Discovery of DNA Aptamers as Probes for Detecting Ranavirus MCP
Presentation Type
Poster
Faculty Advisor
Nina Goodey
Access Type
Event
Start Date
26-4-2023 11:00 AM
End Date
26-4-2023 12:00 PM
Description
Global amphibian populations have been declining since the 20th century; one of the most significant contributors to these declines is a genus of viral pathogens known as Ranavirus (Rv). Frog virus 3 is one of these viral agents and is currently found affecting frog populations throughout NJ, MD, PA, DE, and VA. In the absence of a cure, it is imperative to be able to detect and monitor the virus in the wild, but current testing methods require extensive training, expensive equipment, and are time consuming. To deal with these issues and develop a rapid field test that provides real time results, we have used a DNA aptamer discovery technique to generate short single-stranded DNA sequences that specifically bind to the viral major capsid protein (MCP). This technique, known as SELEX (Systematic Evolution of Ligands by Exponential Enrichment), is a polymerase chain reaction (PCR) based process that narrows a large library of random DNA sequences into a smaller library of targeted sequences. By performing SELEX and sequencing the resulting aptamers, we have enriched aptamers for binding to Rv MCP. Such aptamers will be screened using electrophoretic mobility shift assays to assess their binding affinity to MCP, to find aptamers for use in the proposed rapid field tests, as well as identify sequence patterns necessary to bind to the viral protein.
Discovery of DNA Aptamers as Probes for Detecting Ranavirus MCP
Global amphibian populations have been declining since the 20th century; one of the most significant contributors to these declines is a genus of viral pathogens known as Ranavirus (Rv). Frog virus 3 is one of these viral agents and is currently found affecting frog populations throughout NJ, MD, PA, DE, and VA. In the absence of a cure, it is imperative to be able to detect and monitor the virus in the wild, but current testing methods require extensive training, expensive equipment, and are time consuming. To deal with these issues and develop a rapid field test that provides real time results, we have used a DNA aptamer discovery technique to generate short single-stranded DNA sequences that specifically bind to the viral major capsid protein (MCP). This technique, known as SELEX (Systematic Evolution of Ligands by Exponential Enrichment), is a polymerase chain reaction (PCR) based process that narrows a large library of random DNA sequences into a smaller library of targeted sequences. By performing SELEX and sequencing the resulting aptamers, we have enriched aptamers for binding to Rv MCP. Such aptamers will be screened using electrophoretic mobility shift assays to assess their binding affinity to MCP, to find aptamers for use in the proposed rapid field tests, as well as identify sequence patterns necessary to bind to the viral protein.