Real-time qPCR Assay Development for Detection of Bacillus thuringiensis and Serratia marcescens DNA and the Influence of Complex Microbial Community DNA on Assay Sensitivity
Master of Science (MS)
First Advisor's Name
First Advisor's Committee Title
Second Advisor's Name
Third Advisor's Name
Biology, Forensics, DNA, Real-time qPCR, Biological agents, Detection
Date of Defense
Real-time quantitative polymerase chain reaction (real-time qPCR) assays are an effective technique to detect biological warfare agents and surrogate organisms. In my study, primers were designed to detect chromosomal DNA of biological warfare agent surrogates B. thuringiensis and S. marcescens (representing B. anthracis and Y. pestis, respectively) via real-time qPCR. Species-level specificity of the primers was demonstrated through comparisons with a bacterial strain panel and corroborated by qPCR data. Additionally, the primer efficacy was tested when template DNA was spiked into metagenomic DNA extracted from clinical lung microbiome samples. The results showed that while detection of B. thuringiensis or S. marcescens was still largely successful, the addition of metagenomic DNA did significantly inhibit amplification in most cases. The present study is significant not only for the design of multiple novel primer pairs able to detect bacterial agents in metagenomic DNA, but also the quantitative insight to the influence of background DNA on single species detection at low DNA concentrations.
Segal, Jonathan, "Real-time qPCR Assay Development for Detection of Bacillus thuringiensis and Serratia marcescens DNA and the Influence of Complex Microbial Community DNA on Assay Sensitivity" (2013). FIU Electronic Theses and Dissertations. 1030.
Biology Commons, Microbiology Commons, Molecular Biology Commons
In Copyright. URI: http://rightsstatements.org/vocab/InC/1.0/
This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).