Development of a Recombinase Polymerase Amplification Assay for Detection of Epidemic Human Noroviruses

Moore, Matthew D.; Jaykus, Lee-Ann

Although several isothermal techniques for the amplification and detection of purified nucleic acids exist 21, 22 , RPA has multiple advantages, i.e., (i.) the use of a single tube; (ii.) real-time detection of amplified product using a fluorescent probe; (iii.) the inclusion of most required reagents in the form of a freeze-dried pellet to simplify testing; (iv.) the inclusion of a binding protein that has been reported to aid in amplification of RNA targets having Table 3 . Exclusivity (specificity) analysis. The genomic RNA/DNA of several enteric viruses and bacteria was extracted using a NucliSens EasyMAG (bioMerieux), and 10 −2 and 10 −3 dilutions of the extracts were loaded as template in the RT-RPA assay using the G2F5-G2R11-G2P1 primer-probe set. c Showed low reactivity in one replicate. a Source organism type used. All organisms may be present in human enteric samples. b Whether or not an amplifiable signal was observed at any point for any dilution with RT-RPA. If yes, then the proportion of replicates for which a positive signal was obtained is presented in parentheses.