DEVELOPMENT OF A NOVEL QUANTITATIVE PCR PROTOCOL FOR IMPROVED BORRELIA BURGDORFERI DETECTION IN MAMMAL TISSUE
Author: Nicholas Fried
Major: Animal and Veterinary Sciences
Graduation Year: 2016
Thesis Advisor: James A. Weber
Description of Publication: Developing an understanding of Borrelia burgdorferi infection patterns in mammal reservoirs is imperative for combatting Lyme disease. The most important of these reservoirs is the white-footed mouse. This study aimed to discern the B. burgdorferi infection prevalence in 83 mammal tissue biopsies collected in the summer of 2010 from Cape Cod, Massachusetts. Infection prevalence was measured through a qPCR assay and compared to results from a similar study in Fort McCoy, Wisconsin. Presence of an unknown PCR inhibitor in mammal tissue samples required development and validation of a novel qPCR protocol to improve detection of the pathogen. Bovine serum albumin at various concentrations, through extensive testing of Borrelia spiked samples, met this requirement. Mammal tissue biopsy qPCR with 100 ng/μL of BSA was shown to be equally as effective as xenodiagnostic testing for the detection of B. burgdorferi. Overall mammalian infection with B. burgdorferi in Cape Cod was approximately 50 percent, with white-footed mouse infection prevalence of 60 percent. Overall infection in Cape Cod differed statistically (p = 0.06) from the observed value in Fort McCoy. There was no significant difference between the white-footed mouse infection prevalence in the two regions.
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