Effective containment of the COVID-19 pandemic requires rapid and accurate detection of the pathogen.Polymerase chain reaction(PCR)remains the gold standard for COVID-19 confirmation.In this article,we report the perf...Effective containment of the COVID-19 pandemic requires rapid and accurate detection of the pathogen.Polymerase chain reaction(PCR)remains the gold standard for COVID-19 confirmation.In this article,we report the performance of a cost-effective modular microfluidic reverse transcription(RT)-PCR and RT-loop mediated isothermal amplification(RTLAMP)platform,Epidax®,for the point-of-care testing and confirmation of SARS-CoV-2.This platform is versatile and can be reconfigured either for screening using endpoint RT-PCR or RT-LAMP tests or for confirmatory tests using realtime RT-PCR.Epidax®is highly sensitive and detects as little as 1 RNA copy perµL for real-time and endpoint RT-PCR,while using only half of the reagents.We achieved comparable results with those of a commercial platform when detecting SARS-CoV-2 viruses from 81 clinical RNA extracts.Epidax®can also detect SARS-CoV-2 from 44 nasopharyngeal samples without RNA extraction by using a direct RT-PCR assay,which shortens the sample-to-answer time to an hour with minimal user steps.Furthermore,we validated the technology using an RT-LAMP assay on 54 clinical RNA extracts.Overall,our platform provides a sensitive,cost-effective,and accurate diagnostic solution for lowresource settings.展开更多
基金the National University of Singapore(NUS),grant reference number NUSCOVID19RG-34.
文摘Effective containment of the COVID-19 pandemic requires rapid and accurate detection of the pathogen.Polymerase chain reaction(PCR)remains the gold standard for COVID-19 confirmation.In this article,we report the performance of a cost-effective modular microfluidic reverse transcription(RT)-PCR and RT-loop mediated isothermal amplification(RTLAMP)platform,Epidax®,for the point-of-care testing and confirmation of SARS-CoV-2.This platform is versatile and can be reconfigured either for screening using endpoint RT-PCR or RT-LAMP tests or for confirmatory tests using realtime RT-PCR.Epidax®is highly sensitive and detects as little as 1 RNA copy perµL for real-time and endpoint RT-PCR,while using only half of the reagents.We achieved comparable results with those of a commercial platform when detecting SARS-CoV-2 viruses from 81 clinical RNA extracts.Epidax®can also detect SARS-CoV-2 from 44 nasopharyngeal samples without RNA extraction by using a direct RT-PCR assay,which shortens the sample-to-answer time to an hour with minimal user steps.Furthermore,we validated the technology using an RT-LAMP assay on 54 clinical RNA extracts.Overall,our platform provides a sensitive,cost-effective,and accurate diagnostic solution for lowresource settings.
