An immobilization-based, loop-mediated isothermal amplification device for nucleic acid detection of SARS-CoV-2 N gene

Abstract

Abstract Confronting the global spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), simple, fast and specific non-laboratory SARS-CoV-2 diagnostic tests are urgently required. However, the current nucleic acid assays generally rely on the diagnostic laboratory, trained staff and specialized equipment for execution and analysis, presenting clear limitations in the field detection. Here, we describe a portable and reliable immobilization-based loop-mediated isothermal amplification (LAMP) device which is mobile, without the requirement of any complicated instrument and appropriate for high-throughput testing. This device was constructed by utilizing the interaction between a carboxyl-tagged primer and an amino-tagged substrate, and capable of catching the target sequence in SARS-CoV-2 produced via the immobilization-based LAMP. In this study, the immobilization conditions and immobilized primer structure were explored and optimized. With this proposed device, the analysis result can be obtained rapidly in 30 min with excellent specificity, even if the template is extracted from a complex sample containing pharyngeal swab or human blood. In addition, the device can be applied to detect the nucleic acid of SARS-CoV-2 and various other pathogens, showing attractive potential for rapid and high-throughput detection at airports, railway stations, cold-chain transportations, community hospitals and so on. Therefore, we believe that the immobilization-based LAMP device is an advanced approach to developing a portable, specific, low-cost and high-throughput diagnostic platform.