Guadalupe Miranda

Outbreak of Infection with Extended-Spectrum β-Lactamase-Producing Klebsiella pneumoniae in a Mexican Hospital

ABSTRACT 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 β-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 β-lactamase had an extended-spectrum β-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 β-lactamase.

Clonal and Horizontal Dissemination of Klebsiella pneumoniae Expressing SHV-5 Extended-Spectrum β-Lactamase in a Mexican Pediatric Hospital

ABSTRACT 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 β-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 β-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.

Antimicrobial Resistance from Enterococci in a Pediatric Hospital. Plasmids in Enterococcus faecalis Isolates with High-Level Gentamicin and Streptomycin Resistance

BACKGROUND Enterococcus spp. is an important nosocomial and community-acquired pathogen. Recent studies have documented the increasing importance of this pathogen in children, particularly in the hospital setting. Our objective in this study was to report the frequency of antimicrobial resistance in enterococci and to determine the characteristics of high-level gentamicin resistance (HLGR) plasmids in Enterococcus faecalis clinical isolates. METHODS Two hundred eighty-nine enterococcal isolates were collected during an 18-month period from a tertiary-care pediatric hospital in Mexico City. Isolates were screened for antibiotic resistance, including HLGR. High-level, gentamicin-resistant E. faecalis strains were selected for pulsed-field electrophoresis (PFGE) typing and plasmid analysis. Transferability of resistance markers was carried out using filter matings. RESULTS Seventy-six percent of isolates were E. faecalis, 10% were E. avium, 5.2% E. faecium, 5.2% E. raffinossus, 1.38% E. malodoratus, 0.6% E. hirae, and 0.6% E. casseliflavus. Antimicrobial resistance was ampicillin and penicillin 29%, imipenem 17%, and vancomycin 3%, HLGR 5%. The following 15 high-level, gentamicin-resistant isolates were identified: six E. faecalis; four E. avium; three E. faecium, and two E. casseliflavus. Five of the six E. faecalis isolates were different by PFGE and transferred gentamicin and streptomycin resistance on filter membranes. Transfer frequencies ranged from 8.2 x 10(-4) to 6.92 x 10(-5) transconjugants/recipient cell. The plasmid content of donors and transconjugants were homogeneous (one plasmid of 47 kb). CONCLUSIONS In this pediatric hospital, antimicrobial resistance in Enterococcus spp. is common. Frequency of high-level, gentamicin-resistant strains is low. Mechanism of HLGR appears to be due to a single plasmid dissemination.

Molecular Epidemiology of a Multiresistant Pseudomonas aeruginosa Outbreak in a Paediatric Intensive Care Unit

After isolation of multiresistant (MR) Pseudomonas aeruginosa from 3 hospitalized patients in a paediatric intensive care unit (PICU), a prospective surveillance programme was established to detect infected and/or colonized patients in the hospital. Isolates were examined by means of outer membrane protein (OMP) profiles, serotyping and DNA genomic analysis using pulsed-field gel electrophoresis (PFGE). Fifty-five P. aeruginosa strains were isolated from 23 hospitalized patients during September and October 1997. The median hospital stay before isolation of P. aeruginosa was 8 d. PFGE demonstrated that the same clone infected 14 patients, 4 of whom were not hospitalized in the PICU. Susceptibility patterns and OMP profiles correlated with PFGE results in 37.8% and 36.4% of cases, respectively. Serotype O11 correlated with pattern A in 77% of cases and serotype O4 correlated with unrelated strains in 75% of cases but did not discriminate between outbreak and unrelated isolates. Extensive investigation of cultures failed to identify a reservoir of P. aeruginosa. PFGE was superior to OMP analysis and serotyping for discriminating between strains. The possible mode of acquisition for most of the patients infected with the same clone was cross-contamination.

Response to Two Consecutive Protease Inhibitor Combination Therapy Regimens in a Cohort of HIV-1-infected Children

The response to 2 consecutive protease inhibitor (PI) combination regimens was evaluated in a cohort of HIV-1-infected children. Twelve children, most of whom had been heavily treated, received a 3-drug treatment: saquinavir in hard gelatin capsules (SQVhgc) + zidovudine (ZDV) + didanosine. When this treatment failed it was replaced by a 4-drug regimen: ritonavir + SQVhgc + ZDV + lamivudine. A mild and temporary decrease in viral load (VL) was observed with the initial regimen (p=0.22). Therapy failure occurred in 7 patients (58%) within 9 months and in another 3 (25%) within 9-18 months. The 7 children who failed within 9 months received the subsequent boosted regimen, leading to a significant and lasting reduction in VL ( p = 0.001). None of the patients failed on the boosted regimen: 5/7 achieved a VL of < 400 copies/ml and 3/7 achieved a VL of < 50 copies/ml. Our results suggest that a 4-drug regimen including 2 PIs produces a better and more sustained response than a 3-drug regimen including only 1 PI, and that a good, sustained response is possible with subsequent boosted regimens even in heavily treated children.

Fingerprint Analysis and Identification of Strains ST309 as a Potential High Risk Clone in a Pseudomonas aeruginosa Population Isolated from Children with Bacteremia in Mexico City

Pseudomonas aeruginosa is an opportunistic pathogen and is associated with nosocomial infections. Its ability to thrive in a broad range of environments is due to a large and diverse genome of which its accessory genome is part. The objective of this study was to characterize P. aeruginosa strains isolated from children who developed bacteremia, using pulse-field gel electrophoresis, and in terms of its genomic islands, virulence genes, multilocus sequence type, and antimicrobial susceptibility. Our results showed that P. aeruginosa strains presented the seven virulence genes: toxA, lasB, lecA, algR, plcH, phzA1, and toxR, a type IV pilin alleles (TFP) group I or II. Additionally, we detected a novel pilin and accessory gene, expanding the number of TFP alleles to group VI. All strains presented the PAPI-2 Island and the majority were exoU+ and exoS+ genotype. Ten percent of the strains were multi-drug resistant phenotype, 18% extensively drug-resistant, 68% moderately resistant and only 3% were susceptible to all the antimicrobial tested. The most prevalent acquired β-Lactamase was KPC. We identified a group of ST309 strains, as a potential high risk clone. Our finding also showed that the strains isolated from patients with bacteremia have important virulence factors involved in colonization and dissemination as: a TFP group I or II; the presence of the exoU gene within the PAPI-2 island and the presence of the exoS gene.