Graphpad Prism 6 Serial Number 575
Graphpad Prism 6 Serial Number 575 - https://urlin.us/2t9JvJ
The detection limit of the method was determined by plotting the Ct values against the log10copy numbers for the eight replicates of each of the dilutions. Regression analysis was performed using Prism 5.0 software (GraphPad, USA). The limit of detection was defined as the dilution at which the proportion of positive wells is equal to 0.5 and the limit of quantification was defined as the dilution at which the proportion of positive wells is equal to 0.67.
The limit of detection was determined by averaging the Ct values for eight replicates of each of the dilutions (7.8 copies/reaction). The results were plotted using Prism 5.0 software (GraphPad, USA). The limit of detection was defined as the dilution at which the proportion of positive wells was equal to 0.5. The limit of quantification was determined by averaging the Ct values for eight replicates of the 10^4^ copies/reaction dilution. The results were plotted using Prism 5.0 software (GraphPad, USA).
The overall assay detection limit was determined to be 75 copies/mL (95% CI, 68.6--81.4 copies/mL) when tested with 10-fold serially diluted virus stocks (inoculum containing 106 copies/mL). In our hands, the efficiency of RPA detection system was found to be 100% using ReaGene detection reagent .
We then analysed ten respiratory specimens that were negative by the SARS-CoV-2 real-time RT-PCR assay, which was performed using the primers and probe recommended by the U.S. Centers for Disease Control and Prevention (CDC) ( Coronavirus Lab Test, Real-Time, FDA Emergency Use Authorization ). The RPA test was performed with the same primers and probes of the CDC kit, and the copy numbers were determined by the same semi-log regression method as described above. The assay was found to be negative in all of the ten respiratory samples tested.
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One of the drawbacks to this approach was that we needed to recompile every time we made even a small change to the code. This slowed everything else we were doing down and because we couldn't always have someone waiting for the application to finish building. 827ec27edc