Assessment of Qualitative PCR Protocol on Ixodes scapularis for the Identification of Borrelia burgdorferi

Ixodes scapularis, more commonly known as a black-legged tick or a deer tick, is the vector for Lym disease (Borrelia burgdoreferi) in much of the United States. Their definitive host is deer (Family Cervidae), which means they are found in climates where deer thrive. These areas include the eastern and norther Midwest of the United States along with southeastern Canada. The ticks are known for transmitting Lyme disease to humans (Homo sapiens), canines (Canis lupus familiaris), and other mammals. Lyme disease is caused by Borrelia burgdorferi which is a spirochete bacterium. The purpose of this study was to determine the prevalence of infected ticks in the Southeast MN and West Central Wisconsin areas on either side of the Mississippi river using Real-Time or Quantitative PCR (qPCR). DNA was extracted from approximately 6,000 ticks that were collected between 2005 and 2012 from legally harvested white-tailed deer (Odocoileus virginianus) from Buffalo County, WI and Winona County, MN. The DNA was analyzed using quantitative PCR. An iTaq Universal SYBR green supermix was used along with a RecAF and RecAR primers for the Borrelia DNA and ITS2F and ITS2R were used to amplify tick DNA. The amplifications that occurred from ITS2F and ITS2R were used to ensure that the tick DNA was viable. For RecAR and RecAF, the amplifications determined whether the tick had B. burgdorferi or not. Results to date show that the unknown tick DNA is not amplifying consistently when being tested for the presence of B. burgdoreferi. The next step in the experiment is to determine how to amplify the DNA more consistently and to continue testing ticks for the presence of B. burgdoreferi collected between 2005 and 2012.