Generation of bioluminescent <i>T. brucei</i> GVR35 for detection of trypanosomes <i>in vivo</i>.

<p>(<b>A</b>) Live GVR35 WT and -LUC2 clones were assessed for luciferase activity <i>in vitro</i> after addition of D-luciferin. The <i>in vitro</i> detection limit was determined by imaging a dilution series of GVR35-LUC2 using IVIS. Data show, on a logarithmic scale, average total flux (in photons per second) ± SD of replicate wells containing a specified trypanosome number. The dotted line indicates background bioluminescence for GVR35 WT. (<b>B</b>) Brains of mice infected with WT or GVR35-LUC2 were compared for trypanosome DNA and neuropathology at different times after infection. Real-time qPCR of the trypanosome TbPFR2 gene was performed on brain homogenates. Neuropathology was scored on haematoxylin and eosin stained sections. Graphs show means and 95% confidence interval for each group (<i>n</i> = 11–12 per group). <i>P</i> value (Student's <i>t</i> test) compared to GVR35 WT is indicated *<i>P</i><0.05; **<i>P</i><0.01.