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)

Comments

Poster file was not uploaded for this entry.

This document is currently not available here.

Share

COinS
 
Apr 14th, 3:00 PM Apr 14th, 3:45 PM

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.