Faculty, Staff and Student Publications
Language
English
Publication Date
1-1-2025
Journal
Methods in Molecular Biology
DOI
10.1007/978-1-0716-4615-1_27
PMID
40515922
PMCID
PMC13417987
PubMedCentral® Posted Date
7-30-2026
PubMedCentral® Full Text Version
Author MSS
Abstract
The surveillance of SARS-CoV-2 is significantly limited by the presence of numerous variants with distinct behaviors driven by rapid evolution. Although the conventional clinical genome sequencing method provides comprehensive data, it has limitations in capturing the epidemiological dynamics of the variants circulating in the community. In this chapter, we describe a protocol using RBD amplicon sequencing of wastewater samples. We present the detailed experimental procedure for wastewater sample processing, robust nested-PCR-based amplification of the target RBD sequence, and library preparation. Moreover, we describe a general pipeline of sequencing data processing and interpretation, showing the epidemiological and evolutionary trends of SARS-CoV-2 variants in the community.
Keywords
SARS-CoV-2, Wastewater, Humans, COVID-19, Spike Glycoprotein, Coronavirus, Polymerase Chain Reaction, Evolution, Molecular, High-Throughput Nucleotide Sequencing, Genome, Viral
Published Open-Access
yes
Recommended Citation
Xingwen Chen, Chunfu Zheng, and Fuqing Wu, "RBD Amplicon Sequencing and Evolutionary Analysis of SARS-CoV-2 Variants from Wastewater Samples" (2025). Faculty, Staff and Student Publications. 1576.
https://digitalcommons.library.tmc.edu/uthsph_docs/1576