Journal
LAB ON A CHIP
Volume 22, Issue 18, Pages 3436-3452Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d2lc00434h
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Funding
- Jiangsu Provincial Special Program of Medical Science, China [BE2019749]
- National Natural Science Foundation of China [62071119]
- Jiangsu Provincial Key Research and Development Program, China [BA2020016]
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The COVID-19 pandemic has posed an unprecedented threat to the global health system, especially in resource-limited areas. This study presents a gravity-driven microfluidic cartridge design for rapid and sensitive viral RNA/DNA diagnostic testing without the need for auxiliary hardware. The system exhibited comparable detection performance to the gold standard qPCR instrument and showed high diagnostic accuracy, making it highly promising for infectious disease prevention and control in poor-resource areas.
The pandemic of coronavirus disease 2019 (COVID-19), due to the novel coronavirus (SARS-CoV-2), has created an unprecedented threat to the global health system, especially in resource-limited areas. This challenge shines a spotlight on the urgent need for a point-of-care (POC) quantitative real-time PCR (qPCR) test for sensitive and rapid diagnosis of viral infections. In a POC system, a closed, single-use, microfluidic cartridge is commonly utilized for integration of nucleic acid preparation, PCR amplification and florescence detection. But, most current cartridge systems often involve complicated nucleic acid extraction via active pumping that relies on cumbersome external hardware, causing increases in system complexity and cost. In this work, we demonstrate a gravity-driven cartridge design for an integrated viral RNA/DNA diagnostic test that does not require auxiliary hardware for fluid pumping due to adopted extraction-free amplification. This microfluidic cartridge only contains two reaction chambers for nucleic acid lysis and amplification respectively, enabling a fast qPCR test in less than 30 min. This gravity-driven pumping strategy can help simplify and minimize the microfluidic cartridge, thus enabling high-throughput (up to 12 test cartridges per test) molecular detection via a small cartridge readout system. Thus, this work addresses the scalability limitation of POC molecular testing and can be run in any settings. We verified the analytical sensitivity and specificity of the cartridge testing for respiratory pathogens and sexually transmitted diseases using SARS-CoV-2, influenza A/B RNA samples, and human papillomavirus 16/18 DNA samples. Our cartridge system exhibited a comparable detection performance to the current gold standard qPCR instrument ABI 7500. Moreover, our system showed very high diagnostic accuracy for viral RNA/DNA detection that was well validated by ROC curve analysis. The sample-to-answer molecular testing system reported in this work has the advantages of simplicity, rapidity, and low cost, making it highly promising for prevention and control of infectious diseases in poor-resource areas.
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