Discovering new viral lineages and estimating their abundance in wastewater
MetadataShow full item record
Wastewater surveillance of SARS-CoV-2 has emerged as a critical tool for tracking the spread of COVID-19. In addition to estimating the relative case numbers using qPCR, SARS-CoV-2 genomic RNA can be extracted from wastewater and sequenced. The sequenced genomes provide information about which lineages, in particular which variants of concern (VOCs) are present in a community. Wastewater RNA sequencing data has two distinct challenges: First, the genomes are highly fragmented and the alignments often have poor genome coverage. Second, the samples are comprised of a mixture of genomes so mutations cannot be directly attributed to a single lineage. In this thesis, I explore methods to overcome these two challenges to extract useful information from the samples. First, I look at the problem of determining the relative abundance of VOCs. Most existing techniques only consider mutations which are unique to a particular VOC which massively reduces the amount of usable data. I introduce a new technique which extends mean and median frequencies over shared mutations in order to make use of the huge pool of shared mutations. Next, I investigate strategies for designing single-amplicon sequencing methods. I look at selecting single amplicons which are well-conserved and rich in information. I also design a single amplicon which is capable of amplifying multiple coronaviruses. I conclude the SARS-CoV-2 work by providing a technique which can identify novel lineages and sublineages from wastewater sequencing runs. Finally, I show that the techniques for analyzing SARS-CoV-2 in wastewater can also be applied to an important plant pathogen, the Tomato Brown Rugose Fruit Virus.
Cite this version of the work
Isaac Ellmen (2022). Discovering new viral lineages and estimating their abundance in wastewater. UWSpace. http://hdl.handle.net/10012/18818