Considerations Regarding RNA Extraction Kits for SARS-CoV-2 Detection

Posted on by Jackie Surls
Considerations Regarding RNA Extraction Kits for SARS-CoV-2 Detection

The novel coronavirus disease (COVID-19) pandemic continues to trigger rapid development of molecular diagnostic tests around the world. Nucleic acid amplification-based tests (NAATs)—such as real-time PCR and isothermal amplification—are the gold standard in molecular diagnostic technologies that are being utilized to detect SARS-CoV-2, the COVID-19 disease-causing virus.1

There are two critical and sequential steps that must occur prior to running a SARS-CoV-2 NAAT. The first is proper sample collection per the CDC’s guidelines,2 because inadequate collection may lead to inaccurate or inconclusive diagnostic results. The second is effective viral RNA extraction. The CDC has qualified and validated several extraction systems for use with their 2019-nCoV real-time RT-PCR Diagnostic Panel,3 including systems from Qiagen, Roche, and Biomerieux, but the rapid increase in testing has led to a global shortage of commercially available extraction kits. Consequently, lab test sites need to explore alternative vendors for extraction kits to qualify and validate with their respective NAAT.

Most commercial RNA extraction kits utilize similar principles for RNA extraction, with guanidine thiocyanate being the most popular chaotropic agent to lyse the virus, denature RNases, and protect RNA from degradation. These kits often include additional protocol steps, some designated as optional to improve purity, yield, or analyte detection. When selecting an RNA extraction kit, remember that SARS-CoV-2 is an RNA virus, and thus requires protection from RNases. Understanding the kit reagents and protocol will help ensure quality RNA extraction and reproducible results.

Luminex’s ARIES® SARS-CoV-2 Assay uses a sample-to-answer PCR platform that performs nucleic acid extraction, target amplification, and detection. It is one of the commercially available NAATs on the market.

Jackie Surls, PhD, is a Development & Applications Scientist at Luminex Corporation.

References

  1. “Coronavirus Test Tracker: Commercially Available COVID-19 Diagnostic Tests.” Genome Web (Internet). Cited Mar 2020. Available from: https://www.360dx.com/coronavirus-test-tracker-launched-covid-19-tests.
  2. “Interim Guidelines for Collecting, Handling, and Testing Clinical Specimens from Persons for Coronavirus Disease 2019 (COVID-19).” The Centers for Disease Control and Prevention (Internet). Cited Mar 2020. Available from: https://www.cdc.gov/coronavirus/2019-ncov/lab/guidelines-clinical-specimens.html.
  3. “CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR Diagnostic Panel.” The Centers for Disease Control and Prevention (Internet). Cited Mar 2020. Available from: https://www.fda.gov/media/134922/download.

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