Rapid Screening of Bacterial Antibiotic Resistance
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Corresponding Organization :
Other organizations : Liverpool School of Tropical Medicine, Aintree University Hospitals NHS Foundation Trust, University of Liverpool, Vector (United States), University of Salford, Dumfries and Galloway Royal Infirmary, NHS Dumfries and Galloway, Alder Hey Children's NHS Foundation Trust
Variable analysis
- Overnight cultures of the isolates
- Absorbance of the plate at OD450 every 39 seconds for 9 minutes and 45 seconds
- Optical density at 600 nm (OD600) of 0.1 in 10 ml of LB broth
- Centrifugation at 4,500 xg for 15 minutes
- Resuspension in 5ml of phosphate-buffered saline (PBS)
- Sonication on ice, for three intervals of 15 seconds, with a 30 second break between each sonication, using a Soniprep 150 plus (MSE centrifuges, UK)
- Addition of 90 μl of the supernatant to 10 μl of a 0.5 mg/ml nitrocefin solution in a 96 well clear, flat bottom plate
- Measurement using a FLUOstar OMEGA spectrophotometer (BMG lab systems, Germany)
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