- Title Information
- Title
- Developing a Predictive Diagnostic for COVID-19
- Type of Resource (primo)
- dissertations
- Name:
Personal
- Name Part
- Lu, Christine
- Role
- Role Term:
Text
- creator
- Name:
Personal
- Name Part
- Nau, Gerard
- Role
- Role Term:
Text
- Advisor
- Name:
Personal
- Name Part
- Monaghan, Sean
- Role
- Role Term:
Text
- Reader
- Name:
Personal
- Name Part
- Fredericks, Alger
- Role
- Role Term:
Text
- Reader
- Name:
Corporate
- Name Part
- Brown University. Biology and Medicine: Biotechnology
- Role
- Role Term:
Text
- sponsor
- Origin Information
- Copyright Date
- 2023
- Physical Description
- Extent
- 5, 30 p.
- digitalOrigin
- born digital
- Note:
thesis
- Thesis (Sc. M.)--Brown University, 2023
- Genre (aat)
- theses
- 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.
- Subject (fast)
(authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/01984643")
- Topic
- COVID-19 (Disease)
- Subject
- Topic
- RT-qPCR
- Language
- Language Term (ISO639-2B)
- English
- Record Information
- Record Content Source (marcorg)
- RPB
- Record Creation Date
(encoding="iso8601")
- 20230602