Microbial Source Tracking Method Validation to Identify Hosts of Fecal Contamination and Pathogen Presence in Source and Stored Drinking Water in Urban Kenya

middle- income countries. These pathogens can spread from feces to hosts through multiple

different pathways making sanitation interventions ineffective at completely limiting the spread

of disease. Molecular source tracking (MST) assays have previously been used for the

identification of host-specific feces in water and other environmental samples. For this project,

human and animal fecal samples along with stored and source water samples were collected from

individual homes in an urban area of Kenya. Prior to the use of the MST assays, the nucleic acid

extraction methods were validated to assure the quality of the RNA and DNA extracted from the

samples and their suitability for downstream applications such as qPCR. Additionally, the

extraction methods were validated for use in the Taqman Array Card system which uses

microfluidic assays for the detection of specific enteric pathogens. These results will aid in the

overall evaluation of the sources of fecal contamination and pathogens present in the collected

water samples. Additionally, a systematic literature review was conducted to examine the extent

to which MST assays have been used to detect fecal contamination in low- and middle-income

countries.