Accurately Quantifying Stage Specific Enolase and GAPDH Transcripts in Toxoplasma gondii
Advisor Information
Paul Davis
Location
Milo Bail Student Center Ballroom
Presentation Type
Poster
Start Date
8-3-2013 9:00 AM
End Date
8-3-2013 12:00 PM
Abstract
Toxoplasma gondii is an intracellular parasite with the ability to infect humans and animals, and is closely related to the parasite that causes malaria. The tachyzoite form of the parasite causes infection throughout the body, but after detection by the immune system, T. gondii can form long-lived, drugresistant cysts called bradyzoites in brain and muscle tissue. Present laboratory protocols are not able to produce significant bradyzoite cultures, compromising the ability to evaluate potential drug compounds effective against the bradyzoite stage. We hypothesized that the effectiveness of the growth medium in transforming Toxoplasma gondii parasites from the tachyzoite to the bradyzoite stage can be rapidly and precisely determined by using quantitative PCR. Development of unique forward and reverse primers, along with a fluorescent probe for enhanced target analysis, allowed us to detect relative differences in parasite life stage, based on measuring transcription rates of tachyzoite-specific and bradyzoite-specific gene expression. These methods successfully quantified bradyzoite amounts, which can be used in further experiments to aid in drug selection against the bradyzoite stage.
Accurately Quantifying Stage Specific Enolase and GAPDH Transcripts in Toxoplasma gondii
Milo Bail Student Center Ballroom
Toxoplasma gondii is an intracellular parasite with the ability to infect humans and animals, and is closely related to the parasite that causes malaria. The tachyzoite form of the parasite causes infection throughout the body, but after detection by the immune system, T. gondii can form long-lived, drugresistant cysts called bradyzoites in brain and muscle tissue. Present laboratory protocols are not able to produce significant bradyzoite cultures, compromising the ability to evaluate potential drug compounds effective against the bradyzoite stage. We hypothesized that the effectiveness of the growth medium in transforming Toxoplasma gondii parasites from the tachyzoite to the bradyzoite stage can be rapidly and precisely determined by using quantitative PCR. Development of unique forward and reverse primers, along with a fluorescent probe for enhanced target analysis, allowed us to detect relative differences in parasite life stage, based on measuring transcription rates of tachyzoite-specific and bradyzoite-specific gene expression. These methods successfully quantified bradyzoite amounts, which can be used in further experiments to aid in drug selection against the bradyzoite stage.