6th Annual Judith Ramaley Celebration of Research and Creative Scholarship
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Poster #112 Development of Real-Time PCR Protocol to Simultaneously Detect Lyme Disease and Babesiosis Erica Tipcke, Alice Kamau, Ashley Piper, and Michael Lehrke Faculty Mentor: Kimberly Bates Lyme disease and Babesiosis are the two tick-borne illnesses being studied in this research because they are considered to be emerging infectious diseases. The main vector in North America for both these organisms is the Ixodes scapularis tick. Isolated DNA of infected I. scapularis may contain Borrelia burgdorferi and/or Babesia microti sequences. The objective of this research is to focus on the prevalence of both diseases in southeast Minnesota and west central Wisconsin. Ticks were harvested from white-tailed deer (Odocoileus virginianus) from both these regions of Minnesota and Wisconsin. After sorting and extracting the DNA, polymerase chain reaction (PCR) was used to detect any B. burgdorferi and B. microti. A multiplex real-time PCR (qPCR) assay is being developed to enhance the testing time of the I. scapularis DNA. The advantage to multiplex qPCR is that two or more genes can be detected at once, which allows for the detection of B. burgdorferi and B. microti within the same test run. Different primers and probes were tested on standard PCR to ensure detection of the correct genes before implementing qPCR testing of I. scapularis DNA. The specificity and sensitivity of qPCR will be compared to the standard PCR results. Using qPCR to detect the genes in the I. scapularis DNA allows for the confirmation of the prevalence of I. scapularis ticks carrying one or both of the diseases. This information can be important for use by scientists, doctors, the general public, and within the education system. |
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