Steps Towards a Pre-Amplification Free Multiplexed Cas13-Based Viral Diagnostic Test
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Mosur, Varun
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Abstract
The recent COVID-19 pandemic has clarified the need for rapid, low-resource point-of-care diagnostic viral tests. Current approaches have a number of shortcomings, including the need for laboratory instrumentation and for trained personnel. We present the first steps of development towards a novel Clustered Regularly Interspaced Short Palindromic Repeats (CRIPSR) (CRISPR-associated protein) Cas13-based diagnostic test that eliminates the need for laboratory instrumentation and any RNA target amplification step while allowing for flexibility and adaptability for future pathogens. Cas13a is complexed with a CRISPR RNA (crRNA) and allowed to detect and cleave target viral RNA. We utilize the secondary trans-cleavage function of the enzyme to cleave a reporter from a surface (a bead), allowing for the reporter to then be separated and transported into a lateral flow-based diagnostic test. This system allows us to separate the detection and readout steps and optimize both independently, as described in the following body of work.
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2023-08-10
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