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Antimicrobial Agents and Chemotherapy, October 1998, p. 2474-2481, Vol. 42, No. 10
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Sequencing of Gyrase and Topoisomerase IV Quinolone-Resistance-Determining Regions of Chlamydia trachomatis and Characterization of Quinolone-Resistant Mutants Obtained In Vitro

Sophie Dessus-Babus, Cécile M. Bébéar, Alain Charron, Christiane Bébéar, and Bertille de Barbeyrac*

Laboratoire de Bactériologie, Université Bordeaux 2, 33076 Bordeaux Cedex, France

Received 5 February 1998/Returned for modification 11 May 1998/Accepted 6 June 1998

The L2 reference strain of Chlamydia trachomatis was exposed to subinhibitory concentrations of ofloxacin (0.5 µg/ml) and sparfloxacin (0.015 µg/ml) to select fluoroquinolone-resistant mutants. In this study, two resistant strains were isolated after four rounds of selection. The C. trachomatis mutants presented with high-level resistance to various fluoroquinolones, particularly to sparfloxacin, for which a 1,000-fold increase in the MICs for the mutant strains compared to the MIC for the susceptible strain was found. The MICs of unrelated antibiotics (doxycycline and erythromycin) for the mutant strains were identical to those for the reference strain. The gyrase (gyrA, gyrB) and topoisomerase IV (parC, parE) genes of the susceptible and resistant strains of C. trachomatis were partially sequenced. A point mutation was found in the gyrA quinolone-resistance-determining region (QRDR) of both resistant strains, leading to a Ser83right-arrowIle substitution (Escherichia coli numbering) in the corresponding protein. The gyrB, parC, and parE QRDRs of the resistant strains were identical to those of the reference strain. These results suggest that in C. trachomatis, DNA gyrase is the primary target of ofloxacin and sparfloxacin.

* Corresponding author. Mailing address: Laboratoire de Bactériologie, Université Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France. Phone: 33 5 56 79 56 67. Fax: 33 5 56 79 56 11. E-mail: Bertille.de.Barbeyrac{at}labbebear.u-bordeaux2.fr.

Antimicrobial Agents and Chemotherapy, October 1998, p. 2474-2481, Vol. 42, No. 10
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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