An ATP Utilization Assay for Determining Bacterial Survival Under Antibiotics Treatment
Published: March 29, 2024
Abstract
Source: Wang, Liang-Chun, et al. Quantitative Examination of Antibiotic Susceptibility of Neisseria gonorrhoeae Aggregates Using ATP-utilization Commercial Assays and Live/Dead Staining. J. Vis. Exp. (2019).
This video describes the simple ATP-measuring assay using luciferase glow reagent. This assay is used to quantify Neisseria gonorrhoeae survival after treatment with ceftriaxone.
Protocol
1. Viability quantification of GC aggregations Collect GC using a sterile applicator. Swab GC from the plate and re-suspend GC in pre-warmed broth(GCP, Table 1) supplemented with 4.2% NaHCO3 and 1% Kellogg solutions. Use spectrophotometry at a wavelength of 650 nm (an OD 650 of 1 = ~1 x 109 CFU/mL) to determine the concentration of suspended bacteria. Adjust the concentration of GC to ~1 x 108 CFU/mL. Add 99 µL of adju…
Declarações
The authors have nothing to disclose.
Materials
| 100x Kellogg's supplement | |||
| Agar | United States Biological | A0930 | |
| BacTiter Assay | Promega | G8232 | |
| Ceftriaxone | TCI | C2226 | |
| Difco GC medium base | BD | 228950 | |
| Ferric nitrate, nonahydrate | Sigma-Aldrich | 254223-10G | |
| Glucose | Thermo Fisher Scientific | BP350-1 | |
| BacLight live/dead staining | Invitrogen | L7012 | |
| MS11 Neisseria gonorrhoeae strain | kindly provided by Dr. Herman Schneider, Walter Reed Army Institute for Research | ||
| Sonicator | Kontes | Equivelent to 9110001 |
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An ATP Utilization Assay for Determining Bacterial Survival Under Antibiotics Treatment. J. Vis. Exp. (Pending Publication), e22010, doi: (2024).