Fig 5. Sensitivity and determination of the limits of detection (LOD) and quantitation (LOQ) for the eASTR4 test using synthetic template DNA.
A) Five-fold serial dilutions of a synthetic template DNA were added to the eASTR4 reaction at copy numbers ranging from 62,500 to 0.032 copies per reaction and the mean transformed Ct values (50.001-Ct) determined for each of the eight or 24 technical replicates. The whiskers depict the standard error of the mean. The NTC controls never had any amplification detected. The proportion of technical replicates that passed the Ct cut off for detection (% Positive) and number of technical replicates (n) measured are indicated at the top of the graph for each dilution tested. B) Percent binomial standard error determined at each dilution of synthetic template DNA for differing technical replicates: n = 3 (solid squares), n = 5 (solid circles), n = 8 (open circles), n = 12 (solid triangles), and n = 25 (open triangles).