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PubMed2005

Integron content of extended-spectrum-beta-lactamase-producing Escherichia coli strains over 12 years in a single hospital in Madrid, Spain.

The contribution of integrons to the dissemination of extended-spectrum beta-lactamases (ESBL) was analyzed on all ESBL-producing Escherichia coli isolates from 1988 to 2000 at Ramon y Cajal Hospital. We studied 133 E. coli pulsed-field gel electrophoresis types: (i) 52 ESBL-producing clinical strains (C-ESBL) (16 TEM, 9 SHV, 21 CTX-M-9, 1 CTX-M-14, and 5 CTX-M-10); (ii) 43 non-ESBL blood clinical strains (C-nESBL); and (iii) 38 non-ESBL fecal isolates from healthy volunteers (V-nESBL). Class 1 integrons were more common among C-ESBL (67%) than among C-nESBL (40%) or V-nESBL (26%) (P < 0.001) due to the high number of strains with bla(CTX-M-9), which is linked to an In6-like class 1 integron. Without this bias, class 1 integron occurrence would be similar in C-ESBL and C-nESBL groups (47% versus 40%). Occurrence of class 2 integrons was similar among clinical and community isolates (13 to 18%). No isolates contained class 3 integrons. The relatively low rate of class 1 integrons within transferable elements carrying bla(TEM) (23%) or bla(SHV) (33%) and the absence of class 2 integrons in all ESBL transconjugants mirror the assembly of translocative pieces containing bla(TEM) or bla(SHV) on local available transferable elements lacking integrons. The low diversity of class 1 integrons (seven types recovered in all groups) might indicate a wide dissemination of specific genetic elements in which they are located. In our environment, the spread of genetic elements encoding ESBL has no major impact on the dispersion of integrons, nor do integrons have a major impact on the spread of ESBL, except when bla(ESBL) genes are within an integron platform such as bla(CTX-M-9).

Cloning, Molecular↗
PubMed2003

Dio-Sensimedia: a novel culture medium for rapid detection of extended spectrum beta-lactamases.

BACKGROUND: Resistance to contemporary broad-spectrum beta-lactams, mediated by extended-spectrum beta-lactamases (ESBL), is an increasing problem worldwide. Many of the emerging antimicrobial resistance problems of this decade have been characterized by difficulty in the recognition of resistance in the laboratory, particularly by rapid susceptibility test methods. The plasmid-encoded ESBL represent such a resistance phenomenon that is difficult to recognize. We compared Dio-Sensimedia-ES (DSM-ES; Diomed, Istanbul, Turkey) and Mueller-Hinton (MH) agar in the double-disk synergy test (DDST) as a novel rapid system for detecting ESBL directly from bacterial culture. METHODS: Sixty ESBL-producing Klebsiella pneumoniae isolates cultured from blood (30), endotracheal aspirates (20), urine (5) and pus (5), as well as 40 Escherichia coli isolates cultured from endotracheal aspirates (15), urine (10), blood (8) and pus (7) were studied. Isolates positive for ESBL by the combined disk tests were tested with the DDST using MH and DSM-ES agar to detect ESBL-mediated resistance in K. pneumoniae and E. coli. DSM-ES agar was also used to determine the susceptibility of Enterobacteriaceae and staphylococci. RESULTS: Among 60 ESBL-producing K. pneumoniae isolates, 59 (98.3%) were identified as ESBL-positive by the DDST using MH, and 58 (96.6%), using DSM-ES agar. Of 40 ESBL-producing E. coli isolates, 38 (95%) were ESBL-positive by the DDST on MH agar, and 37 (92.5%), on DSM-ES agar. The average incubation period required for ESBL detection by the DDST on DSM-ES agar was 4 hours. CONCLUSIONS: Since the DDST results were available within 4 hours when DSM-ES agar was used, the use of this media may significantly lower the length of hospital stay, the total cost for patient care and even the mortality rate by facilitating early treatment against ESBL-producing organisms.

Agar↗
PubMed2006

Pseudomonas aeruginosa bloodstream infections: risk factors and treatment outcome related to expression of the PER-1 extended-spectrum beta-lactamase.

