Using qPCR to detect trace amounts of pathogens
Start Date
August 2025
End Date
August 2025
Location
ALT 302
Abstract
Escherichia coli is a type of bacteria which lives within the intestines of warm-blooded animals and is often used as a biological marker to show fecal contamination within water. The presence of E. coli in water is dangerous as numerous health issues can arise from drinking or swimming in contaminated water. Traditional methods used for detecting E. coli include culturing on differential and selective media. However, culturing is time-consuming and has low sample throughput. Alternatively, traditional PCR can be used to quickly identify bacterial species by detecting specific subsets of genes. After the use of both prior techniques, quantitative PCR (qPCR) which has the same characteristics as regular PCR, while also providing a relative number of bacteria present in the sample. In this project, regular and qPCR has been used to verify primers and test for the CydA, lacY, and yDiv genes in control lab samples and Shiga Toxin producing E. coli. (STEC) From this point on, qPCR will be used on environmental samples to detect and determine if STEC is in local waterways.
Using qPCR to detect trace amounts of pathogens
ALT 302
Escherichia coli is a type of bacteria which lives within the intestines of warm-blooded animals and is often used as a biological marker to show fecal contamination within water. The presence of E. coli in water is dangerous as numerous health issues can arise from drinking or swimming in contaminated water. Traditional methods used for detecting E. coli include culturing on differential and selective media. However, culturing is time-consuming and has low sample throughput. Alternatively, traditional PCR can be used to quickly identify bacterial species by detecting specific subsets of genes. After the use of both prior techniques, quantitative PCR (qPCR) which has the same characteristics as regular PCR, while also providing a relative number of bacteria present in the sample. In this project, regular and qPCR has been used to verify primers and test for the CydA, lacY, and yDiv genes in control lab samples and Shiga Toxin producing E. coli. (STEC) From this point on, qPCR will be used on environmental samples to detect and determine if STEC is in local waterways.
