Protocol to Longitudinally Quantify SARS-Cov-2 RNA in Wastewater Using RT-Qpcr and Pepper Mild Mottle Virus Normalization
Author Type(s)
Student
Document Type
Article
Publication Date
6-21-2024
DOI
10.1016/j.xpro.2024.103001
Journal Title
STAR Protocols
Keywords
Environmental sciences, Microbiology, Molecular Biology
Disciplines
Medicine and Health Sciences
Abstract
Wastewater surveillance allows severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection levels to be tracked in a community. Here, we present a protocol to longitudinally quantify SARS-CoV-2 RNA in wastewater using quantitative reverse-transcription PCR (RT-qPCR) and pepper mild mottle virus (PMMoV) normalization. We describe steps for the pasteurization of wastewater samples, solids separation, supernatant filtration, viral precipitation and concentration, and RNA extraction. We then detail procedures for RT-qPCR, viral concentration extrapolation, PMMoV normalization, and longitudinal analysis. This protocol has the potential to be used for surveillance of other microorganisms. For complete details on the use and execution of this protocol, please refer to Sanchez Jimenez et al.1
Recommended Citation
Jimenez, B., Sterling, T., Brown, A., Modica, B., Gibson, K., Collins, H., Koch, C., Schwarz, T., & Dye, K. (2024). Protocol to Longitudinally Quantify SARS-Cov-2 RNA in Wastewater Using RT-Qpcr and Pepper Mild Mottle Virus Normalization. STAR Protocols, 5 (2). https://doi.org/10.1016/j.xpro.2024.103001
