Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays
Figure 5
RT-QuIC detection and end point dilution analysis of nasal lavages from HY-TME-infected hamsters.
At the clinical stage of disease in hamsters i.c.-inoculated with HY-TME, nasal lavage samples (∼1 ml) were collected from individual hamsters, serially diluted and used to seed RT-QuIC reactions. (A) Detection of HY-TME in nasal lavages is shown. As positive and negative controls, 5×10-6-fold dilutions of hamster scrapie BH (containing ∼100 fg PrPSc) and NBH, respectively, were used as seeds. Dilutions of HY-TME brain were also tested for comparison. The data points show the average ThT fluorescence of 8 replicate wells. (B) Three representative HY-TME infected hamster lavages were serially diluted and analyzed by RT-QuIC. Each data point represents the percentage of replicate wells (n = 4) that are ThT-positive according to the criteria in Material and Methods as a function of nasal lavage dilution factor. The tabular data from lavages tested here can be found in Table 2.