Development of qPCR Based Method to Quantify Colonization of an Engineered Bacterium in Gut

Thumbnail Image
Supplemental Files
McMullen, Alexandra (Alex)
Major Professor
Anumantha Kanthasamy
Committee Member
Journal Title
Journal ISSN
Volume Title
Research Projects
Organizational Units
Organizational Unit
Biomedical Sciences

The Department of Biomedical Sciences aims to provide knowledge of anatomy and physiology in order to understand the mechanisms and treatment of animal diseases. Additionally, it seeks to teach the understanding of drug-action for rational drug-therapy, as well as toxicology, pharmacodynamics, and clinical drug administration.

The Department of Biomedical Sciences was formed in 1999 as a merger of the Department of Veterinary Anatomy and the Department of Veterinary Physiology and Pharmacology.

Dates of Existence

Related Units

  • College of Veterinary Medicine (parent college)
  • Department of Veterinary Anatomy (predecessor, 1997)
  • Department of Veterinary Physiology and Pharmacology (predecessor, 1997)

Journal Issue
Is Version Of
Biomedical Sciences

Parkinson’s Disease is the leading neurodegenerative disease in the western world, caused by degeneration of neurons in the substantia nigra. This causes a decrease of dopamine in nigrostriatal neural circuitry, leading to motor and nonmotor symptoms which can be incredibly debilitating for the patient. Current dopamine supplementation therapy does not provide a steady state neurotransmitter level and can lead to dyskinesias and dystonia’s. However, Escherichia coli Nissle (EcN) bacteria have been shown to prove effective therapeutic treatments that can be engineered to specifically treat certain diseases, such as Phenylketonuria. EcN can be engineered to upregulate production of L-DOPA by adding an extra HpaBC gene. Administration of L-DOPA through ECN can maintain steady state dopamine level and thereby normalizing dopaminergic neural pathways for patients suffering from Parkinson’s Disease. The goal of this study was to monitor EcN-DOPA gut colonization in mouse models of Parkinson’s disease using a qPCR method. We generated a unique primer set for ECN-L-DOPA and determined its utility in measuring the colonization. Our results showed that ECN-L-DOPA primer effectively amplifies the gene and detected the engineered bacteria in the fecal sample. Further evaluation in Mitopark transgenic mouse model of Parkinson’s disease, qPCR method successfully detected colonization profile of ECN-DOPA and correlated with the plasma level with plasma L-DOPA level. Collectively, our results show qPCR method is very amenable for monitoring gut colonization in microbiome-based therapy.

Tue Jan 01 00:00:00 UTC 2019