A RAPID PAPER-BASED COLORIMETRIC MOLECULAR TEST FOR SARS-COV-2 POINT-OF-CARE DIAGNOSTIC
In the year of 2020, an international pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has afflicted tens of millions of people’s life also disrupting global economics. Diagnostic testing is an important part of ensuring public health until a vaccine that has been shown to be safe and effective is made available to the general public. Most tests for detecting COVID-19 utilize quantitative polymerase chain reaction (qPCR) assays, which is a specific and relatively simple quantitative assay that could provide adequate sensitivity for diagnosing early infection. Although powerful, these lab-based molecular assays have a significant lag time, usually several days before receiving results. To satisfy the needs of different purposes (diagnostics, screening, and surveillance), a unified approach is impractical. This thesis presents an alternative testing method supporting the current procedure of point of care (POC) testing and in community testing. This paper-based test overcomes the limitations of current testing methods by utilizing reverse-transcription loop-mediated isothermal amplification (RT-LAMP) and receiving the result on-site by a color change in the presence of the virus within 60 minutes. The test utilizes untreated freshly collected saliva, a less invasive specimen, as the sample and possesses a limit of detection (LoD) of 200 copies of virus per microliter of whole saliva with an analytical sensitivity of 97% and analytical specificity of 100%. The test requires minimal operator training and could be fabricated on a large-scale using roll-to-roll methods. Since the test is based on nucleic acids, the testing platform itself lends to further applications including food safety monitoring, animal diagnostic, etc. simply by changing the specific primers.
- Master of Science in Agricultural and Biological Engineering
- Agricultural and Biological Engineering
- West Lafayette