BACKGROUND: Bloodstream infection (BSI) due to Pseudomonas aeruginosa (Pa) has relevant clinical impact especially in relation to drug resistance determinants. The PER-1 extended-spectrum beta-lactamase (ESBL) is a common enzyme conferring high-level resistance to anti-pseudomonal cephalosporins. Risk factors and treatment outcome of BSI episodes caused by PER-1-positive Pa (PER-1-Pa) strains were compared to those caused by ESBL-negative Pa isolates (ESBL-N-Pa). METHODS: Twenty-six BSI cases due to ceftazidime-resistant Pa strains have been investigated. MIC values of anti-pseudomonal drugs were determined by the Etest method (AB Biodisk, Solna, Sweden). The double-disk synergy test was used to detect ESBL production. PCR amplification and DNA sequencing were used to characterize ESBL types. Clinical records of BSI-patients were examined retrospectively. Demographic data, underlying diseases (McCabe-Jackson classification and Charlson weighted index), risk factors, antimicrobial therapy, and treatment outcome were evaluated in cases due to ESBL-positive and cases due to ESBL-N-Pa isolates. Unpaired Student's t-test, Mann-Whitney U-test, Fisher's exact test and the chi2 test were used for statistical analysis. RESULTS: Nine Pa isolates expressed the PER-1 ESBL; the remaining 17 isolates did not produce ESBLs. Severe sepsis (P = 0.03), bladder and intravascular catheters (both P = 0.01), immunosuppressive therapy (P = 0.04), and mechanical ventilation (P = 0.03) were significantly associated with BSI due to PER-1-Pa. Empirical treatment (P = 0.02) and treatment after ID/AST (P < 0.01) were rarely adequate in PER-1-Pa cases. With regard to treatment outcome, 77.8% BSI cases due to PER-1-Pa vs. 28.6% cases due to ESBL-N-Pa isolates failed to respond (P < 0.03). All cases due to PER-1-Pa that were treated with carbapenems (alone or in combination with amikacin) failed to respond. In contrast, 7/8 cases due to ESBL-N-Pa given carbapenems were responders. CONCLUSION: Therapeutic failure and increased hospital costs are associated with BSI episodes caused by PER-1-Pa strains. Thus, recognition and prompt reporting of ESBL-production appears a critical factor for the management of patients with serious P. aeruginosa infections.

Adult↗
PubMed2006

Occurrence of a novel SHV-type enzyme (SHV-55) among isolates of Klebsiella pneumoniae from Portuguese origin in a comparison study for extended-spectrum beta-lactamase-producing evaluation.

Fifty-five isolates of Klebsiella pneumoniae were evaluated for extended-spectrum beta-lactamase (ESBL) detection and confirmation, using MIC testing by agar dilution, broth microdilution, and the ESBL E-Test (AB Biodisk, Solna, Sweden), according to reference laboratory criteria (RLC) and Clinical and Laboratory Standards Institute (CLSI) guidelines. The RLC classify as ESBL producers those strains for which any MIC of cephalosporins is 3-fold lower in the presence of 2 mug/mL of clavulanate. The E-Test was the only to show 100% sensitivity and specificity to detect ESBL-producer strains with either set of guidelines. MIC determination by agar dilution or broth microdilution, using NCCLS guidelines, showed sensitivity of 92.9%. Nucleotide sequencing allowed the identification of a new ESBL (SHV-55). Overall, this gold standard method confirmed the production of 18 ESBL producers, 36 non-ESBL producers, from which 9 were false ESBL producers (suggesting hyperproduction) and 1 presumptive ESBL TEM-derived. New guidelines for ESBL detection and reliable methods of ESBL identification are required.

Base Sequence↗
PubMed2002

Investigation of extended-spectrum beta-lactamase in Klebsiellae pneumoniae and Escherichia coli from China.

Extended-spectrum beta-lactamases (ESBLs) are an increasing cause of resistance to third-generation cephalosporins in Enterobacteriaceae. However, they have not been well studied in China. We investigated the prevalence, resistance, and probable gene type of ESBLs using MICs testing and polymerase chain reaction in 559 Klebsiellae pneumoniae and 427 Escherichia coli isolates collected from patients in Huashan Hospital from 1 January to 31 December 1999. The incidence of ESBL-producing strains was 51% among Klebsiellae pneumoniae (285/559) and 23.6% among Escherichia coli (101/427), most of which were collected from patients in intensive care unit and neurosurgical ward. PFGE showed that some epidemic ESBL-producing strains were present in the ICU, especially among ESBL-producing Klebsiellae pneumoniae. The major source of ESBL-producing Klebsiellae pneumoniae and Escherichia coli was sputum specimen (63.5%) and urine (64.3%), respectively. These strains were resistant to most beta-lactams (including the third-generation cephalosporins and monobactams) and non-beta-lactams (such as fluoroquinolones, aminoglycosides, tetracycline, and chloramphenicol), all or most ESBL producers were susceptible to imipenem, cefmetazole and beta-lactam/clavulanic acid. TEM was the main type of beta-lactamases and the CTX-M type of ESBLs was common in these isolates. Some ESBL-producing Escherichia coli and most ESBL-producing Klebsiellae pneumoniae produced more than one type of beta-lactamase. These data confirm that ESBL producers are common among hospital strains of Escherichia coli and Klebsiellae pneumoniae in China. It is important to monitor such strains closely and prevent their spread.

