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Efficient SARS-CoV-2 Quantitative Reverse Transcriptase PCR Saliva Diagnostic Strategy utilizing Open-Source Pipetting Robots
Published on: February 11, 2022
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A Rapid SARS-CoV-2 RT-PCR Assay for Low Resource Settings
Arunkumar Arumugam1, Matthew L Faron2, Peter Yu1
1AI Biosciences, Inc., College Station, TX 77845, USA.
Diagnostics (Basel, Switzerland)
|September 29, 2020
Summary
A new method allows for rapid COVID-19 testing using quantitative reverse transcription polymerase chain reaction (RT-qPCR) without RNA extraction. This low-cost approach significantly reduces testing time and backlog, making accurate SARS-CoV-2 detection more accessible globally.
Area of Science:
- Molecular Biology
- Infectious Disease Diagnostics
- Biomedical Engineering
Background:
- Quantitative reverse transcription polymerase chain reaction (RT-qPCR) is the gold standard for SARS-CoV-2 detection but requires expensive equipment and lengthy sample preparation.
- The COVID-19 pandemic highlighted the need for rapid, accessible diagnostic tools, especially in resource-limited settings.
- Current RT-qPCR protocols often include a time-consuming RNA extraction step, contributing to testing backlogs.
Purpose of the Study:
- To develop and validate a simplified, low-cost RT-qPCR assay for SARS-CoV-2 detection that omits the RNA extraction step.
- To assess the feasibility of performing RT-qPCR directly on clinical samples in virus transport medium (VTM).
- To demonstrate the potential for rapid molecular diagnosis of COVID-19 in settings lacking sophisticated laboratory infrastructure.
Main Methods:
- A low-cost RT-qPCR setup was created for under USD 300, utilizing a water bath, immersion heaters, a Raspberry Pi, and a servo motor for processing up to 96 samples.
- The assay was performed directly on patient samples in VTM without an RNA extraction step, as well as on heat-inactivated and extracted RNA samples.
- Testing was conducted using COVID-19 positive clinical specimens.
Main Results:
- The simplified RT-qPCR assay achieved sensitive and specific detection of SARS-CoV-2.
- RT-qPCR assays were successfully performed in as little as 12 minutes using the low-cost water bath setup.
- Omitting the RNA extraction step did not significantly compromise clinical sensitivity.
Conclusions:
- Direct RT-qPCR from patient samples without RNA extraction is a feasible and sensitive method for SARS-CoV-2 detection.
- This simplified, low-cost approach can drastically reduce COVID-19 testing turnaround times and alleviate backlogs.
- The developed method offers a promising solution for accessible and rapid COVID-19 diagnosis in resource-limited areas worldwide.
