Abstract
The coronavirus disease 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus (SARS-CoV)-2, is rapidly spreading and severely straining the capacities of public health communities and systems around the world. Therefore, accurate, rapid, and robust diagnostic tests for COVID-19 are crucial to prevent further spread of the infection, alleviate the burden on healthcare and diagnostic facilities, and ensure timely therapeutic intervention. To date, several detection methods based on nucleic acid amplification have been developed for the rapid and accurate detection of SARS-CoV-2. Despite the myriad of advancements in the detection methods for SARS-CoV-2, rapid sample preparation methods for RNA extraction from viruses have rarely been explored. Here, we report a rapid COVID-19 molecular diagnostic system that combines a self-powered sample preparation assay and loop-mediated isothermal amplification (LAMP) based naked-eye detection method for the rapid and sensitive detection of SARS-CoV-2. The self-powered sample preparation assay with a hydrophilic polyvinylidene fluoride filter and dimethyl pimelimidate can be operated by hand, without the use of any sophisticated instrumentation, similar to the reverse transcription (RT)-LAMP-based lateral flow assay for the naked-eye detection of SARS-CoV-2. The COVID-19 molecular diagnostic system enriches the virus population, extracts and amplifies the target RNA, and detects SARS-CoV-2 within 60 min. We validated the accuracy of the system by using 23 clinical nasopharyngeal specimens. We envision that this proposed system will enable simple, facile, efficient, and inexpensive diagnosis of COVID-19 at home and the clinic as a pre-screening platform to reduce the burden on the medical staff in this pandemic era.
Highlights
The pandemic of coronavirus disease 2019 (COVID-19) has emerged as a life-threatening respiratory disease caused by the severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) [1,2]
dimethyl pimelimidate (DMP) consists of methylene groups and bifunctional imidoester groups
The positively charged DMP, whose charge results from the amino group, directly captured the negatively charged virus via electrostatic adsorption (Step 1)
Summary
The pandemic of coronavirus disease 2019 (COVID-19) has emerged as a life-threatening respiratory disease caused by the severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) [1,2]. Traditional pathogen sample preparation methods include enrichment and extraction, which are widely used in clinical diagnosis, and which are based on density gradient centrifugation and culture [45,46] These methods pose various constraints, such as contamination and pathogen spread and loss, the requirement of sophisticated instrumentation and expensive reagents, and a laborintensive process [47,48]. We report a rapid COVID-19 molecular diagnostic system combining a simple virus enrichment and extraction method that involves dimethyl pimelimidate (DMP), a hydrophilic polyvinylidene fluoride (PVDF) filter, and a LAMP-LFA method This proposed system could provide a sensitive, facile, efficient, and inexpensive method for the diagnosis of COVID-19. The proposed COVID-19 diagnostic system is suitable for rapid, facile, and simple diagnosis at inadequately equipped or under-staffed healthcare centers and in situations where results are urgently required for the optimal diagnosis of emerging infectious diseases
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