Date of Award

8-2019

Degree Type

Thesis

Degree Name

Master of Science

Department

Biology

Program

Biology

First Advisor/Chairperson

Josh Sharp, Ph.D

Abstract

Legionella are aquatic bacteria capable of inhabiting man-made water systems. Inhaling water droplets carrying Legionella can cause Legionnaires’ disease, a rapidly progressing pneumonia. From 2000-2016 the number of Legionnaires’ cases increased 5-fold. Detecting Legionella in water systems can be challenging. Legionella have complex growth requirements, requiring expensive specialized media to grow. It can take 10 days to grow Legionella colonies on agar plates. There are few rapid assays available that can identify multiple Legionella species. This project aims to develop a Loop Mediated Isothermal Amplification (LAMP) assay that can detect multiple species of Legionella in less than one hour. LAMP utilizes four primers, a strand displacing DNA polymerase, and a single incubation temperature to amplify DNA from a target specimen. Four primers were designed that amplify a unique Legionella 16S rDNA sequence.

We have demonstrated that a Legionella specific LAMP assay amplified only DNA from Legionella. When DNA from Legionella was amplified the pH in the reaction decreased. The pH drop triggered a color change of a pH indicator present in the reaction. Some LAMP assays have been shown to detect as few as 10-12 copies of DNA in food or water samples. Colorimetric Legionella LAMP can detect as few as two copies of DNA in under 90 minutes. The colorimetric LAMP assay can be used as a fast and accurate method for detecting Legionella in water samples and for monitoring purposes, as well as for tracing sources of outbreaks.

Access Type

Open Access

Justification for Restricting Access

The worked contained within the thesis will be submitted for publication in a peer reviewed journal by 7/31/21. The embargo is required to protect the data so that no one else can copy Tess Peacock's original work and publish it. Additionally, some journals require that the data submitted can not published elsewhere prior to submission to their journal. Publication in the NMU Commons could violate that condition.

Available for download on Tuesday, July 09, 2024

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