An integrated platform for rotavirus nucleic acid detection

Abstract

Group A rotavirus (RVA) is the main cause of acute gastroenteritis in infants and children with high infectivity, continuously presenting a negative impact on society. Therefore, the rapid and accurate detection of RVA is crucial for the prevention and diagnosis of the disease. Here we establish a modular and visual diagnostic platform for nucleic acid detection of the pathogen by integrating four technologies, asymmetric nucleic acid sequence-based amplification (asymmetric-NASBA), enzyme-free toehold-mediated strand displacement (TMSD), programmable primer exchange reaction (PER) cascades-mediated efficient signal amplification and a lateral flow immunoassay (LFIA), namely an ATPL-based platform. The built-in workflow can isothermally complete a convenient and rapid detection of the target molecules with a minimum concentration of 2.46 copies/mL in about 20 min, and markedly generate a lateral flow line signal for the naked eye. Moreover, a single-base difference can be distinguished from the target sequence, demonstrating high specificity and clinically analytical performance. Compared to the current common detection technology RT-qPCR, this platform provides detection results with high consistency with a kappa value of 0.927. In a word, the established ATPL-based detection platform is fast, highly sensitive and specific with promising accuracy, suggesting wide application potential in the detection of rotavirus or other nucleic acid in the future.