Plasma PfHRP2 measurement via serial dilution showed minimal bias compared to ELISA and acceptable limits of agreement. The Paracheck ® brand showed the strongest correlation (r 2 = 0.963) as well as the lowest inter-observer variability (combined kappa across band intensities for three observers = 0.938). RDTs were read by eye and digital capture, and PfHRP2 concentrations were measured via serial dilution with results compared to ELISA readings. Plasma samples covering a wide range of PfHRP2 concentrations were applied to rapid diagnostic tests (RDTs). You can see to the left that Prism has createda new page in the "Results" folder, labeled "Interpolation of Data1," and this has two subpages.Plasma Plasmodium falciparum histidine-rich protein-2 (PfHRP2) is the most accurate biomarker for severe malaria, but its measurement by ELISA has been considered too unwieldy to incorporate into clinical management. You will also check the "Plot curve"option and leave it set to the 95% confidenceband, so that you can examine how well the curve fits the data. Here, you can tell Prism to calculatea confidence interval for the interpolateddata, but this is optional so you will not do it here. You will leave the Outliers options set to the default of "No special handling of outliers," and check the "Report each interpolated value option. Back to curve selection, You will find thata "Second Order Polynomial" is a good choice for ELISA standard curve data. But information about these and all other aspects of Prism, and data analysis in general, can be found in the Prism helpguide which can be accessed via button with a question mark. Clicking on that reveals the shape of the curve that will be fit using that selection, as well as some advanced options and settings, which you will not get into for the sake of time. If you click on one of the choices, the Details button appears. Notice to the right, there is a button called "Interpolate a standard curve." The dialog box that opens gives you a number of choices, and each one may be appropriate depending on the shape standard curve data. You can zoom in a little bit to see things more clearly. Also notice that the labels that we added to our data table are automatically added to our Graphas axis titles. You can see now that the Standard Curve data is here on your graph. You can select the plot with no connecting lines, because connecting lines would obscure the curve that you will fit to this data in the next step. Prism will ask us here to choose how you want your graph to look. Lets label this column "Concentration (ng/mL)", and this one "Absorbance (450 nm)."Some of you may be working with different units of concentration, or even log-concentrations,so simply label the columns accordingly. Now we will paste our standard curve data into the table.
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