Preparation of norovirus GII loop mediated isothermal amplification freeze-drying microsphere reagents and its application in an on-site integrated rapid detection platform

Norovirus is an infectious disease that can cause non-bacterial gastroenteritis,which has a low infectious dose,rapid onset,and strong transmission ability;therefore,rapid and sensitive detection is essential to reduce the transmission of gastroenteritis.In the study,a norovirus GII loop-mediated isothermal amplification assay was developed and prepared into freeze-drying microspheres,and a closed-cassette-based,integrated,reagent-ambient storage,on-site instant detection platform for norovirus GII was constructed using a commercial,fully automated nucleic acid analyzer with integrated magnetic bearing based nuclear acid extraction and nucleic acid detection,with a sensitivity of 10 copies/μL,with no cross-reactivity with other 5 viruses.For 28 simulated samples,the integrated assay platform was consistent with the experimental results of reverse transcription-quantitative polymerase chain reaction(RT-qPCR)assays after conventional laboratory nucleic acid extraction.The entire process can be finished in about 1 h,which is ideal for immediate rapid detection.