Clinical Isolates Of Enterobacteriaceae Research Articles

Presence of qnr gene in Escherichia coli and Klebsiella pneumoniae resistant to ciprofloxacin isolated from pediatric patients in China

BackgroundQuinolone resistance in Enterobacteriaceae results mainly from mutations in type II DNA topoisomerase genes and/or changes in the expression of outer membrane and efflux pumps. Several recent studies have indicated that plasmid-mediated resistance mechanisms also play a significant role in fluoroquinolone resistance, and its prevalence is increasing worldwide. In China, the presence of the qnr gene in the clinical isolates of Enterobacteriaceae has been reported, but this transmissible quinolone resistance gene has not been detected in strains isolated singly from pediatric patients. Because quinolones associated with a variety of adverse side effects on children, they are not authorized for pediatric use. This study therefore aimed to investigate the presence of the qnr gene in clinical isolates of E. coli and K. pneumoniae from pediatric patients in China.MethodsA total 213 of non-repetitive clinical isolates resistant to ciprofloxacin from E. coli and K. pneumoniae were collected from hospitalized patients at five children's hospital in Beijing, Shanghai, Guangzhou, and Chongqing. The isolates were screened for the plasmid-mediated quinolone resistance genes of qnrA, qnrB, and qnrS by PCR. Transferability was examined by conjugation with the sodium azide-resistant E. coli J53. All qnr-positive were analyzed for clonality by enterobacterial repetitive intergenic consensus (ERIC)-PCR.ResultsThe study found that 19 ciprofloxacin-resistant clinical isolates of E. coli and K. pneumoniae were positive for the qnr gene, and most of the qnr positive strains were ESBL producers. Conjugation experiments showed that quinolone resitance could be transferred to recipients. Apart from this, different DNA banding patterns were obtained by ERIC-PCR from positive strains, which means that most of them were not clonally related.ConclusionThis report on transferable fluoroquinolone resistance due to the qnr gene among E. coli and K. pneumoniae strains indicated that plasmid-mediated quinolone resistance has emerged in pediatric patients in China.

Direct Comparison of the BD Phoenix System with the MicroScan WalkAway System for Identification and Antimicrobial Susceptibility Testing of Enterobacteriaceae and Nonfermentative Gram-Negative Organisms

The Phoenix automated microbiology system (BD Diagnostics, Sparks, MD) is designed for the rapid identification (ID) and antimicrobial susceptibility testing (AST) of clinically significant human bacterial pathogens. We evaluated the performance of the Phoenix instrument in comparison with that of the MicroScan WalkAway system (Dade Behring, West Sacramento, CA) in the ID and AST of gram-negative clinical strains and challenge isolates of Enterobacteriaceae (n = 150) and nonfermentative gram-negative bacilli (NFGNB; 45 clinical isolates and 8 challenge isolates). ID discrepancies were resolved with the API 20E and API 20NE conventional biochemical ID systems (bioMerieux, Durham, NC). The standard disk diffusion method was used to resolve discordant AST results. The overall percentages of agreement between the Phoenix ID results and the MicroScan results at the genus and species levels for clinical isolates of Enterobacteriaceae were 98.7 and 97.7%, respectively; following resolution with conventional biochemical testing, the accuracy of the Phoenix system was determined to be 100%. For NFGNB, the levels of agreement were 100 and 97.7%, respectively. Both systems incorrectly identified the majority of the uncommon nonfermentative nonpseudomonal challenge isolates recovered from cystic fibrosis patients; these isolates are not included in the databases of the respective systems. For AST of Enterobacteriaceae, the rate of complete agreement between the Phoenix results and the MicroScan results was 97%; the rates of very major, major, and minor errors were 0.3, 0.2, and 2.7%, respectively. For NFGNB, the rate of complete agreement between the Phoenix results and the MicroScan results was 89.1%; the rates of very major, major, and minor errors were 0, 0.5, and 7.7%, respectively. Following the confirmatory testing of nine clinical isolates initially screened by the MicroScan system as possible extended-spectrum-beta-lactamase (ESBL)-producing organisms (seven Klebsiella pneumoniae isolates and two Escherichia coli isolates), complete agreement was achieved for eight isolates (one ESBL positive and seven negative); one false positive was obtained with the Phoenix instrument. The MicroScan system correctly detected the 10 ESBL challenge isolates, versus the 6 detected by the Phoenix system. Overall, there was good correlation between the Phoenix instrument and the MicroScan system for the ID and AST of Enterobacteriaceae and common NFGNB. The Phoenix system is a reliable method for the ID and AST of the majority of clinical strains encountered in the clinical microbiology laboratory. Until additional performance data are available, results for all Klebsiella pneumoniae or Klebsiella oxytoca and E. coli isolates screened and confirmed as ESBL producers by any automated system should be confirmed by alternate methods prior to the release of final results.

