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Updated: Oct 2, 2025

Efficient SARS-CoV-2 Quantitative Reverse Transcriptase PCR Saliva Diagnostic Strategy utilizing Open-Source Pipetting Robots
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Efficient SARS-CoV-2 Quantitative Reverse Transcriptase PCR Saliva Diagnostic Strategy utilizing Open-Source

Rachel E Ham1, Austin R Smothers2, Kylie L King1

  • 1Center for Innovative Medical Devices and Sensors (REDDI Lab), Clemson University.

Journal of Visualized Experiments : Jove
|February 28, 2022
PubMed
Summary
This summary is machine-generated.

This study presents an automated saliva-based diagnostic assay for COVID-19 surveillance. The method offers rapid, accurate detection and cost-effective testing for large populations.

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Area of Science:

  • Epidemiology
  • Molecular Diagnostics
  • Public Health

Background:

  • The COVID-19 pandemic highlighted challenges in rapid diagnostic testing, especially in congregate settings.
  • Limited supply chains for traditional nucleic acid tests, like nasopharyngeal swabs, caused delays.
  • Saliva-based testing offers a less invasive alternative with comparable accuracy to swab-based methods.

Purpose of the Study:

  • To develop and implement an automated multiplex RT-qPCR assay for SARS-CoV-2 surveillance.
  • To enable efficient and accurate COVID-19 testing in a university and surrounding community setting.
  • To optimize workflow and system flexibility using open-source automation.

Main Methods:

  • Development of a multiplex RT-qPCR assay using saliva samples.
  • Implementation of open-source liquid handling robots and thermocyclers for automation.
  • Validation of the automated system for detecting a wide range of viral RNA concentrations.

Main Results:

  • The automated saliva-based RT-qPCR assay provided rapid results, with 95% of samples processed in under 9 hours and 99% in under 24 hours.
  • The cost per test was significantly reduced to $2.80 with bulk reagent purchasing.
  • The system demonstrated accuracy comparable to existing methods for population surveillance.

Conclusions:

  • Automated saliva-based RT-qPCR is a viable strategy for efficient and scalable COVID-19 population surveillance.
  • This approach offers a flexible, cost-effective, and rapid diagnostic solution for public health.
  • The developed assay can meet both large- and small-scale testing demands effectively.