Loading...
Please wait, while we are loading the content...
Similar Documents
Microfluidic Nano-Scale qPCR Enables Ultra-Sensitive and Quantitative Detection of SARS-CoV-2
| Content Provider | MDPI |
|---|---|
| Author | Xie, Xin Gjorgjieva, Tamara Attieh, Zaynoun Dieng, Mame Arnoux, Marc Khair, Mostafa Moussa, Yasmine Jallaf, Fatima Al Rahiman, Nabil Jackson, Christopher Messery, Lobna Pamplona, Khristine Victoria, Zyrone Zafar, Mohammed Ali, Raghib Piano, Fabio Gunsalus, Kristin Idaghdour, Youssef |
| Copyright Year | 2020 |
| Description | A major challenge in controlling the COVID-19 pandemic is the high false-negative rate of the commonly used RT-PCR methods for SARS-CoV-2 detection in clinical samples. Accurate detection is particularly challenging in samples with low viral loads that are below the limit of detection (LoD) of standard one- or two-step RT-PCR methods. In this study, we implemented a three-step approach for SARS-CoV-2 detection and quantification that employs reverse transcription, targeted cDNA preamplification, and nano-scale qPCR based on a commercially available microfluidic chip. Using SARS-CoV-2 synthetic RNA and plasmid controls, we demonstrate that the addition of a preamplification step enhances the LoD of this microfluidic RT-qPCR by 1000-fold, enabling detection below 1 copy/µL. We applied this method to analyze 182 clinical NP swab samples previously diagnosed using a standard RT-qPCR protocol (91 positive, 91 negative) and demonstrate reproducible and quantitative detection of SARS-CoV-2 over five orders of magnitude (6 viral copies/µL). Crucially, we detect SARS-CoV-2 with relatively low viral load estimates (<1 to 40 viral copies/µL) in 17 samples with negative clinical diagnosis, indicating a potential false-negative rate of 18.7% by clinical diagnostic procedures. In summary, this three-step nano-scale RT-qPCR method can robustly detect SARS-CoV-2 in samples with relatively low viral loads (<1 viral copy/µL) and has the potential to reduce the false-negative rate of standard RT-PCR-based diagnostic tests for SARS-CoV-2 and other viral infections. |
| Starting Page | 1425 |
| e-ISSN | 22279717 |
| DOI | 10.3390/pr8111425 |
| Journal | Processes |
| Issue Number | 11 |
| Volume Number | 8 |
| Language | English |
| Publisher | MDPI |
| Publisher Date | 2020-11-09 |
| Access Restriction | Open |
| Subject Keyword | Processes Analytical Chemistry Sars-cov-2 Covid-19 Microfluidics Nano-qpcr Ultra-sensitive Viral Rna Viral Load Detection |
| Content Type | Text |
| Resource Type | Article |
