P-45 Wastewater surveillance protocol development and implementation for the recovery of viral fragments of an enveloped virus
Author(s):
K Farmer-Diaz , M Matthew-Bernard , S Cheetham , D Fitzpatrick , K Mitchell , CNL Macpherson , and ME Ramos-Nino
Year of Presentation:
2022
Objective: In the summer of 2021, Grenada experienced its first wave of severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) cases. Wastewater-based epidemiology (WBE) is an effective methodology to monitor the community spread of pathogens and has been implemented on numerous university campuses to identify potential outbreaks. Standardized effective methods to detect enveloped viruses like SARS-CoV-2 are needed to effectively utilize WBE, as most methods implemented have been validated for non-enveloped viruses.
Design and Methods: Pseudomonas syringae bacteriophage (F6) was added to wastewater samples in triplicate to test the efficiency of the VIRuses ADsorption ELution method with two different conditions. Samples were either untreated (NT) or pre-treated with hydrochloric acid (HCl) to a pH of 3.5. The membrane filters were eluted with TrisEDTA-NaCl buffer followed by Trizol RNA extraction and reverse transcription quantitative polymerase chain reaction to quantify viral particles. This filtration method was implemented at a university campus in Grenada
Results: The ratio of F6 Ct .R / PMMoV Ct .R for both treatments was calculated and compared using t-tests to evaluate significant differences. Results showed a mean ratio of 0.75 ± 0.08 for the HCl treated sample compared to 0.86 ± 0.06 for the non-treated sample. The results were statistically significant (p = 0.04).
Conclusion: Membrane filtration using acidification (pH = 3.5) with HCl and elution with Tris-EDTA-NaCl buffer shows to be an effective methodology for the detection of enveloped viruses in WBE. The epidemiological and public health implications of this result will be presented.