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…

Açıklamalar

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

Etiketler

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Bu Makaleden Alıntı Yapın
An ATP Utilization Assay for Determining Bacterial Survival Under Antibiotics Treatment. J. Vis. Exp. (Pending Publication), e22010, doi: (2024).

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