Figure 1: Determination of the limit of detection for the Time Zero Test. GBS (a), Enterococcus (b), and N. gonorrhoeae (c) were prepared at 0.5 McFarland in PBS and diluted out serially. Following 20-minute incubation at room temperature, wells were washed and then HRP-conjugated antibody directed against the bacterium of interest was used to detect any bound organism. Three distinct strains of GBS were tested, two strains of Enterococcus, and N. gonorrhoeae each. Results shown are of GBS clinical isolate 01.12.76 (a), Enterococcus ATCC strain 29212 (b), and N. gonorrhoeae strain 19424 (c); each study was performed in triplicate and representative of three individual experiments. Bacterial concentrations were confirmed by CFUs following plating on agar for 24–48 hours.