Thirty-one strains of Klebsiella pneumoniae (including 10 duplicates) from 21 septicemic pediatric patients (age, <2 months) were studied during a 4-month period (June to October 1996) in which the fatality rate was 62% (13 of 21). These isolates identified by the API 20E system yielded the same biotype. Pulsed-field gel electrophoresis experiments revealed the same clone in 31 strains. The isolates were multidrug-resistant but were still susceptible to ciprofloxacin, imipenem, and cefoxitin. A 135-kb plasmid was harbored in all of the isolates. No transconjugants were obtained that were resistant to ampicillin, cefotaxime, tetracycline, or gentamicin. Isoelectric focusing for ␤-lactamases was performed on all strains, and three bands with pIs of 5.4, 7.6, and 8.2 were obtained. Of these, the pI 8.2 ␤-lactamase had an extended-spectrum ␤-lactamase phenotype. PCR amplification of both TEM-and SHV-type genes was obtained. The sequence analysis of the SHV PCR product indicated a mutation corresponding to the SHV-5 ␤-lactamase.Klebsiella pneumoniae is an important hospital-acquired pathogen with the potential of causing severe morbidity and mortality in pediatric patients. Several outbreaks of infection caused by K. pneumoniae isolates that are simultaneously resistant to broad-spectrum cephalosporins and aminoglycosides have been widely reported (8,9,18,21). Some of these multidrug-resistant isolates produce extended-spectrum ␤-lactamases (ESBLs) that are able to hydrolyze expanded-spectrum cephalosporins (e.g., ceftriaxone, cefotaxime, and ceftazidime), aztreonam, and related oxyimino-␤-lactams. Most of these enzymes are TEM-or SHV-type ␤-lactamases in which the substitution of one or more amino acids has altered the configuration of the active site (7,14,19). Most of the plasmids determining ESBLs are large (Ն80 kb) and encode multiple resistances (10). Little is known about the ESBL-producing Enterobacteriaceae in Mexico (23). The present work is a characterization of 31 ESBL-producing K. pneumoniae isolates from a nosocomial outbreak occurring in a hospital in Cuernavaca, Mexico. MATERIALS AND METHODSPatients and bacterial strains. The General Hospital in the state of Morelos is a secondary-care facility with 100 beds. The neonatal intensive care unit has 8 beds. An outbreak was suspected in the neonatal intensive care unit due to an increased number of isolates of K. pneumoniae from blood cultures during a 4-month period. A retrospective epidemiological investigation included 74 children less than 2 months old hospitalized from June to October 1996.Twenty-one clinical isolates were from blood; another 10 duplicate strains were from sites other than blood (urine and catheter tip) corresponding to 10 patients. In total there were 31 clinical isolates identified as K. pneumoniae by the API 20E system (BioMerieux, Merck).Susceptibility testing. The antimicrobial susceptibility was initially determined with the MicroScan (Dade) system, using the Combo 14 panel. Subsequently, MICs of several ␤-lactams were determine...

Serratia marcescens is a well-known cause of nosocomial infections and outbreaks, particularly in critically ill neonates and immunocompromised patients. Numerous methods have been proposed for typing. We used pulsed-field gel electrophoresis (PFGE) typing to analyze an outbreak in a neonatal intensive care unit (NICU). We included 23 patient isolates from an outbreak (March to July 1995), and 10 patient isolates from different wards during the same time period. PFGE of whole-cell DNA digested by SpeI was used as a marker of strain identity. The most common presentation of the infection was sepsis in 18 of 23 (78%) neonates. Only four different biotypes were identified; biotype A8d accounted for 84% of the strains. PFGE typing revealed two clones responsible for two different clonal strain dissemination outbreaks from March to July, with 24 patient isolates being pattern A and 4 patient isolates being pattern E. PFGE typing suggests cross transmission between patients in the NICU and other wards. The isolates from 5 other patients showed distinct PFGE patterns. Extensive investigation and cultures failed to identify any environmental or staff reservoir of S. marcescens. This is one of the first reports applying PFGE to the study of S. marcescens, and this method was a useful marker of strain identity. PFGE typing distinguished strains which appeared to be the same by biotyping.

One hundred eighty-four clinical isolates of Klebsiella pneumoniae were recovered from August 1996 to October 1997 at the Pediatric Hospital of the Instituto Mexicano del Seguro Social in Mexico City, Mexico. Most of the isolates were collected from the neonatal intensive care unit and infant wards, which are located on the same floor of the hospital. Isolates were genotypically compared by pulsed-field gel electrophoresis with XbaI restriction of chromosomal DNA. Of 184 clinical isolates, 91 belonged to cluster A and comprised three subtypes (A1, A2, and A3), while 93 isolates, comprising two minor clones, B (10 isolates) and C (7 isolates), and 76 unique patterns, were considered unrelated isolates (URI). Susceptibility patterns were indistinguishable in both groups. Fifty extended-spectrum ␤-lactamase-producing isolates, including 34 from clone A and 16 from URI, were examined for further studies. Molecular and genetic analysis showed that 47 of 50 clinical isolates expressed the SHV-5 ␤-lactamase. This enzyme, in combination with TEM-1, was encoded in a >170-kb conjugative plasmid. Results indicate that dissemination of this resistance was due to clonal and horizontal spread.