A CRISPR/Cas12a-based reverse transcription multiple cross displacement amplification technique for sensitive, rapid, and highly specific detection of human enterovirus A71 in clinical application

Enterovirus A71 (EVA71) is a major pathogen responsible for hand, foot, and mouth disease (HFMD) outbreaks and poses a considerable threat to public health. In this work, we established a molecular detection system that integrates reverse transcription multiple cross displacement amplification (RT-MCDA) with CRISPR/Cas12a technology, targeting the VP1 gene region for EVA71 identification. This platform, termed EVA71-RT-MCDA-CRISPR, utilizes RT-MCDA to achieve efficient pre-amplification of the t