Author: Moore, Matthew D.; Jaykus, Lee-Ann
Title: Development of a Recombinase Polymerase Amplification Assay for Detection of Epidemic Human Noroviruses Document date: 2017_1_9
ID: 03a6hq3t_5
Snippet: To test the analytical sensitivity (detection limit) of the RT-RPA assay, serial dilutions of purified GII.4 New Orleans RNA from three patients (isolates 29, 47, and 58) were tested for 8 replicates and probit regressions used to interpolate a limit of detection (Fig. 2 ). The range of input RNA producing consistently positive results for all three replicates was 3.2 to 6.2 LGC with time to threshold florescence of 12.4 ± 2.4 min to 8.2 ± 0.5 .....
Document: To test the analytical sensitivity (detection limit) of the RT-RPA assay, serial dilutions of purified GII.4 New Orleans RNA from three patients (isolates 29, 47, and 58) were tested for 8 replicates and probit regressions used to interpolate a limit of detection (Fig. 2 ). The range of input RNA producing consistently positive results for all three replicates was 3.2 to 6.2 LGC with time to threshold florescence of 12.4 ± 2.4 min to 8.2 ± 0.5 min, respectively. Based on the probit regressions average, the limit of detection of the assay (the level of input RNA for which 95% of samples would be positive) is 3.40 ± 0. 20 LGC. This is about 1.0 LGC higher than the limit of detection determined for RT-qPCR, which was predicted by probit to be 2.3 ± 0.04 LGC.
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