We evaluated boronic acid (BA)-based methods for their ability to detect extended-spectrum ␤-lactamases (ESBLs) among clinical isolates of KPC-producing members of the Enterobacteriaceae family. A total of 155 isolates of Klebsiella pneumoniae (n ‫؍‬ 141), Escherichia coli (n ‫؍‬ 6), Enterobacter aerogenes (n ‫؍‬ 6), and Klebsiella oxytoca (n ‫؍‬ 2) genotypically confirmed to be KPC producers were analyzed. As many as 118 isolates harbored ESBLs (103 harbored SHV-type ESBLs, 13 harbored CTX-M-type ESBLs, and 2 harbored both SHV-and CTX-M-type ESBLs); the remaining 37 isolates were genotypically negative for ESBL production. The CLSI ESBL confirmatory test was positive for 79 of the 118 ESBL producers (sensitivity, 66.9%), while all 37 non-ESBL producers were negative (specificity, 100%). When a >5-mm increase in the zone diameter of either the cefotaxime (CTX)-clavulanate (CA) or the ceftazidime (CAZ)-CA disks containing BA compared with the zone diameter of the CTX or CAZ disks containing BA was considered to be a positive result for ESBL production, the method detected all 118 ESBL producers (sensitivity, 100%) and showed no false-positive results for non-ESBL producers (specificity, 100%). Double-disk synergy tests, in which disks of CTX, CAZ, aztreonam, or cefepime in combination with BA were placed at distances of 20, 25, and 30 mm (center to center) from a disk containing amoxicillin (amoxicilline)-clavulanate-BA, were able to detect 116 (98.3%), 101 (85.6%), and 28 (23.7%) of the ESBL-positive isolates, respectively; no false-positive results for non-ESBL-producing isolates were detected. Our results demonstrate that the modified CLSI ESBL confirmatory test with antibiotic disks containing BA is the most accurate phenotypic method for the detection of ESBLs in Enterobacteriaceae producing KPC carbapenemases.

a series of 45 outpatients presented with community-onset urinary tract infections due to carbapenem-resistant Pseudomonas aeruginosa isolates. Forty of them had a history of previous hospitalization or exposure to healthcare facilities, while the remaining five had not been previously admitted to our healthcare facilities or elsewhere within the preceding 12 months. In 18 outpatients, the carbapenem-resistant organisms caused recurrent community-onset urinary tract infections, while in three outpatients the organisms were also implicated in bacteremic episodes. All 45 single-patient P. aeruginosa isolates harbored the bla VIM-2 metallo-␤-lactamase (MBL) gene in a common class 1 integron structure. They belonged to one predominant pulsed-field gel electrophoresis type and three sporadically detected types; two of the sporadic clonal types were identified among outpatients without previous exposure to healthcare facilities, while the predominant clonal type was also identified to cause infections in hospitalized patients. This is the first study documenting that MBL-producing P. aeruginosa isolates cause community-onset infections that are related or not with exposure to healthcare facilities. Community-onset infections in our patients most likely resulted from the nosocomial acquisition of MBL producers, followed by a prolonged digestive carriage. The high rate of recurrent infections in the community underlies the difficulty of constraining infections caused by such microorganisms in the extrahospital setting.

The distribution of Chlamydia trachomatis serovars and Neisseria gonorrhoeae coinfection was studied in a group of 100 C. trachomatis-positive males with urethritis in Greece. The serovar distribution revealed that apart from the predominant worldwide types E and F, the relatively uncommon type G is also prevalent. Gonococcal coinfection was frequent (30%) and was associated with genovariant Ja (75%, P ‫؍‬ 0.008).Chlamydia trachomatis is the leading cause of bacterial sexually transmitted diseases (STDs) in industrialized countries, producing infections of the upper and lower genital tract in males and females, including urethritis, epididymitis, and proctitis in males (7,9,14,19). Currently, more than 18 different serovars of the organism have been identified based on conventional serotyping, while more than 29 variants have been recognized by employing monoclonal antibodies or genotypic methods (1, 15).The knowledge of the distribution profile of C. trachomatis urogenital serovars has been the focus of several studies in different regions worldwide, since it provides valuable information about their epidemiology and pathogenicity that contributes to the implementation of sufficient STD control measures. The most common serovar detected worldwide is E (up to 22 to 49% of cases) followed by serovars F and D (17 to 22% and 9 to 19%, respectively), while other serovars are less frequently identified (1,10,12,15,16,17).No data are available in Southern European countries, such as Greece, about the circulating C. trachomatis serovars among either general or specific populations, apart from a previous study in Italy that examined the C. trachomatis serovar distribution in a group of male patients with urethritis (4). The C. trachomatis serovars D through K, including the serovars Da and Ia and the genovariant Ja, are related to genital tract disease (2). Moreover, on a worldwide basis, although concomitant infection with Neisseria gonorrhoeae and C. trachomatis is well established and frequent according to epidemiological data (3,5,11,20), very little is known about the molecular biology-based identification of N. gonorrhoeae coexisting infection and its association with C. trachomatis serovars.The objectives of this study were to provide novel data regarding the C. trachomatis serovar distribution in this specific geographical area, by examining a specific group of symptomatic male patients with C. trachomatis urethritis living in Greece, and also to investigate the presence of N. gonorrhoeae coinfection by employing molecular methods and to discover any possible links of N. gonorrhoeae coinfection with particular C. trachomatis serovars.

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