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Antimicrobial Agents and Chemotherapy, February 2003, p. 636-642, Vol. 47, No. 2
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.2.636-642.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Methodologies and Cell Lines Used for Antimicrobial Susceptibility Testing of Chlamydia spp.

R. J. Suchland,* W. M. Geisler, and Walter E. Stamm

Division of Allergy and Infectious Diseases, School of Medicine, University of Washington, Seattle, Washington

Received 6 June 2002/ Returned for modification 11 October 2002/ Accepted 19 November 2002

In vitro susceptibility testing was performed on strains of Chlamydia trachomatis, Chlamydia pneumoniae, and Chlamydia psittaci under various conditions, including the cell line utilized, the time between infection and the addition of an antimicrobial, the concentration of inoculum, and the effect of multiple passage on the minimal chlamydicidal concentrations for the antibiotics doxycycline, azithromycin, erythromycin, ofloxacin, and tetracycline. With macrolides, the MIC varied depending upon the cell line utilized. With all antimicrobials, the MIC was related to the time at which the antimicrobial was added after infection. By an optimized cell culture passage method, all strains of chlamydia tested demonstrated survival after exposure to high levels (>100 times the MIC) of antimicrobials. Furthermore, upon retest, these surviving organisms did not demonstrate increased MICs. Thus, this phenomenon does not reflect selection of antimicrobial-resistant mutants but rather survival of some organisms in high antimicrobial concentrations (heterotypic survival). An additional 44 clinical isolates of C. trachomatis from patients with single-incident infections were tested against those from patients with recurrent or persistent infections, and heterotypic survival was seen in all isolates tested; hence, in vitro resistance did not correlate with the patient's apparent clinical outcome.

* Corresponding author. Mailing address: University of Washington School of Medicine, Division of Allergy and Infectious Diseases, Box 356523, 1959 NE Pacific St., Seattle, WA 98195. Phone: (206) 616-4170. Fax: (206) 616-4898. E-mail: wes{at}u.washington.edu.

Antimicrobial Agents and Chemotherapy, February 2003, p. 636-642, Vol. 47, No. 2
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.2.636-642.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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