Quantification of Gordonia terrae Bacteriophages using Lysate qPCR

Date of Award

5-8-2024

Document Type

Thesis

Department

Biology

First Reader

Dr. Ruth Plymale

Second Reader

Dr. Christin Pruett

Third Reader

Professor Hallie Wallace

Abstract

Bacteriophages are found abundantly throughout the earth and are viruses that are able to infect and kill bacterial cells. They are being used in various ways in the medical field and in research labs across the world. The current method, the double-layer agar plating method, is commonly practiced to detect and quantify bacteriophages, and it is a strenuous two-to-three-day process if completed without errors. A quicker method to detect bacteriophages is needed. Quantitative Polymerase Chain Reaction can amplify DNA in approximately three hours. The double-layer agar plating method was compared to qPCR using one-fifth dilutions of bacteriophage GrandSlam lysate to determine which method quantified the bacteriophage most accurately and efficiently. The results of the lysate qPCR revealed this highly sensitive test counts all bacteriophages present, whether they are fully intact and able to reproduce or not. The double-layer agar plating method only produces plaques from bacteriophages that have the ability to infect bacterial cells. The bacteriophages must have their head and tail connected to infect cells. The lysate qPCR does not seem to be able to replace the double-layer agar plating method due to not having the ability to only quantify bacteriophage DNA copies that are able to infect and reproduce in bacterial cells.

Comments

This thesis is currently embargoed. It will be available May 2029.

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