Anti-Bacterial Agents↗
PubMed2005

Clinical features of nosocomial infections by extended-spectrum beta-lactamase-producing Enterobacteriaceae in neonatal intensive care units.

AIM: To determine the risk factors for the acquisition of nosocomial extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae infection in infants hospitalized in neonatal intensive care units (NICUs) and to evaluate the therapeutic outcome of these infants. METHODS: We retrospectively reviewed the medical records of infants with nosocomial ESBL-producing Enterobacteriaceae infection hospitalized in NICUs at Chang Gung Children's Hospital in 2001. The clinical features of these infants were compared with a cohort of non-ESBL-producing Enterobacteriaceae-infected infants during the same period. The therapeutic outcome of the infants in the ESBL group was analysed. RESULTS: Seventy infants were included in this study. Thirty-one infants with 34 isolates were identified in the ESBL group and 39 infants with 42 isolates in the non-ESBL group. Of the parameters analysed, including gestational age, birthweight, length of hospital stay before onset, the number of antibiotics used, the duration of third-generation cephalosporin usage and the number of patients receiving a third-generation cephalosporin prior to the onset of infection, no significant difference was found between the two groups. The infection-contributed case fatality rate was 3.0% (1 of 33) in the ESBL group, not significantly different from that in the non-ESBL group (1 of 41, 2.4%). Of the 31 patients in the ESBL group, 18 were treated with a carbapenem as definitive therapy while 13 were treated with a non-carbapenem antibiotic regimen. No significant difference was noted in terms of mortality rate between the two subgroups. CONCLUSION: The outcome of the infants hospitalized in the NICU with ESBL-producing enterobacterial infections was not indispensably grave, even when treated with a non-carbapenem antibiotic regimen. The risk factors for the acquisition of ESBL-producing enterobacterial infections in these infants were not identified in this series.

Anti-Bacterial Agents↗
PubMed2004

Co-carriage rates of vancomycin-resistant Enterococcus and extended-spectrum beta-lactamase-producing bacteria among a cohort of intensive care unit patients: implications for an active surveillance program.

OBJECTIVE: To assess the co-colonization rates of extended-spectrum beta-lactamase (ESBL)-producing bacteria and vancomycin-resistant Enterococcus (VRE) obtained on active surveillance cultures. DESIGN: Prospective cohort study. SETTING: Medical and surgical intensive care units (ICUs) of a tertiary-care hospital. PATIENTS: Patients admitted between September 2001 and November 2002 to the medical and surgical ICUs at the University of Maryland Medical System had active surveillance perirectal cultures performed. Samples were concurrently processed for VRE and ESBL-producing bacteria. RESULTS: Of 1,362 patients who had active surveillance cultures on admission, 136 (10%) were colonized with VRE. Among these, 15 (positive predictive value, 11%) were co-colonized with ESBL. Among the 1,226 who were VRE negative, 1,209 were also ESBL negative (negative predictive value, 99%). Among the 1,362 who had active surveillance cultures on admission, 32 (2%) were colonized with ESBL. Among these, 15 (47%) were co-colonized with VRE. Of the 32 patients colonized with ESBL, 10 (31%) had positive clinical cultures for ESBL on the same hospital admission. For these 10 patients, the surveillance cultures were positive an average of 2.7 days earlier than the clinical cultures. CONCLUSIONS: Patients who are colonized with VRE can also be co-colonized with other antibiotic-resistant bacteria such as ESBL-producing bacteria. Our study is the first to measure co-colonization rates of VRE and ESBL-producing bacteria. Isolating VRE-colonized patients would isolate 47% of the ESBL-colonized patients without the need for further testing. Hence, active surveillance for VRE should also theoretically diminish the amount of patient-to-patient transmission of ESBL-producing bacteria.

