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Antimicrobial Agents and Chemotherapy, September 2006, p. 2990-2995, Vol. 50, No. 9
0066-4804/06/$08.00+0     doi:10.1128/AAC.01511-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Detection of Extended-Spectrum ß-Lactamases in Clinical Isolates of Pseudomonas aeruginosa

Xiaofei Jiang, Zhe Zhang, Min Li, Danqiu Zhou, Feiyi Ruan, and Yuan Lu*

Center of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, People's Republic of China

Received 24 November 2005/ Returned for modification 25 January 2006/ Accepted 10 June 2006

With the occurrence of extended-spectrum ß-lactamases (ESBLs) in Pseudomonas aeruginosa being increasingly reported worldwide, there is a need for a reliable test to detect ESBLs in clinical isolates of P. aeruginosa. In our study, a total of 75 clinical isolates of P. aeruginosa were studied. Nitrocefin tests were performed to detect the ß-lactamase enzyme; isoelectric focusing electrophoresis, PCR, and PCR product sequencing were designed to further characterize the contained ESBLs. Various ESBL-screening methods were designed to compare the reliabilities of detecting ESBLs in clinical isolates of P. aeruginosa whose ß-lactamases were well characterized. Thirty-four of 36 multidrug-resistant P. aeruginosa clinical isolates were positive for ESBLs. blaVEB-3 was the most prevalent ESBL gene in P. aeruginosa in our study. Among the total of 34 isolates that were considered ESBL producers, 20 strains were positive using conventional combined disk tests and 10 strains were positive using a conventional double-disk synergy test (DDST) with amoxicillin-clavulanate, expanded-spectrum cephalosporins, aztreonam, and cefepime. Modifications of the combined disk test and DDST, which consisted of shorter distances between disks (20 mm instead of 30 mm) and the use of three different plates that contained cloxacillin (200 µg/ml) alone, Phe-Arg ß-naphthylamide dihydrochloride (MC-207,110; 20 µg/ml) alone, and both cloxacillin (200 µg/ml) and MC-207,110 (20 µg/ml) increased the sensitivity of the tests to 78.8%, 91.18%, 85.29%, and 97.06%.

* Corresponding author. Mailing address: Center of Laboratory Medicine, Huashan Hospital, Fudan University, 12 Central Urumqi Road, Shanghai 200040, People's Republic of China. Phone: 86-21-62498118. Fax: 86-21-62498118. E-mail: yuanlu{at}hsh.stn.sh.cn.

Antimicrobial Agents and Chemotherapy, September 2006, p. 2990-2995, Vol. 50, No. 9
0066-4804/06/$08.00+0     doi:10.1128/AAC.01511-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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