Presentation Title
Identification of Borrelia burgdorferi in Field-Collected Ixodes scapularis Ticks from SE Minnesota and West Central Wisconsin using Real-Time qPCR
Abstract
Borrelia burgdorferi, the bacteria that causes Lyme disease, is a spirochete that is present in field-collected Ixodes scapularis ticks. The current assays used to detect the spirochete mainly consist of fluorescent microscopy and traditional PCR, which requires amplification of two separate genes. These procedures are cumbersome, especially when trying to screen large sample sizes. In this study, a protocol was developed using real-time qPCR to detect Borrelia burgdorferi DNA with a much greater sensitivity. The reactions consisted of single reactions with the use of SYBRÒ Green to target and bind DNA with fluorescence. Primers targeting the RecA gene specific to B. burgdorferi was found to be the most effective in the qPCR detection with high success and clear results. Simultaneously, tick DNA was amplified using primers specific to the Ixodes ITS2 gene in order to confirm DNA viability. Further optimization of these protocols are continuing so that 1000’s of ticks collected between 2005-2012 in SE Minnesota and West Central Wisconsin will be tested for presence of Lyme disease.
College
College of Science & Engineering
Department
Biology
Location
Winona, MN
Breakout Room
18
Start Date
4-14-2021 3:00 PM
End Date
4-14-2021 3:45 PM
Presentation Type
Video (Live-Zoom)
Identification of Borrelia burgdorferi in Field-Collected Ixodes scapularis Ticks from SE Minnesota and West Central Wisconsin using Real-Time qPCR
Winona, MN
Borrelia burgdorferi, the bacteria that causes Lyme disease, is a spirochete that is present in field-collected Ixodes scapularis ticks. The current assays used to detect the spirochete mainly consist of fluorescent microscopy and traditional PCR, which requires amplification of two separate genes. These procedures are cumbersome, especially when trying to screen large sample sizes. In this study, a protocol was developed using real-time qPCR to detect Borrelia burgdorferi DNA with a much greater sensitivity. The reactions consisted of single reactions with the use of SYBRÒ Green to target and bind DNA with fluorescence. Primers targeting the RecA gene specific to B. burgdorferi was found to be the most effective in the qPCR detection with high success and clear results. Simultaneously, tick DNA was amplified using primers specific to the Ixodes ITS2 gene in order to confirm DNA viability. Further optimization of these protocols are continuing so that 1000’s of ticks collected between 2005-2012 in SE Minnesota and West Central Wisconsin will be tested for presence of Lyme disease.
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