Table 2.
Effect of lidocaine injection on day 7 (peak of Borrelia multiplication) in the skin of C3H/HeN mice infected with B. afzelii NE4049.
| Mouse no. | Protocol | Number and mobility of Borrelia in BSK-H complete medium | PCR on Borrelia-infected mouse skin | ||
|---|---|---|---|---|---|
| Day 7 After biopsy | Day 14 After biopsy | Day 0 (Quantification flaB/gapdh gene) | Day 7 After biopsy (Ct) | ||
| 1 | Infected skin at day 7 | ++++ | ++++ | 173 | 20 |
| 2 | ++++ | ++++ | 212 | 17 | |
| 3 | ++++ | ++++ | 252 | 16 | |
| 4 | ++++ | ++++ | 189 | 20 | |
| 5 | ++++ | ++++ | 169 | 19 | |
| 6 | ++++ | ++++ | 187 | 16 | |
| 7 | Infected skin at day 7 + lidocaine | ++ | ++++ | 173 | 17 |
| 8 | ++ | ++++ | 182 | 19 | |
| 9 | ++ | ++++ | 191 | 19 | |
| 10 | +++ | ++++ | 197 | 19 | |
| 11 | +++ | ++++ | 237 | 19 | |
| 12 | +++ | ++++ | 271 | 16 | |
Mouse skin biopsy samples were collected at day 7 post-inoculation, and cultured for 7–14 days. To determine the spirochetes burden in skin biopsies, qPCR was also performed at day 7 without culture. After 7 days of culture, quantification of B. afzelii in the skin relative to host DNA was not possible due to damage to the skin in BSK-H medium, which reduced quality of mouse DNA, inhibiting gapdh amplification and detection. Therefore, Ct is provided for flaB only after 7 days of in vitro culture. Samples 3 and 12 marked in bold indicate two skin samples selected for proteomic analysis.