Presenter(s)
Anthony Appicelli and Jenna Junker
Abstract
Black-legged ticks (Ixodes scapularis) and Lyme disease, caused by the bacterium Borrelia burgdorferi, are prevalent in the upper Midwest. Accurate determination of the percentage of Ixodes ticks that carry Lyme disease could help physicians and health experts develop treatment and prevention plans to curb the prevalence of disease. The purpose of this study was to see if accurate results for both Ixodes DNA and Borrelia DNA could be analyzed in the same qPCR assay for faster Lyme disease testing. Current testing in humans is achieved by running two separate diagnostic assays, an ELISA and Western Blot, both of which are very time-consuming and temperamental lab procedures. By using qPCR, an automated result is achieved in a shorter amount of time with less errors because of the sensitivity of the assay. Throughout the two-semester long experiment, several adjustments were made to increase the accuracy of the qPCR, including changes to the primers, their concentrations, and DNA content. Additionally, the thermocycler conditions were frequently modified to accommodate the simultaneous use of both the Ixodes specific and Borrelia-specific primers for running the test. After nine months of trial, the results were inconclusive. This study encountered many difficulties, like contaminations in the primers that produced false positive results and condition complications causing both false positive and negative results. Currently, additional modifications are being assessed to improve the results of the qPCR.
College
College of Science & Engineering
Department
Biology
Campus
Winona
First Advisor/Mentor
Kimberly Bates
Start Date
4-24-2025 1:00 PM
End Date
4-24-2025 2:00 PM
Presentation Type
Poster Session
Format of Presentation or Performance
In-Person
Session
2a=1pm-2pm
Poster Number
23
Included in
Development of a qPCR for detecting Borrelia burgdorferi and Ixodes tick DNA simultaneously
Black-legged ticks (Ixodes scapularis) and Lyme disease, caused by the bacterium Borrelia burgdorferi, are prevalent in the upper Midwest. Accurate determination of the percentage of Ixodes ticks that carry Lyme disease could help physicians and health experts develop treatment and prevention plans to curb the prevalence of disease. The purpose of this study was to see if accurate results for both Ixodes DNA and Borrelia DNA could be analyzed in the same qPCR assay for faster Lyme disease testing. Current testing in humans is achieved by running two separate diagnostic assays, an ELISA and Western Blot, both of which are very time-consuming and temperamental lab procedures. By using qPCR, an automated result is achieved in a shorter amount of time with less errors because of the sensitivity of the assay. Throughout the two-semester long experiment, several adjustments were made to increase the accuracy of the qPCR, including changes to the primers, their concentrations, and DNA content. Additionally, the thermocycler conditions were frequently modified to accommodate the simultaneous use of both the Ixodes specific and Borrelia-specific primers for running the test. After nine months of trial, the results were inconclusive. This study encountered many difficulties, like contaminations in the primers that produced false positive results and condition complications causing both false positive and negative results. Currently, additional modifications are being assessed to improve the results of the qPCR.