Carrier State↗
PubMed2006

Extended-spectrum beta-lactamase genes of Klebsiella pneumoniae strains in Taiwan: recharacterization of shv-27, shv-41, and tem-116.

Klebsiella pneumoniae causing primary liver abscess (PLA) is emerging. This study identified the beta-lactamases genes of K. pneumoniae isolates in Taiwan. The susceptibilities of beta-lactam antibiotics of 30 K. pneumoniae strains associated with primary liver abscess and 30 noninvasive strains were analyzed. The beta-lactamase genes of randomly selected 24 strains from community-acquired infection and 7 extended-spectrum beta-lactamases (ESBL) strains were identified by PCR and DNA sequencing. Protein expression and the ESBL phenotype of beta-lactamase were determined. All 60 strains were ampicillin resistant and cefotaxime susceptible, whereas no strain was ESBL producing. In the 24 selected strains, shv-1a was found in 14, shv-1 in 7; shv-26, shv-27, and shv-41 were detected in one. However, all of these 24 strains had the tem-116 gene. In 7 ESBL-producing K. pneumoniae strains, shv-5a was found in 5, whereas shv-5 and ctx-m-9 group were detected in 1 strain. Two previously reported ESBL genes, shv-27 and tem-116, as well as a suspected ESBL gene, shv-41, were found in non-ESBL-producing strains. Transformation of these genes conferred ampicillin resistance but not the ESBL-producing phenotype in Escherichia coli. Beta-lactamase protein expression of these strains was further confirmed by western blotting. In conclusion, ESBL is rare in community-acquired K. pneumoniae infection and is not associated with PLA in Taiwan. The shv-5a, shv-5, and ctx-m-9 groups are present in ESBL-producing strains in Taiwan, but shv-27, shv-41, and tem-116 are not ESBL genes.

Cloning, Molecular↗
PubMed2005

Risk factors associated with extended-spectrum beta-lactamase-producing organisms at a tertiary care hospital.

BACKGROUND: In 1995, beta-lactam inhibitor combinations replaced third-generation cephalosporins as empirical therapy in an effort to manage extended-spectrum beta-lactamase (ESBL) resistance. This study investigated the relationship between antibiotic usage and ESBL organisms from 1994 through 2002 using epidemiological and molecular analysis. METHODS: A case-control study of 119 patients with ESBL organisms and 132 patients with non-ESBL organisms was conducted. Demographics, co-morbidities, device utilization and antibiotic use were analysed for all patients and infected patients only (cases = 75, controls = 83). Both exposure and degree of exposure (in grams) to antibiotics were included. A dot blot hybridization technique was used to identify genes in plasmid extracts from the ESBL organisms. RESULTS: Ventilator days OR 1.1 (1.06, 1.15) P < 0.001, adult respiratory distress syndrome (ARDS) OR 3.1 (1.0, 9.7) P = 0.05, prior aminoglycoside use OR 2.7 (1.2, 6.1) P = 0.02, prior third-generation cephalosporin use OR 7.2 (2.6, 20) P < 0.001, and prior trimethoprim/sulfamethoxazole use OR 8.8 (3.1, 26) P < 0.001 were significantly associated with ESBL organisms by multivariate analysis. All models were concordant with a significant association of ventilator days, third-generation cephalosporins and trimethoprim/sulfamethoxazole with ESBL organisms. beta-Lactamase inhibitor combinations were not associated with ESBL organisms. Hybridization of plasmid extracts demonstrated that 95% of the ESBL organisms carried intI1, a mobile DNA element with a sulphonamide-resistance (R) gene and a frequent carrier of other R factors. Genes for specific types of trimethoprim-R and aminoglycoside-R were present in 26% and 40% of the extracts, respectively. CONCLUSIONS: These data indicate that, besides patient risk factors and third-generation cephalosporins, other antibiotics may provide selective pressures in maintaining ESBL organisms due to multiple resistance genes on plasmids. beta-Lactamase inhibitor combinations appear to be an acceptable substitute to third-generation cephalosporins in strategies to control ESBL organisms.

Anti-Bacterial Agents↗
PubMed2002

Identification and minisequencing-based discrimination of SHV beta-lactamases in nosocomial infection-associated Klebsiella pneumoniae in Brisbane, Australia.

