Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Feb 16;16(2):e0246302.
doi: 10.1371/journal.pone.0246302. eCollection 2021.

Evaluation of two automated low-cost RNA extraction protocols for SARS-CoV-2 detection

Fernando Lázaro-Perona  1 , Carlos Rodriguez-Antolín  1 , Marina Alguacil-Guillén  1 , Almudena Gutiérrez-Arroyo  1 , Jesús Mingorance  1 , Julio García-Rodriguez  1 ; SARS-CoV-2 Working Group
Affiliations

Evaluation of two automated low-cost RNA extraction protocols for SARS-CoV-2 detection

Fernando Lázaro-Perona et al. PLoS One. .

Abstract

Background: Two automatable in-house protocols for high-troughput RNA extraction from nasopharyngeal swabs for SARS-CoV-2 detection have been evaluated.

Methods: One hundred forty one SARS-CoV-2 positive samples were collected during a period of 10-days. In-house protocols were based on extraction with magnetic beads and designed to be used with either the Opentrons OT-2 (OT-2in-house) liquid handling robot or the MagMAXTM Express-96 system (MMin-house). Both protocols were tested in parallel with a commercial kit that uses the MagMAXTM system (MMkit). Nucleic acid extraction efficiencies were calculated from a SARS-CoV-2 DNA positive control.

Results: No significant differences were found between both in-house protocols and the commercial kit in their performance to detect positive samples. The MMkit was the most efficient although the MMin-house presented, in average, lower Cts than the other two. In-house protocols allowed to save between 350€ and 400€ for every 96 extracted samples compared to the commercial kit.

Conclusion: The protocols described harness the use of easily available reagents and an open-source liquid handling system and are suitable for SARS-CoV-2 detection in high throughput facilities.

PubMed Disclaimer

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Percentages of SARS-CoV-2 samples positive for the orf1ab, S and N targets with the MMkit, OT-2in-house and MMin-house methods.
The numbers over the bars indicate the percentage of positive samples for each target. Overall, 114 (81%) samples were positive with MMkit, 111 (79%) with OT-2in-house and 118 (84%) with MMin-house.
Fig 2
Fig 2. Boxplot showing the distribution of the Ct values obtained for the orf1ab, S and N targets with the MMkit, OT-2in-house and MMin-house methods.
The y-axis shows the amplification cycle (Ct). The number of data points is shown over each data set.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Merindol N, Pépin G, Marchand C, Rheault M, Peterson C, Poirier A, et al. SARS-CoV-2 detection by direct rRT-PCR without RNA extraction. J Clin Virol. 2020;128: 104423 10.1016/j.jcv.2020.104423 - DOI - PMC - PubMed
    1. Esbin MN, Whitney ON, Chong S, Maurer A, Darzacq X, Tjian R. Overcoming the bottleneck to widespread testing: a rapid review of nucleic acid testing approaches for COVID-19 detection. RNA. 2020;26: 771–783. 10.1261/rna.076232.120 - DOI - PMC - PubMed
    1. Bain W, Lee JS, Watson AM, Stitt‐Fischer MS. Practical Guidelines for Collection, Manipulation and Inactivation of SARS‐CoV‐2 and COVID‐19 Clinical Specimens. Curr Protoc Cytom. 2020;93 10.1002/cpcy.77 - DOI - PMC - PubMed
    1. Wang Y, Song W, Zhao Z, Chen P, Liu J, Li C. The impacts of viral inactivating methods on quantitative RT-PCR for COVID-19. Virus Res. 2020;285: 197988 10.1016/j.virusres.2020.197988 - DOI - PMC - PubMed
    1. Ambrosi C, Prezioso C, Checconi P, Scribano D, Sarshar M, Capannari M, et al. SARS-CoV-2: Comparative analysis of different RNA extraction methods. J Virol Methods. 2021;287: 114008 10.1016/j.jviromet.2020.114008 - DOI - PMC - PubMed

Publication types

MeSH terms