Description
- Abstract:
- Objective: SARS-CoV-2 is an ongoing threat to human health. A predictive diagnostic would be helpful for providing information regarding the future progression of COVID-19 in hospitalized patients, however, none have been deployed. There are polymerase chain reaction (PCR) primers based on the sequences used by the CDC (CDC-N1) for diagnostics from nasopharyngeal swabs, but these primers are not effective for measuring RNAemia in all hospitalized COVID-19 patients. The objective of this project was to use deep RNA sequencing to develop a test with greater sensitivity than that of current diagnostics and provide a lead for developing a predictive diagnostic. Methods: Primers targeting the ORF1ab and N genes were designed and validated through reverse transcription polymerase chain reaction (RT-PCR). The performance of our primers was compared to that of the CDC-N1. RNA was extracted from whole blood of COVID-19 patients admitted to the ICU. Primer designs were evaluated by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results: An optimized RT-qPCR protocol was developed that minimizes background signal and detects SARS-CoV-2 sequences in whole blood. Higher levels of SARS-CoV-2 RNA were found in COVID-19 patients using our primers than with the CDC-N1 primers. Conclusions: We found that our primers are consistently more sensitive than the CDC-N1 primers for the samples tested. Therefore, further studies of our primers should be conducted to elucidate their potential as a predictive diagnostic.
- Notes:
- Thesis (Sc. M.)--Brown University, 2023
Citation
Lu, Christine,
"Developing a Predictive Diagnostic for COVID-19"
(2023).
Biotechnology, Biology and Medicine Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:nwe4e2sr/
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Biology and Medicine Theses and Dissertations
Theses and Dissertations for the Biology and Medicine department....