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An Optimal Transport Medium for SARS-CoV-2 Detection in the Direct Method of Rapid Microfluidic PCR System
Miyako Takata1, Masaki Nakamoto2, Tsuyoshi Kitaura2
1Department of Pathobiological Science and Technology, Graduate School of Medical Science, School of Health Science, Faculty of Medicine, Tottori University, Yonago 683-8503, Japan.
Yonago Acta Medica
|November 25, 2024
Summary
The direct method combined with GeneSoC® RT-PCR shows high sensitivity for detecting SARS-CoV-2. eSwab® transport medium is optimal for this rapid point-of-care testing, outperforming VTM.
Area of Science:
- Molecular Biology
- Virology
- Point-of-Care Diagnostics
Background:
- Rapid real-time reverse transcription PCR (RT-PCR) with microfluidic technology is suitable for point-of-care (POC) testing of SARS-CoV-2.
- Direct RNA extraction methods simplify RT-PCR by eliminating purification steps, crucial for rapid testing.
- The choice of transport medium is critical as it can affect subsequent PCR steps, yet its influence on direct methods remains understudied.
Purpose of the Study:
- To evaluate the impact of different transport media on the performance of a direct RNA extraction method combined with rapid microfluidic RT-PCR (GeneSoC®).
- To assess the concordance between direct RNA extraction and traditional RNA purification methods using various transport media.
- To determine the analytical sensitivity and detectability of SARS-CoV-2 RNA using the direct method with different sample transport media.
Main Methods:
- Clinical samples collected in viral transport medium (VTM) and eSwab® were used to evaluate the concordance of direct RNA extraction versus standard RNA purification.
- Spiked samples in generic (H2O, PBS) and commercial transport media (VTM, eSwab®) assessed the sensitivity of GeneSoC® RT-PCR with the direct method.
- Analytical sensitivity and PCR inhibition were evaluated using clinical specimens and diluted samples.
Main Results:
- eSwab® demonstrated higher sensitivity, detecting as low as 1 copy/reaction in spiked samples and <10 copies/reaction in clinical specimens.
- Viral RNA detection was unstable in VTM with <100 copies/reaction, while eSwab® allowed detection below 10 copies/reaction.
- Concordance between direct method and RNA extraction was significantly higher with eSwab® (85%) compared to VTM (38%).
Conclusions:
- The direct method combined with GeneSoC® RT-PCR demonstrates high sensitivity for SARS-CoV-2 detection.
- eSwab® is a suitable transport medium for direct RNA extraction coupled with rapid microfluidic RT-PCR, yielding reliable results.
- This combination offers a promising approach for rapid and efficient point-of-care SARS-CoV-2 testing.
