Sewage surveillance for SARS-CoV-2: Molecular detection, quantification, and normalization factors.

The presence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in wastewater systems provides a primary indication of the coronavirus disease 2019 (COVID-19) spread throughout communities worldwide. Droplet digital polymerase chain reaction (dd-PCR) or reverse transcription-polymerase chain reaction (RT-PCR) administration of SARS-CoV-2 in wastewaters provides a reliable and efficient technology for gathering secondary local-level public health data. Often the accuracy of prevalence estimation is hampered by many methodological issues connected with wastewater surveillance. Still, more studies are needed to use and create efficient approaches for deciphering the actual SARS-CoV-2 indication from noise in the specimens/samples. Nearly 39-65% of positive patients and asymptomatic carriers expel the virus through their faeces however, only ∼6% of the infected hosts eject it through their urine. COVID-19 positive patients can shed the remnants of the SARS-CoV-2 RNA virus within the concentrations ∼10-10 copies/L. However, it can decrease up to 10 copies/L in wastewaters due to dilution. Environmental virology and microbiology laboratories play a significant role in the identification and analysis of SARS-CoV-2 ribonucleic acid (RNA) in waste and ambient waters worldwide. Virus extraction or recovery from the wastewater (However, due to lack of knowledge, established procedures, and integrated quality assurance/quality control (QA/QC) approaches, the novel coronavirus RNA investigation for estimating current illnesses and predicting future outbreaks is insufficient and/or conducted inadequately. The present manuscript is a technical review of the various methods and factors considered during the identification of SARS-CoV-2 genetic material in wastewaters and/or sludge, including tips and tricks to be taken care of during sampling, virus concentration, normalization, PCR inhibition, and trend line smoothening when compared with clinically active/positive cases.