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J. Med. Microbiol. -- Vol. 51 (2002), 192-200
© 2002 Society for General Microbiology
ISSN 0022-2615

DIAGNOSTIC MICROBIOLOGY

Rapid antimicrobial susceptibility testing of urinary tract isolates and samples by flow cytometry

CHRISTIAN GAUTHIER*, YVES ST-PIERRE and RICHARD VILLEMUR

INRS-Institut Armand-Frappier, Laval (Québec), Canada H7V 1B7

Corresponding author: Dr R. Villemur (e-mail: richard. villemur{at}inrs-iaf.uquebec.ca).

Received 7 February 2001; revised version accepted 25 July 2001.

Abstract

A multiparametric flow cytometry antimicrobial susceptibility test was developed and its performance was evaluated on clinical urine isolates and samples in comparison with standard methods. Alterations in cytoplasmic membrane integrity were monitored by propidium iodide, and the anionic probe bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)) was used to measure changes in membrane potential. Microbial size and cellular content were analysed by light scattering. Twelve antibiotics were tested on 6 ATCC control strains, 22 urine isolates and 19 clinical urine samples, variously containing Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis, Staphylococcus aureus, S. saprophyticus and S. epidermidis. Agreement between the flow cytometry results, broth microdilution and disk diffusion tests was 93.9% (n=3285mutests). Of the 20 discrepancies observed, 18 were for species other than E. coli. Perfect correlation was obtained with five antibiotics, whereas norfloxacin, nitrofurantoin and tetracycline were responsible for 13(65%) of the 20 discrepancies.






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