Point-of-Care Testing for Norovirus Typing Using CRISPR/Cas12a Combined with Reverse Transcription Recombinase Polymerase Amplification

Noroviruses (NoVs) are one of the leading causes of acutegastroenteritisin humans. This study combined reverse transcription recombinase polymeraseamplification (RT-RPA) with a clustered regularly interspaced shortpalindromic repeat/CRISPR-associated protein (CRISPR/Cas) nucleicacid detection system to develop a point-of-care testing (POCT) technologyfor typing NoVs. The detection can be completed within 35 min at 37 degrees C, covering each genotype of genogroup I (GI) and II (GII) NoVs.The sensitivity of this method is 10 copies/mu L for GI and 1copy/mu L for GII NoV plasmids. For the detection of clinicalsamples, the detection results of this method for NoV infected samplesare consistent with the RT-qPCR detection method in the laboratory,and this detection method has no cross-reactivity with rotavirus andadenovirus. Therefore, the detection method established in this studyenables the diagnosis and screening of suspected patients and closecontacts by POCT, which is important for the timely identificationand control of NoV outbreaks. In addition, the typing detection ofGI and GII NoVs can achieve a precise diagnosis and treatment of patientsinfected with NoVs.