The COVID-19 outbreak highlighted the important role that diagnostic tests play in the healthcare system. To reduce the impact of infectious disease outbreaks, the development of rapid and cost-effective point-of-care-tests (POCTs) is crucial. With the dissemination of the Monkeypox (Mpox) virus, it became a necessity to produce POCTS that are inexpensive and easy to use. This work explored the construction of two colorimetric assays that aim to detect Mpox genetic signatures. One is based on the split-peroxidase-like deoxyribozyme probes (sPDz), while the other utilizes a cascade system of split RNA-cleaving deoxyribozyme (sDz) and peroxidase-like deoxyribozyme (PDz). Both rely on catalytic probes as well as a G-Quadruplex (G4) structure to facilitate the production of a color change in the presence of the genetic signatures of Mpox. The sPDz probes were initially tested with synthetic genomic fragments of Mpox and other Orthopoxviruses for selectivity purposes. The sPDz probes were then further optimized. The optimal sPDz probes and the sDz/PDz cascade system were tested with an amplified genome fragment of Mpox. The genome fragments were generated by using both symmetric and asymmetric polymerase chain reaction (PCR). With further optimization to increase the signal-to-background ratio, these probes may show promise as an assay that may have the potential to be incorporated to develop POCTs.
Bachelor of Science (B.S.)
College of Sciences
Ahn, Jaehyun, "Split Catalytic Probes for the Detection of Monkeypox Virus" (2023). Honors Undergraduate Theses. 1475.