readouts to compare 4 water concentration methods namely, A) filtration on negatively charged membrane followed by extracting RNA from it, B) adsorbtion-elution method, C) flocculation with skimmed milk and D) polyethylene glycol precipitation, to detect SARS-CoV-2 RNA and 229E human coronavirus (229E-HCoV) as a model for spike-containing enveloped virus from fresh and wastewater. Results: On using cRT-PCR: recovery rate of SARS-CoV-2 RNA was better using method A then B for fresh water and method B then D for wastewater. 229E-HCoV recovery from fresh water was better using method C then A and methods B then D for wastewater. On using qRT-PCR, both methods A and B were better for SARS-CoV-2 RNA recovery from both fresh and wastewater. For the 229E-HCoV methods A was the most efficient for fresh water and method B for wastewater. Conclusion: Method B is recommended for SARS-CoV-2 RNA or whole 229E-HCoV recovery from wastewater.


Main Subjects

Volume 65, Issue 131 - Serial Number 13
Special Issue: Chemistry and Global Challenges (Part A)
December 2022
Pages 1241-1248
  • Receive Date: 25 September 2022
  • Revise Date: 22 November 2022
  • Accept Date: 11 December 2022