| Abstrakt: |
Norovirus (NoV) is one of the major pathogens causing acute gastroenteritis diseases worldwide and is highly susceptible to outbreak transmission, increasing medical and economic burden. Objective: To develop a novel detection method for NoV by recombinase aided amplification (RAA). Methods Based on the cDNA sequences corresponding to the GI and Gil NoV detection targets specified in ISO TS 15216-2-2013 and GB 4789.42, the optimal RAA primers and probes were designed and screened, and their specificity for other common food-borne diarrhea viruses was determined; the shortest detection time, reaction procedure, and reaction system were determined by optimization, and the analysis of this detection system was performed. The shortest detection time, reaction procedure and reaction system were optimized, and the sensitivity of the assay system for the detection of NoV reference plasmids and real samples was analyzed, thus establishing a rapid visualization method for the detection of GI and Gil NoV RAA. The optimized reaction procedure can shorten the detection time to about 10 min and reduce the reaction cost by two-thirds, and the sensitivity of the method can reach 10-2 ng/µL for the reference plasmid and 1 ng/µL for the real sample. The two NoV assays established are specific, sensitive, simple, rapid and visualized, and provide a good basis for future rapid NoV detection. [ABSTRACT FROM AUTHOR] |
