Detection of SARs-CoV-2 in wastewater, using the existing environmental surveillance network: An epidemiological gateway to an early warning for COVID-19 in communities

Abstract

The ongoing COVID-19 pandemic caused by SARs-CoV-2 was considered to be transmitted person to person via droplet infections and fecal-oral transmission. To determine this, we used the existing polio environment surveillance network in Pakistan to investigate presence of SARs-CoV-2 using three commercially available kits and E-Gene detection published assay for surety and confirmatory of positivity. A Two-phase separation method is used for sample clarification and concentration. Before proceeding directly for RNA extraction, an additional high-speed centrifugation (14000Xg for 30 min) step was introduced to increase viral RNA yield resulting decrease in Cq value. A total of 78 wastewater samples collected from 38 districts across Pakistan, 74 wastewater samples from existing polio environment surveillance sites, 3 from drains of COVID-19 infected areas and 1 from COVID 19 quarantine center drainage, were tested for presence of SARs-CoV-2. 21 wastewater samples (27%) from 13 districts turned to be positive on RT-qPCR. SARs-COV-2 RNA positive samples from areas with COVID patients and COVID 19 patient quarantine center drainage strengthen the findings and use of wastewater surveillance in future. Furthermore, sequence data of partial ORF 1a generated from COVID 19 patient quarantine center drainage sample also reinforce our findings that SARs-CoV-2 can be detected in wastewater. This study finding indicates that SARs-CoV-2 detection through wastewater surveillance has an epidemiologic potential that can be used as early warning system to monitor viral tracking and circulation in cities with lower COVID-19 disease burden or heavily populated areas where door-to-door tracing may not be possible. However, attention needed on virus concentration and detection assay to increase the sensitivity. Development of highly sensitive assay will be an indicator for virus monitoring and to provide early warning signs.

Positive environmental effects of the coronavirus 2020 episode: a review

“…Hart and Halden (2020) conducted a computational modeling study and cost analysis survey to confirm the effectiveness of WBE in tracking COVID-19; they suggested that this tool requires to consider the temperature effects as well. Sharif et al (2020) also applied the existing polio Fig. 8 Venice lagoons and waterways quality (top) in 2020 and (bottom) after a month pandemic lockdown in 2019.…”

A review on presence, survival, disinfection/removal methods of coronavirus in wastewater and progress of wastewater-based epidemiology

“…Although there is a lack of studies on the presence of SARS-CoV-2 in wastewater, the detection of viral RNA (ribonucleic acid) in wastewater samples has gained considerable attention based on the idea of wastewater-based surveillance to detect community spread. As per the aim and scope of the present study, 34 works (eight research articles ( Table 1 ) and 26 pre-print articles [ [18] , [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] , [31] , [32] , [33] , [34] , [35] , [36] , [37] , [38] , [39] , [40] , [41] , [42] , [43] ]) are included to understand the applicability of wastewater-based surveillance.…”

“…Döhla et al [ 42 ] observed 10 out of 66 wastewater samples as positive collected from toilets, showers, and washbasin of quarantined households. Sharif et al [ 43 ] collected 78 wastewater samples (74 from polio surveillance sites, 3 from drains of affected area, 1 from drainage of quarantine center) out of which 21 samples were tested positive for the presence of SARS-CoV-2 RNA.…”

COVID-19 Crisis Creates Opportunity towards Global Monitoring & Surveillance

“…Long-term, inexpensive surveillance of SARS-CoV-2 would help public health agencies to implement appropriate measures and governments to shape their economies, and the key to that is employing cost-effective and non-invasive surveillance strategies [161]. Environmental microbiologists have investigated pathogens such as waterborne, foodborne and faecal-oral viruses or enteric viruses such as norovirus, hepatitis A virus, and poliovirus from the sewage excreted through faeces and used it as a public health surveillance tool [137,[162][163][164][165][166][167][168][169], and more recently for monitoring SARS-CoV-2 [115,140,142,144,[170][171][172][173][174][175].…”

“…They suggested that quantitative monitoring of SARS-CoV-2 genomes in wastewater should give further and pivotal information for better surveillance of SARS-CoV-2 circulation at the local or regional scale. In Pakistan, Sharif et al [175] found that 21 wastewater samples (27%) from 13 districts were PCR positive, indicating that wastewater surveillance has an epidemiologic potential which could be considered to be an early warning system for monitoring viral tracking in different districts.…”