Accessible Platform for High-Throughput COVID-19 Molecular Diagnostics and Genome Sequencing Using a Repurposed 3D Printer for RNA Extraction
| Autor: | Marion Herisse, Andrew H. Buultjens, Tuyet Hoang, Benjamin P Howden, Ian R. Monk, Timothy P. Stinear, Liam K. R. Sharkey, Jean Y. H. Lee, Sacha J. Pidot, Koen Vandelannoote, Lucy Li, Torsten Seemann |
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| Rok vydání: | 2021 |
| Předmět: |
Consumables
Coronavirus disease 2019 (COVID-19) RNA extraction and purification Computer science RT-PCR Biomedical Engineering Article DNA sequencing 3d printer molecular diagnostics Biomaterials Humans 3D printer Pathology Molecular Pandemics Throughput (business) SARS-CoV-2 business.industry COVID-19 Molecular diagnostics genome sequencing Nasal Swab Embedded system RNA Viral RNA extraction business |
| Zdroj: | ACS Biomaterials Science & Engineering |
| ISSN: | 2373-9878 |
| Popis: | The COVID-19 pandemic has exposed the dependence of diagnostic laboratories on a handful of large corporations with market monopolies on the worldwide supply of reagents, consumables, and hardware for molecular diagnostics. Global shortages of key consumables for RT-qPCR detection of SARS-CoV-2 RNA have impaired the ability to run essential, routine diagnostic services. Here, we describe a workflow for rapid detection of SARS-CoV-2 RNA in upper respiratory samples including nasal swabs and saliva, utilizing low-cost equipment and readily accessible reagents. Using repurposed Creality3D Ender-3 three-dimensional (3D) printers, we built a semiautomated paramagnetic bead RNA extraction platform. The hardware for the system was built for $300 USD, and the material cost per reaction was $1 USD. Named the Ender VX500, instrument performance when paired with RT-qPCR for SARS-CoV-2 detection in nasal and saliva specimens was two virus copies per microliter. There was a high-performance agreement (assessed using 458 COVID-19 nasal swab specimens) with the Aptima SARS-CoV-2 assay run on the Hologic Panther, a commercial automated RNA extraction and detection platform. Inter- and intrainstrument precision was excellent (coefficients of variation (CoV) of 1.10 and 0.66–1.32%, respectively) across four instruments. The platform is scalable with throughput ranging from 23 specimens on a single instrument run by one user in 50 min to 364 specimens on four instruments run by four users in 190 min. Step-by-step instructions and protocols for building and running the Ender VX500 have been made available without restriction. |
| Databáze: | OpenAIRE |
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