A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein to Facilitate the Detection of SARS-CoV-2

Abstract

<jats:title>Abstract</jats:title><jats:p>Management of the COVID-19 pandemic requires widespread SARS-CoV-2 testing. A main limitation for widespread SARS-CoV-2 testing is the global shortage of essential supplies, among these, RNA extraction kits. The need for commercial RNA extraction kits places a bottleneck on tests that detect SARS-CoV-2 genetic material, including PCR-based reference tests. Here we propose an alternative method we call PEARL (<jats:underline>P</jats:underline>recipitation <jats:underline>E</jats:underline>nhanced <jats:underline>A</jats:underline>nalyte <jats:underline>R</jats:underline>etrieva<jats:underline>L</jats:underline>) that addresses this limitation. PEARL uses a lysis solution that disrupts cell membranes and viral envelopes while simultaneously providing conditions suitable for alcohol-based precipitation of RNA, DNA, and proteins. PEARL is a fast, low-cost, and simple method that uses common laboratory reagents and offers comparable performance to commercial RNA extraction kits. PEARL offers an alternative method to isolate host and pathogen nucleic acids and proteins to streamline the detection of DNA and RNA viruses, including SARS-CoV-2.</jats:p>

A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic Material

“…Finally, while we designed CREST to be an accessible and scalable assay for detecting SARS-CoV-2, it still requires RNA extraction using commercial kits, which limits its widespread adoption. In a companion paper, we present a method called PEARL (Precipitation Enhanced Analyte RetrievaL) which uses common laboratory reagents to bypass this limitation 29 . To lift the final obstacle to CREST's accessibility, we coupled PEARL to CREST and found that commercial RNA extraction could be omitted (Fig.…”

A Scalable, Easy-to-Deploy Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic Material

“…Finally, while we designed CREST to be an accessible and scalable assay for detecting SARS-CoV-2, it still requires RNA extraction using commercial kits, which limits its widespread adoption. In a companion paper, we present a method called PEARL (Precipitation Enhanced Analyte RetrievaL) which uses common laboratory reagents to bypass this limitation (24). To lift the final obstacle to CREST's accessibility, we coupled PEARL to CREST and found that commercial RNA extraction could be omitted (Fig.…”

“…uploaded to the cloud, enabling distributed point-of-care (POC) testing. Operational costs could be further lowered by multiplexing the amplification of viral targets.Current methods to detect SARS-CoV-2 presence or prior exposure in patient samples rely on detection of the viral genome or antibodies in nucleic acid and serological tests, respectively24 . The detection of viral genome sequences by RT-qPCR is recommended by public health organizations worldwide.…”

The impact of SARS‐CoV‐2 in dementia across Latin America: A call for an urgent regional plan and coordinated response

“…The same group has also reported a field-deployable RNA-extraction procedure called PEARL. 42 Use of the test in Ecuador is currently under way. Similarly, a laboratory in Brazil has developed a SARS-CoV-2 detection method in saliva, using RT-LAMP (loop mediated isothermal amplification), 43 which is also scalable and relatively inexpensive.…”