Office of Undergraduate Research

Permanent URI for this communityhttp://hdl.handle.net/1903/20157

Emphasizing equitable and inclusive access to research opportunities, the University of Maryland's Office of Undergraduate Research (OUR) empowers undergraduates and faculty to engage and succeed in inquiry, creative activity, and scholarship. This collection includes materials shared by undergraduate researchers during OUR events. It also encompasses materials from Undergraduate Research Day 2020, Undergraduate Research Day 2021, and Undergraduate Research Day 2022, which were organized by the Maryland Center for Undergraduate Research.

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Author: search.filters.author.Kaiser, Jillian

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    One-Pot Ligation LAMP Assay to Detect miRNA-222: A Glioma Biomarker
    (2024) Pallavajjala, Roshni; Adane, Yedidya; Fernandes, Catarina; Kaiser, Jillian; Patel, Khushi; Spirito, Catherine
    Many cancer diagnostic methods are invasive, time-consuming, and expensive. Delayed cancer diagnosis can lower patient survival rates. PCR-based techniques that detect miRNA biomarkers in blood have been utilized as early screening tools for various cancers. As an alternative to PCR, we designed and optimized an isothermal amplification technique, Ligation Loop-Mediated Isothermal Amplification (Ligation LAMP) assay, to detect miR-222, an established biomarker that is found in elevated levels in the bloodstream of early-stage glioma patients. We designed colorimetric and fluorescent Ligation-LAMP assays and demonstrated their specificity and sensitivity in detecting miR-222. We are working on implementing our assay into a One-Pot system, using Thermally Responsive Alkane Partitions (TRAPs) and a strand displacement assay utilizing magnetic beads. We found that the Ligation LAMP assay is sensitive and specific to glioma biomarker miR-222 and different probe lengths for Strand Displacement did not have a significant impact on ligation. With these results, we can further improve the One-Pot assay to be more point-of-care.
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    Ligation-LAMP to Detect miR-221 Cancer Biomarker
    (2024) Kaiser, Jillian; Lee, HaeSung Lola; Patel, Khushi; Sugg, Bethany; Spirito, Catherine
    Breast cancer is the second most common cancer in women and can be fatal. Current breast cancer diagnostic tools such as mammograms, MRIs, and biopsies are invasive, costly, time-consuming, and inefficient. Isothermal amplification assays, such as loop-mediated isothermal amplification (LAMP), can be designed to detect miRNA cancer biomarkers in blood samples. The focus of our project is to detect miRNA-221, a breast cancer biomarker, using a ligation-LAMP assay with high specificity and sensitivity. During ligation LAMP, miR-221 is reverse transcribed to cDNA. The cDNA acts as a splint to ligate two hairpins that form the dumbbell needed for LAMP amplification by Bst Polymerase. LAMP amplification can be detected through colorimetric methods, using phenol red and HNB. Intercalating fluorescent dye can also be used, or a CRISPR system can be used and yield a fluorescent output as well. Our primary findings include successful tests using colorimetric LAMP. We have seen nonspecific amplification causing false positives, but by adding betaine, a reagent that hinders nucleic acid amplification rate, to our reactions, the positive control amplifies sooner than the negative. Using the colorimetric and fluorescent methods described, ligation LAMP can be used to efficiently detect breast cancer biomarker miR-221.