Authors

Ross Bair

Document Type

Honors Project

First Advisor

Dr. Stephen Baron

Degree Award Date

Fall 2001

Keywords

Polymer Chain Reaction, Detection, Lyme Disease Bacterium, Deer Ticks

Disciplines

Biology | Immunology and Infectious Disease | Laboratory and Basic Science Research

Abstract

In 1999 as part of a student project Polymerase chain reaction methods were used to amplify a segment of Borrelia burgdorferi DNA using primers for the ospA and ospB genes. To adequately assess the ability of a PCR detection of the bacterium serial dilutions of DNA were made and subjected to PCR using the ospA and ospB primers. We detected the predicted 925 hp PCR product when using a DNA template concentration of as low as 444 pg/μI. Additionally, the primers used for the PCR were run against genomic DNA extracted from 6 other bacteria including two bacterial isolates extracted from a tick itself. These results showed absence of the 925bp PCR product indicating that the primers are specific. We were unable to successfully culture the Borrelia burgdorferi bacteria and methods of detecting a DNA band after the tick crushing and DNA extraction were unsuccessful even with modified procedures that are documented to work.

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