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Title: Accessible Platform for High-Throughput COVID-19 Molecular Diagnostics and Genome Sequencing Using a Repurposed 3D Printer for RNA Extraction.
Austin Authors: Vandelannoote, Koen;Buultjens, Andrew H;Li, Lucy;Sharkey, Liam K;Herisse, Marion;Pidot, Sacha J;Hoang, Tuyet;Howden, Benjamin P ;Monk, Ian R;Seemann, Torsten;Lee, Jean Y H;Stinear, Timothy P
Affiliation: Department of Microbiology and Immunology, Doherty Institute for Infection and Immunity, University of Melbourne, 792 Elizabeth St, Melbourne, Victoria 3000, Australia
Infectious Diseases
Department of Infectious Diseases, Monash Health, 246 Clayton Rd, Clayton, Victoria 3168, Australia
Microbiological Diagnostic Unit Public Health Laboratory, Doherty Institute for Infection and Immunity, University of Melbourne, 792 Elizabeth St, Melbourne, Victoria 3000, Australia
Issue Date: 13-Sep-2021
Date: 2021-08-26
Publication information: ACS Biomaterials Science & Engineering 2021; 7(9): 4669-4676
Abstract: 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.
DOI: 10.1021/acsbiomaterials.1c00775
ORCID: 0000-0002-5984-1328
Journal: ACS Biomaterials Science & Engineering
PubMed URL: 34437802
Type: Journal Article
Subjects: 3D printer
RNA extraction and purification
genome sequencing
molecular diagnostics
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