I am currently developing RPAs for the detection of flaviviruses, however I am struggling a bit with determining the minimum detection limits of my assays. I performed ten-fold dilutions on transcribed RNA (Starting concentration of 8.24 ng/microliter), and performed RPA for dilution 104 , 103, 102 and 101. I followed the guidelines given in the package insert of the PCRD detector sticks, and I was able to only detect 104 (6 microliter amplicon with 84 microliter buffer). According to other articles, my assay is suppose to detect at least till 102. What is a suitable dilution for the lower concentrations?

Any help would be much appreciated!