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.
","abstract_html":"<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>","abstract_has_math":false,"creators":["Peacock, Tess"],"institution":null,"degree_name":"Master of Science","degree_level":"Thesis","degree_discipline":"Biology","degree_department":null,"school":null,"contributors":["Josh Sharp, Ph.D"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-08-01T07:00:00Z","date_published":"2019-08-01T07:00:00Z","updated_at":"2026-07-24T03:24:12Z","subjects":["bacteria","rapid identification","diagnostics","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://commons.nmu.edu/theses/599","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Josh Sharp, Ph.D"]},{"key":"dc:creator","label":"Author","values":["Peacock, Tess"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2024-07-09T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["bacteria","rapid identification","diagnostics","Medicine and Health Sciences"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://commons.nmu.edu/theses/599"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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.
"]},{"key":"dc:title","label":"Title","values":["DEVELOPMENT OF A RAPID DIAGNOSTIC ASSAY FOR THE COLORIMETRIC DETECTION OF LEGIONELLA BACTERIA USING LOOP MEDIATED ISOTHERMAL AMPLIFICATION"]}]}],"canonical_facts":{"dc:contributor":["Josh Sharp, Ph.D"],"dc:creator":["Peacock, Tess"],"dc:date.available":["2024-07-09T07:00:00Z"],"dc:description.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.
"],"dc:identifier":["https://commons.nmu.edu/theses/599"],"dc:subject":["bacteria","rapid identification","diagnostics","Medicine and Health Sciences"],"dc:title":["DEVELOPMENT OF A RAPID DIAGNOSTIC ASSAY FOR THE COLORIMETRIC DETECTION OF LEGIONELLA BACTERIA USING LOOP MEDIATED ISOTHERMAL AMPLIFICATION"],"thesis:degree_discipline":["Biology"],"thesis:degree_level":["Thesis"],"thesis:degree_name":["Master of Science"]},"updated_at":"2026-07-24T03:24:12Z"}