Extended-spectrum beta-lactamases (ESBLs) are active against oxyimino cephalosporins and monobactams. Twenty-one Klebsiella pneumoniae isolates obtained between 1991 and 1995 at the Princess Alexandra Hospital in Brisbane, Australia, were subject to amplification and sequencing of the SHV beta-lactamase-encoding genes. Thirteen strains were phenotypically ESBL positive. Of these, six strains carried the blaSHV-2a gene and seven strains carried the blaSHV-12 gene. Eight strains were phenotypically ESBL negative. Of these, seven strains carried the non-ESBL blaSHV-11 gene and one strain carried the non-ESBL blaSHV-1 gene. There was complete correspondence between the ESBL phenotype and the presence or absence of an ESBL-encoding gene(s). In addition, it was determined that of the 13 ESBL-positive strains, at least 4 carried copies of a non-ESBL-encoding gene in addition to the blaSHV-2a or blaSHV12 gene. A minisequencing-based assay was developed to discriminate the different SHV classes. This technique, termed "first-nucleotide change," involves the identification of the base added to a primer in a single-nucleotide extension reaction. The assay targeted polymorphisms at the first bases of codons 238 and 240 and reliably discriminated ESBL-positive strains from ESBL-negative strains and also distinguished strains carrying blaSHV-2a from strains carrying blaSHV-12. In addition, this method was used to demonstrate an association between the relative copy numbers of blaSHV genes in individual strains and the levels of antibiotic resistance.

Australia↗
PubMed2003

Characterization of clinical isolates of Enterobacteriaceae from Italy by the BD Phoenix extended-spectrum beta-lactamase detection method.

Production of extended-spectrum beta-lactamases (ESBLs) is an important mechanism of beta-lactam resistance in Enterobacteriaceae: Identification of ESBLs based on phenotypic tests is the strategy most commonly used in clinical microbiology laboratories. The Phoenix ESBL test (BD Diagnostic Systems, Sparks, Md.) is a recently developed automated system for detection of ESBL-producing gram-negative bacteria. An algorithm based on phenotypic responses to a panel of cephalosporins (ceftazidime plus clavulanic acid, ceftazidime, cefotaxime plus clavulanic acid, cefpodoxime, and ceftriaxone plus clavulanic acid) was used to test 510 clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Providencia stuartii, Morganella morganii, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Citrobacter freundii, and Citrobacter koseri. Of these isolates, 319 were identified as ESBL producers, and the remaining 191 were identified as non-ESBL producers based on the results of current phenotypic tests. Combined use of isoelectric focusing, PCR, and/or DNA sequencing demonstrated that 288 isolates possessed bla(TEM-1)- and/or bla(SHV-1)-derived genes, and 28 had a bla(CTX-M) gene. Among the 191 non-ESBL-producing isolates, 77 isolates produced an AmpC-type enzyme, 110 isolates possessed TEM-1, TEM-2, or SHV-1 beta-lactamases, and the remaining four isolates (all K. oxytoca strains) hyperproduced K1 chromosomal beta-lactamase. The Phoenix ESBL test system gave positive results for all the 319 ESBL-producing isolates and also for two of the four K1-hyperproducing isolates of K. oxytoca. Compared with the phenotypic tests and molecular analyses, the Phoenix system displayed 100% sensitivity and 98.9% specificity. These findings suggest that the Phoenix ESBL test can be a rapid and reliable method for laboratory detection of ESBL resistance in gram-negative bacteria.

Algorithms↗
PubMed2005

Clinical and molecular analysis of extended-spectrum {beta}-lactamase-producing enterobacteria in the community setting.

During a previous survey, five extended-spectrum beta-lactamase (ESBL)-producing enterobacteria (ESBLE) (two Enterobacter aerogenes isolates expressing TEM-24 b, two Escherichia coli isolates expressing TEM-21 or TEM-24 b, and one Klebsiella pneumoniae isolate expressing SHV-4/TEM-15) responsible for urinary tract infections (UTIs) were found among 1,584 strains collected from community patients. The aim of the present study was to elucidate the route of emergence of these typically nosocomial organisms in the community. Thus, the files of the five patients were analyzed over at least a decade, and potentially related ESBLE from hospitals or clinics were examined. Their enzymes were characterized at a molecular level, and the strains were typed by amplified-primed PCR, enterobacterial repetitive intergenic consensus PCR, and restriction plasmid profile. All patients (C1 to C5) had risk factors for ESBLE acquisition, including past history of hospitalization (2.5 to 23 months before). Four (C1 and C3 to C5) had previously received antibiotics (concomitantly to 35 months earlier), two (C1 and C3) had indwelling urinary catheters and recurrent UTIs, and three (C2, C3, and C5) formerly experienced ESBLE-induced UTIs (2 to 11 months before). The same ESBLE and/or an identical or similar ESBL-encoding plasmid was identified in the hospital ward (C1 to C4) or in a clinic (C5) where the patients had previously resided. Patients C1 and C4, infected with different ESBLE carrying a closely related plasmid, were hospitalized in the same unit. Persistence of ESBLE over at least a 5-year period was demonstrated for patient C3. Thus, community-acquired UTIs in these patients likely resulted from nosocomially acquired ESBLE or an ESBL-encoding plasmid followed by a prolonged digestive carriage.

