Next Article in Journal
Panoramic Ultrasound Examination of Posterior Neck Extensors in Healthy Subjects: Intra-Examiner Reliability Study
Next Article in Special Issue
Performance of Targeted Library Preparation Solutions for SARS-CoV-2 Whole Genome Analysis
Previous Article in Journal
Long Term Follow-Up of Polish Patients with Isovaleric Aciduria. Clinical and Molecular Delineation of Isovaleric Aciduria
Previous Article in Special Issue
Sensitivity, Specificity and Predictive Values of Molecular and Serological Tests for COVID-19: A Longitudinal Study in Emergency Room
Open AccessArticle

A Rapid SARS-CoV-2 RT-PCR Assay for Low Resource Settings

1
AI Biosciences, Inc., College Station, TX 77845, USA
2
The Medical College of Wisconsin, Milwaukee, WI 53226, USA
*
Author to whom correspondence should be addressed.
Diagnostics 2020, 10(10), 739; https://doi.org/10.3390/diagnostics10100739
Received: 22 August 2020 / Revised: 18 September 2020 / Accepted: 23 September 2020 / Published: 24 September 2020
Quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay is the gold standard recommended to test for acute SARS-CoV-2 infection. However, it generally requires expensive equipment such as RNA isolation instruments and real-time PCR thermal cyclers. As a pandemic, COVID-19 has spread indiscriminately, and many low resource settings and developing countries do not have the means for fast and accurate COVID-19 detection to control the outbreak. Additionally, long assay times, in part caused by slow sample preparation steps, have created a large backlog when testing patient samples suspected of COVID-19. With many PCR-based molecular assays including an extraction step, this can take a significant amount of time and labor, especially if the extraction is performed manually. Using COVID-19 clinical specimens, we have collected evidence that the RT-qPCR assay can feasibly be performed directly on patient sample material in virus transport medium (VTM) without an RNA extraction step, while still producing sensitive test results. If RNA extraction steps can be omitted without significantly affecting clinical sensitivity, the turn-around time of COVID-19 tests, and the backlog we currently experience can be reduced drastically. Furthermore, our data suggest that rapid RT-PCR can be implemented for sensitive and specific molecular diagnosis of COVID-19 in locations where sophisticated laboratory instruments are not available. Our USD 300 set up achieved rapid RT-PCR using thin-walled PCR tubes and a water bath setup using sous vide immersion heaters, a Raspberry Pi computer, and a single servo motor that can process up to 96 samples at a time. Using COVID-19 positive clinical specimens, we demonstrated that RT-PCR assays can be performed in as little as 12 min using untreated samples, heat-inactivated samples, or extracted RNA templates with our low-cost water bath setup. These findings can help rapid COVID-19 testing to become more accessible and attainable across the globe. View Full-Text
Keywords: SARS-CoV-2 virus; RNA; rapid RT-PCR assay; low resource setting; thermal cycler SARS-CoV-2 virus; RNA; rapid RT-PCR assay; low resource setting; thermal cycler
Show Figures

Graphical abstract

MDPI and ACS Style

Arumugam, A.; Faron, M.L.; Yu, P.; Markham, C.; Wu, M.; Wong, S. A Rapid SARS-CoV-2 RT-PCR Assay for Low Resource Settings. Diagnostics 2020, 10, 739.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop