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. 2013 Jan 24;8(1):e54886. doi: 10.1371/journal.pone.0054886

Figure 2. Ratiometric pre-rRNA analysis of A. baumannii, S. aureus, and P. aeruginosa cells in serum.

Figure 2

A–C: Analysis of cells that had been held in serum for 7 days. Nutritional stimulation was initiated by suspending cells in pre-warmed TSB, and samples taken after 0, 1, 2, and 4 hours were subjected to RT-qPCR and qPCR to quantify pre-rRNA and gDNA, respectively. The same primers were used to amplify gDNA and cDNA generated from pre-rRNA. Ratios of pre-rRNA to gDNA (P:G; bars) are means and SDs of nine ratiometric permutations from three technical replicates of each sample type. Quantity of gDNA (lines) are means and standard deviations of the three gDNA measurements. Viable cell densities of A. baumannii, S. aureus, and P. aeruginosa, respectively, in serum were 9.0×108, 9.7×105, and <1×102 CFU/mL. From separate gDNA standard curves consisting of ≥ five points each, qPCR efficiencies were calculated [10(−1/slope) −1] to be between 0.913 and 0.959. A replicate experiment (Figure S1) yielded similar results for all three organisms.