Aged↗
PubMed2006

Extended spectrum beta-lactamase producing Enterobacteriaceae in Lebanese ICU patients: epidemiology and patterns of resistance.

Our study aims to investigate and describe the epidemiology of the intestinal carriage of ESBL-PS in intensive care units of five Lebanese hospitals and to analyze the potential risk factors for the acquisition of these strains. At the same time, we intend to determine the patterns of susceptibility of these strains, exploring therefore the availability of alternative treatment. One thousand, four hundred forty-two fecal samples were collected between January 1, 2003 and March 31, 2003 from 378 patients admitted to the ICUs of five Lebanese tertiary care general hospitals located in different areas of Lebanon. ESBL production was detected by the double disk synergy test and antibiotic susceptibility of ESBL-producing strains as well as minimum inhibitory concentrations were determined. A paired case-control study was undertaken to identify risk factors for carriage of ESBL-PS. One hundred eighteen strains isolated from 72 subjects were identified as ESBL producers, including 95 (80.5%) E. coli, 16 (13.6%) Klebsiella pneumoniae, and 7 (5.6%) Enterobacter cloacae. A higher rate of multiple ESBL-PS carriage was described among these acquisition cases (21 double carriages and 3 triple carriages of ESBL-PS compared to only 1 double carriage of ESBL-PS at admission). In general, similar trends of susceptibility were observed in the different hospitals. As expected, the lowest MIC was observed with imipenem for all E. coli, Klebsiella, and Enterobacter isolates. Ciprofloxacin, followed by trimethoprim-sulfamethoxazole seem to be associated with the lowest susceptibility. In vitro susceptibility to cefoxitin for all isolates was 74.6%; more resistance was associated to ceftazidime (90.7%) than to cefotaxime (69.7%). Our data agree with other national and international reports showing the increase in ESBL-PS carriage in ICU patients. They demonstrate the endemic character of this carriage in Lebanese hospitals and the important risk factors including immunosuppression and evidence of ESBL infection. The highly resistant profile of ESBL-PS to antimicrobial agents available for treatment reflects the severity of this issue. The significance of this study resides in the direct correlation between our results and the nationwide increase in multi-drug resistant bacteria and the continuous change in bacterial resistance epidemiology. Our data may have an important impact on infection control policies in hospitals and on treatment of infectious diseases.

Case-Control Studies↗
PubMed2002

[Extended-spectrum beta-lactamase in Klebsiella pneumoniae and Escherichia coli isolates].

OBJECTIVE: To investigate the prevalence, drug resistance, gene typing, and epidemicity of extended-spectrum beta-lactamases (ESBL) Klebsiella pneumoniae and Escherichia coli isolates. METHODS: 559 strains of K. pneumoniae and 427 strains of E.coli were isolated form Huanshan Hospital from 1 January to 31 December 1999. The ESBL-producing strains were detected by double disc test and confirmed by minimal inhibitory concentration (MIC). The MIC of ESBL-producing strains was detected by agar dilution test. The beta-lactamase genes were detected by PCR. DNA fingerprinting was made by pulsed-field gel electrophoresis (PFGE). RESULTS: The incidence of ESBL-producing strains was 51% among the isolated K. pneumoniae (285/559) and 23.6% among the isolated E. coli (101/427), most of which were collected from the patients in the intensive care unit and neurosurgical ward. 63.5% of the ESBL-producing K. pneumoniae strains were collected from sputum specimens, and 64.3% of the ESBL-producing E. coli strains were collected from the urine specimens. Most ESBL-producing strains were resistant to most beta-lactam antibiotics, including the third-generation cephalosporins, and non- beta-lactam antimicrobial drugs, such as fluoroquinolones, aminoglycosides, tetracycline, and chloramphenicol. Most of the ESBL-producing strains were susceptible to imipenam, cefmetazole, and beta-lactam antibiotic/clavulanic acid. TEM type beta-lactamase was the main type among those EBSL-producing strains, followed by SHV type and CTX-M type. Some ESBL-producing E. coli and most ESBL-producing K. pneumoniae produced more than one type of beta-lactamase. CONCLUSION: ESBL-producing strains are common among hospital strains of E. coli and K. pneumoniae. Most of them are multidrug resistant. Prevalence and transmission of these strains exist in hospital.

