ABSTRACT
Although real‐time quantitative reverse transcription polymerase chain reaction (RT‐qPCR) is the gold standard for detecting the virus severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) and other pathogens, the coronavirus disease 2019 (COVID‐19) pandemic has highlighted the scarcity of instruments, devices, and reagents for polymerase chain reaction (PCR) testing in constrained settings. At least for under‐resourced countries, it has become critical to deploy instruments that can be rapidly constructed and satisfy this demand. Instead of separating the optical system from the thermal module (typical of qPCR thermocyclers), we report a portable Hybrid Opto‐Thermocycler—dubbed HybOT Cycler—that takes advantage of the high‐temperature tolerances (>100 °C) of electronic and optical components to combine thermal control, illumination, and fluorescence detection into a highly integrated hybrid module. This simple configuration allowed us to reduce the overall number of components, thus simplifying its assembly and reducing the instrument size. The HybOT Cycler is wirelessly controlled from an application installed in a tablet. PCR assays are carried out in a bubble‐free microfluidic device that can be easily replicated from an acrylic mold. Using the HybOT Cycler, down to 100 copies/µL of genetic material of the virus SARS‐CoV‐2 with 95% sensitivity and 100% specificity is detected. The HybOT Cycler can assist in diagnosing SARS‐CoV‐2 and other pathogens in resource‐poor settings. [ FROM AUTHOR] Copyright of Advanced Materials Technologies is the property of John Wiley & Sons, Inc. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full . (Copyright applies to all s.)
ABSTRACT
Although real‐time quantitative reverse transcription polymerase chain reaction (RT‐qPCR) is the gold standard for detecting the virus severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) and other pathogens, the coronavirus disease 2019 (COVID‐19) pandemic has highlighted the scarcity of instruments, devices, and reagents for polymerase chain reaction (PCR) testing in constrained settings. At least for under‐resourced countries, it has become critical to deploy instruments that can be rapidly constructed and satisfy this demand. Instead of separating the optical system from the thermal module (typical of qPCR thermocyclers), we report a portable Hybrid Opto‐Thermocycler—dubbed HybOT Cycler—that takes advantage of the high‐temperature tolerances (>100 °C) of electronic and optical components to combine thermal control, illumination, and fluorescence detection into a highly integrated hybrid module. This simple configuration allowed us to reduce the overall number of components, thus simplifying its assembly and reducing the instrument size. The HybOT Cycler is wirelessly controlled from an application installed in a tablet. PCR assays are carried out in a bubble‐free microfluidic device that can be easily replicated from an acrylic mold. Using the HybOT Cycler, down to 100 copies/µL of genetic material of the virus SARS‐CoV‐2 with 95% sensitivity and 100% specificity is detected. The HybOT Cycler can assist in diagnosing SARS‐CoV‐2 and other pathogens in resource‐poor settings. [ FROM AUTHOR]
ABSTRACT
The applications of serology tests to the virus SARS-CoV-2 are diverse, ranging from diagnosing COVID-19, understanding the humoral response to this disease, and estimating its prevalence in a population, to modeling the course of the pandemic. COVID-19 serology assays will significantly benefit from sensitive and reliable technologies that can process dozens of samples in parallel, thus reducing costs and time; however, they will also benefit from biosensors that can assess antibody reactivities to multiple SARS-CoV-2 antigens. Here, we report a high-throughput microfluidic device that can assess antibody reactivities against four SARS-CoV-2 antigens from up to 50 serum samples in parallel. This semi-automatic platform measures IgG and IgM levels against four SARS-CoV-2 proteins: the spike protein (S), the S1 subunit (S1), the receptor-binding domain (RBD), and the nucleocapsid (N). After assay optimization, we evaluated sera from infected individuals with COVID-19 and a cohort of archival samples from 2018. The assay achieved a sensitivity of 95% and a specificity of 91%. Nonetheless, both parameters increased to 100% when evaluating sera from individuals in the third week after symptom onset. To further assess our platform's utility, we monitored the antibody titers from 5 COVID-19 patients over a time course of several weeks. Our platform can aid in global efforts to control and understand COVID-19.