DEVELOPMENT OF A RAPID DIAGNOSTIC ASSAY FOR THE COLORIMETRIC DETECTION OF LEGIONELLA BACTERIA USING LOOP MEDIATED ISOTHERMAL AMPLIFICATION

<p><em>Legionella </em>are aquatic bacteria capable of inhabiting man-made water systems. Inhaling water droplets carrying <em>Legionella </em>can cause Legionnaires’ disease, a rapidly progressing pneumonia. From 2000-2016 the number of Legionnaires’ cases increased 5-fold. Detecting <em>Legionella </em>in water systems can be challenging. <em>Legionella </em>have complex growth requirements, requiring expensive specialized media to grow. It can take 10 days to grow <em>Legionella </em>colonies on agar plates. There are few rapid assays available that can identify multiple <em>Legionella </em>species. This project aims to develop a Loop Mediated Isothermal Amplification (LAMP) assay that can detect multiple species of <em>Legionella </em>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 <em>Legionella </em>16S rDNA sequence.</p> <p>We have demonstrated that a <em>Legionella </em>specific LAMP assay amplified only DNA from <em>Legionella</em>. When DNA from <em>Legionella </em>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 <em>Legionella </em>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 <em>Legionella </em>in water samples and for monitoring purposes, as well as for tracing sources of outbreaks.</p>

Name thesis:degree_name

Master of Science

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Biology

Year dc:date.available

2019

Author dc:creator

• Peacock, Tess

Contributors dc:contributor

• Josh Sharp, Ph.D

• bacteria
• rapid identification
• diagnostics
• Medicine and Health Sciences

Repository record dc:identifier

https://commons.nmu.edu/theses/599

OAI identifier oai:identifier

oai:commons.nmu.edu:theses-1629