Anti-Bacterial Agents↗
PubMed2004

Prevalence of extended spectrum beta-lactamase among multidrug resistant gram-negative isolates from a general hospital in Saudi Arabia.

OBJECTIVE: To determine the prevalence of extended spectrum beta-lactamase (ESBL) among multidrug resistant isolates of enterobacteriaceae and non-fermenting gram-negative bacilli. METHODS: This study was carried out at the Almana General Hospital, Eastern Province, Kingdom of Saudi Arabia, during the period March 2002 through to June 2003. Multidrug resistant gram-negative isolates from patients admitted to the surgical, medical, pediatric, long-term care and intensive care units were studied for the presence of the ESBL enzyme. RESULTS: A total of 3231 gram-negative organisms were studied for the presence of multidrug resistance and ESBLs. Of these, 197 (6%) isolates were multidrug resistant (MDR), and 156 (4.8%) were positive for ESBL. Seventy nine percent of the MDR strains were positive for ESBL. The most frequent isolates were Escherichia coli (1116) and Klebsiella pneumoniae (687) and ESBL was detected in 72 (6.5%) and 37 (5.4%) of these isolates. The MDR strains that produced ESBL were most commonly isolated from surgical care patients with diabetic fascitis (83%) and patients with indwelling Foley's catheter (79%). Extended spectrum beta-lactamase producing strains showed the highest susceptibility to imipenem and meropenem (86%). The non-beta-lactam antibiotics with greatest activity against these ESBL strains in vitro were ciprofloxacin (72%), amikacin (70%), tobramycin (67%) and gentamicin (56%). CONCLUSION: The majority (79%) of the MDR enterobacteriaceae and non-fermenting gram-negative bacilli tested over 15-months were positive for ESBL. Imipenem, meropenem, ciprofloxacin and amikacin showed the highest activity against these ESBL-producing organisms. Due to the growing problem of infection with ESBL-producing bacteria, which are frequently resistant to many classes of antibiotics resulting in difficult-to-treat infections, clinicians need to be familiar with the clinical significance of these enzymes and potential strategies for dealing with them.

Adolescent↗
PubMed2006

Extended-spectrum beta-lactamases in Taiwan: epidemiology, detection, treatment and infection control.

Extended-spectrum beta-lactamases (ESBLs) efficiently hydrolyze extended-spectrum beta-lactams such as cefotaxime, ceftriaxone, ceftazidime, and aztreonam. ESBLs are most often plasmid-mediated. In Taiwan, the prevalence of ESBLs in bacteria has risen, ranging from 8.5 to 29.8% in Klebsiella pneumoniae and 1.5 to 16.7% in Escherichia coli isolates. The most prevalent types of ESBLs are SHV-5, SHV-12, CTX-M-3, and CTX-M-14 in isolates of K. pneumoniae and E. coli, with differences between institutions. SHV-12 and CTX-M-3 have been reported as the most common ESBLs in isolates of Enterobacter cloacae and Serratia marcescens, respectively. Molecular epidemiology studies suggest that the ESBL-encoding genes have been disseminated either by proliferation of epidemic strains or by transfer of plasmids carrying the resistance traits. The current ESBL screen guidelines of the Clinical and Laboratory Standards Institute (formerly National Committee for Clinical Laboratory Standards) are issued for E. coli, Klebsiella spp., and Proteus mirabilis. Owing to the lack of standard methods, it remains difficult to assure the presence of ESBL in an isolate co-harboring an AmpC beta-lactamase, particularly in cases where the latter is produced in larger amounts than the former. Empirical therapy with piperacillin-tazobactam to replace third-generation cephalosporins may help to reduce the occurrence of ESBLs in an institution with a high prevalence of ESBL producers. Carbapenems remain the drugs of choice for serious infections caused by ESBL-producing organisms. To retard the selection for carbapenem-resistant bacteria, 7-alpha-methoxy beta-lactams or fourth-generation cephalosporins can be therapeutic alternatives for mild-to-moderate infections provided that the pharmacokinetic and pharmacodynamic target can be easily achieved.

