Very rapid flow cytometric assessment of antimicrobial susceptibility during the apparent lag phase of microbial (re)growth.



Jindal, Srijan, Thampy, Harish, Day, Philip JR and Kell, Douglas B ORCID: 0000-0001-5838-7963
(2019) Very rapid flow cytometric assessment of antimicrobial susceptibility during the apparent lag phase of microbial (re)growth. Microbiology, 165 (4). 439 - 454.

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Abstract

Rapid changes in the number and flow cytometric behaviour of cells of E. coli taken from a stationary phase and inoculated into rich medium.Cells of E. coli were grown in LB medium, taken from a stationary phase of 2-4 h, and re-inoculated into fresh media at a concentration (105 ml-1 or lower) characteristic of bacteriuria. Flow cytometry was used to assess how quickly we could detect changes in cell size, number, membrane energization (using a carbocyanine dye) and DNA distribution. It transpired that while the lag phase observable macroscopically via bulk OD measurements could be as long as 4 h, the true lag phase could be less than 15-20 min, and was accompanied by many observable biochemical changes. Antibiotics to which the cells were sensitive affected these changes within 20 min of re-inoculation, providing the possibility of a very rapid antibiotic susceptibility test on a timescale compatible with a visit to a GP clinic. The strategy was applied successfully to genuine potential urinary tract infection (UTI) samples taken from a doctor's surgery. The methods developed could prove of considerable value in ensuring the correct prescription and thereby lowering the spread of antimicrobial resistance.

Item Type: Article
Uncontrolled Keywords: antimicrobial, susceptibility testing, flow cytometry, carbocyanine, proliferation
Depositing User: Symplectic Admin
Date Deposited: 14 Feb 2019 08:34
Last Modified: 30 Nov 2021 08:13
DOI: 10.1099/mic.0.000777
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3032799