CRISPR/Cas12-mediated detection of GI and GII Norovirus in different food samples

Abstract

Norovirus is one of the leading causes of infectious diarrhea, occurring in about 18% of diarrhea cases worldwide. Norovirus is characterized by a low infectious dose, rapid onset, and strong transmission capacity. Given the lack of specific drugs and vaccines, developing efficient and accurate detection technologies is of great significance to prevent and control the spread of diseases. This study combined the reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology with the clustered regularly interspaced short palindromic repeats (CRISPR) technology to develop a sensitive and rapid detection method, which can reduce the reliance on temperature control and expensive real-time fluorescent polymerase chain reaction (PCR) devices. The RT-LAMP/CRISPR Cas12a method demonstrated good specificity and sensitivity, testing food samples of three different substrates with 100% positive qualitative accuracy. The detection sensitivity is 32.8 copies/reaction for Norovirus GI and 22.8 copies/reaction for Norovirus GII. This method helps to effectively identify food products contaminated with Norovirus, thereby preventing human infections and economic losses due to disease outbreaks.