Carbapenems↗
PubMed1998

Antimicrobial susceptibility testing for Klebsiella pneumoniae isolates resistant to extended-spectrum beta-lactam antibiotics.

Detection of Klebsiella pneumoniae strains with extended-spectrum beta-lactamase (ESBL)-related resistance phenotypes is becoming important in clinical microbiology laboratories. In this study, we investigated the usefulness of three screening methods, the Etest ESBL screen, the double-disk synergy test, and the ceftazidime disk test, for identifying ESBL-producing K. pneumoniae strains. The agar dilution method was used as the standard. We also determined the in vitro activity of several new antimicrobial agents against these organisms. Strains that exhibited an increase in the minimum inhibitory concentration (MIC) to the third-generation cephalosporins or aztreonam of 2 micrograms/mL or more, but were susceptible to the three cephamycins tested, were considered to have ESBL-related resistance phenotypes. The frequency of ESBL-producing K. pneumoniae isolates (according to the disk-diffusion method) has increased markedly in recent years, from 3.4% in 1993 to 10.3% in 1997. A total of 93 preserved isolates of K. pneumoniae collected from December 1995 through March 1997 were found to be resistant to at least one of the third-generation cephalosporins (cefotaxime and ceftazidime) or aztreonam using the routine disk diffusion method. Among these isolates, 35 were classified as having an ESBL phenotype using the agar dilution method. The remaining 58 isolates were classified as cephamycin resistant, which indicated resistance to both cephamycins and third-generation cephalosporins or aztreonam. The susceptibility rates of the ESBL-producing isolates were 11% for cefotaxime, 14% for ceftazidime, and 6% for aztreonam. The susceptibility rates of these 35 isolates to imipenem, ciprofloxacin, and ofloxacin were 100%, 80%, and 86%, respectively. Both the MIC50 and MIC90 of meropenem were 0.06 microgram/mL, while the MIC50 and MIC90 of BAY 12-8039 were 0.125 and 2 micrograms/mL, respectively. Thirty-two (91%) of the 35 isolates of K. pneumoniae with the ESBL-related resistance phenotype were detected by the Etest ESBL screen, while the ceftazidime disk screen test detected 77% of these isolates, and the double-disk synergy test detected 74%. The Etest ESBL screen appears to be an acceptable, convenient, and sensitive method for the detection of ESBL-producing isolates in the clinical microbiology laboratory.

Anti-Infective Agents↗
PubMed2006

Resistance of strains producing extended-spectrum beta-lactamases and genotype distribution in China.

OBJECTIVE: To investigate the resistance of Escherichia coli and Klebsiella pneumoniae producing extended-spectrum beta-lactamases (ESBLs) and the genotyping of ESBLs in China. METHODS: MICs of 12 antibiotics against 50 strains (by random selection) of ESBLs-producing E. coli and K. pneumoniae were determined by E-test. The genotypes of ESBLs were analyzed by PCR, DNA sequencing and isoelectric focusing. RESULTS: The susceptibility rate of 50 isolates was 100% in imipenem, 60%-80% in cefoperazone/sulbactam, ceftazidime and piperacillin/tazobactam, and lower in other antimicrobial agents tested. Only 6.0% of the isolates were sensitive to cefotaxime. Four hundred and forty-seven of 509 isolates had been confirmed the genotype of ESBLs. Four hundred and sixteen strains produced only one type of ESBLs, including CTX-M-14 (271 strains), CTX-M-3 (70 strains), CTX-M-24 (35 strains), CTX-M-22 (8 strains), CTX-M-15 (4 strains), CTX-M-9 (4 strains), CTX-M-28 (3 strains), CTX-M-12 (1 strain), CTX-M-13 (1 strain), CTX-M-27 (1 strain), CTX-M-29 (1 strain), SHV-12 (10 strains), SHV-5 (4 strains), SHV-2 (2 strains), and SHV-9 (1 strain). Thirty isolates carried two or three types of ESBLs, and producing CTX-M-14 and CTX-M-3 together were the most common type. CONCLUSION: The resistance of E. coli and K. pneumonia producing ESBLs in China was a serious issue and CTX-M type ESBLs were the most common genotype. CTX-M-14 was the predominant genotype. Some isolates produced two or three ESBLs.

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