Assays were performed as outlined previously (20 (link), 31 (link), 32 (link)). Briefly, B. burgdorferi strain B314 producing B. hermsii FbpC (pAP2), as well as the vector-only control B314 pBBE22luc, were grown at 32°C in 1% CO2 in complete BSK-II media with kanamycin at 300 μg/ml to early-to mid-log phase. The cells were then diluted in complete BSK-II media to a final cell density of 1×106 cells/mL, then incubated for 1.5 hours in 15% normal human serum (Complement Technology) or equivalent heat inactivated serum as a positive control for survival. Cell survival was assessed by dark field microscopy based on cell motility and membrane damage or lysis.
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B. burgdorferi Serum Resistance Assay
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Corresponding Organization : Texas A&M University
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Variable analysis
independent variables
- Presence of B. hermsii FbpC (pAP2) in B. burgdorferi strain B314
- Cell survival in normal human serum
- Cell motility and membrane damage or lysis
- Growth conditions (32°C, 1% CO2, complete BSK-II media with kanamycin at 300 μg/ml)
- Initial cell density (1×10^6 cells/mL)
- Incubation time (1.5 hours)
- Positive control: Heat inactivated normal human serum
- Negative control: Vector-only control B314 pBBE22luc
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