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NOVEL MICROFLUIDIC DEVICE FOR IN-FIELD PURIFICATION, AMPLIFICATION, AND DETECTION OF DNA FROM MYCOTOXIN-PRODUCING FUNGI

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thesis
posted on 2023-03-08, 15:22 authored by Cole Benjamin ReynoldsCole Benjamin Reynolds

Mycotoxins are toxic molecules produced by fungi and are harmful to human health when ingested. Mycotoxins produced by fungi that infect food crops can enter the food stream directly in manufactured produces (foods like bread) or indirectly through animal feed. Detection of mycotoxins is commonly performed after harvesting and processing of the crops because mycotoxins can increase in concentration during these steps. However, if farmers could detect the toxin-forming fungi prior to harvesting, growers can reduce cross-contamination, save wasted resources used in harvesting and processing of infected crops, and increase food safety and security. Fusarium graminearum is a fungal species that infects cereal crops and produces deoxynivalenol (DON), a harmful mycotoxin called vomitoxin. Because Fusarium graminearum has a large economic impact in the United States and globally on wheat and barley markets, it was selected as a proof-of-concept for the detection assay. For detection of fungus to be useful in determining infected areas of fields, the device must be inexpensive and portable. A low-cost, portable microfluidic device was designed to both extract the fungal DNA from wheat samples and perform a fluorescence-based loop-mediated isothermal amplification (LAMP) assay to detect the fungal DNA. Contaminated wheat samples were used to demonstrate the effectiveness of the device. The device was able to detect Fusarium graminearum in the wheat samples at concentrations relevant to DON concentration limits set forth by the U.S. Food and Drug Administration (FDA). Detection of multiple mycotoxin-producing fungal species is possible through the addition of more LAMP assays for multiplexing specific target genes.

History

Degree Type

  • Master of Science in Biomedical Engineering

Department

  • Biomedical Engineering

Campus location

  • West Lafayette

Advisor/Supervisor/Committee Chair

J Paul Robinson

Additional Committee Member 2

Euiwon Bae

Additional Committee Member 3

Jacqueline Linnes

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