Coprevalence of Plasmid-Mediated Quinolone Resistance Determinants QepA, Qnr, and AAC(6′)-Ib-cr among 16S rRNA Methylase RmtB-Producing Escherichia coli Isolates from Pigs

Plasmid-mediated quinolone resistance determinants, including Qnr peptides and AAC(6′)-Ib-cr, are increasingly identified worldwide among various clinical isolates of Enterobacteriaceae ([7][1], [9][2], [10][3]). Very recently, a novel plasmid-mediated fluoroquinolone-resistant determinant, QepA (

Prevalence and characterization of extended-spectrum β-lactamase–producing Enterobacteriaceae isolated in Korean hospitals

Prevalence and characteristics of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in Korean hospitals were assessed. A total of 1484 clinical Enterobacteriaceae isolates were collected from 8 tertiary-care hospitals in various regions of Korea over a 3-month period (June to August) in 2005. Among 546 Klebsiella pneumoniae isolates, 123 isolates (22.4%) showed ESBL-producing activity, and 47 (10.2%) of 460 isolates of Escherichia coli were ESBL producers. Of the Enterobacter cloacae isolates, 16.2% (17/105) evidenced ESBL-producing activity. The most prevalent ESBLs were SHV-12 and CTX-M-14 in K. pneumoniae and E. coli, respectively. In E. cloacae, SHV-12 was also the most prevalent. Prevalence of ESBL production differed among the specimens. Although the K. pneumoniae isolates from urine and aspirates evidenced high ESBL production rates (35.4% and 57.1%, respectively), those from sputum, blood, and pus showed relatively low ESBL production rates (17.0%, 14.8%, and 5.3%, respectively). However, E. coli isolates obtained from sputum showed significantly higher ESBL production rates (37.5%) than were seen in samples obtained from other sources, but those obtained from urine showed lower ESBL production rates (8.3%). These significant differences in ESBL-producing K. pneumoniae and E. coli isolates among the isolated specimens should be examined further, with an eye toward the implications of this research in clinical settings.

Surveillance for plasmid-mediated quinolone resistance determinants in Enterobacteriaceae within the Calgary Health Region, Canada: the emergence of aac(6')-Ib-cr

A study was designed to determine the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants among clinical isolates of Enterobacteriaceae from the Calgary Health Region (CHR). During January and February 2004 and January and February 2007, 564 non-repeat isolates of Escherichia coli, Klebsiella spp., Proteus mirabilis and Morganella morganii resistant to ciprofloxacin and/or tobramycin were screened for PMQR determinants using a multiplex PCR for qnrA, qnrS and qnrB and aac(6')-Ib genes; aac(6')-Ib-cr was further identified by digestion with BstF5I. In 2004, 6/139 (4%) of the resistant E. coli were positive for aac(6')-Ib-cr. In 2007, 53/398 (13%) were positive for aac(6')-Ib-cr, 3/398 (0.8%) were positive for qnrS, and one isolate was positive for both. All the isolates were present in urines and the majority [40/63 (63%)] were submitted from community collection sites; 8 (13%) isolates co-produced AmpC beta-lactamases and 34 (54%) co-produced CTX-M-15. aac(6')-Ib-cr was present in one Klebsiella pneumoniae and one P. mirabilis, whereas one isolate of K. pneumoniae was positive for both aac(6')-Ib-cr and qnrB. Our results showed that isolates with aac(6')-Ib-cr, often associated with CTX-M-15, are emerging among fluoroquinolone-resistant E. coli in the CHR. Our study suggests that surveillance for PMQR determinants should be undertaken on a regular basis.

Multiplex PCR amplification assay for the detection of blaSHV, blaTEM and blaCTX‐M genes in Enterobacteriaceae

Extended-spectrum beta-lactamases (ESBLs) are often mediated by (bla-)SHV, (bla)TEM and (bla)CTX-M genes in Enterobacteriaceae and other Gram-negative bacteria. Numerous molecular typing methods, including PCR-based assays, have been developed for their identification. To reduce the number of PCR amplifications needed we have developed a multiplex PCR assay which detects and discriminates between (bla-)SHV, (bla)TEM and (bla)CTX-M PCR amplicons of 747, 445 and 593 bp, respectively. This multiplex PCR assay allowed the identification of (bla-)SHV, (bla)TEM and (bla)CTX-M genes in a series of clinical isolates of Enterobacteriaceae with previously characterised ESBL phenotype. The presence of (bla)SHV, (bla)TEM and (bla)CTX-M genes was confirmed by partial DNA sequence analysis. Apparently, the universal well-established CTX-M primer pair used here to reveal plasmid-encoded (bla)CTX-M genes would also amplify the chromosomally located K-1 enzyme gene in all Klebsiella oxytoca strains included in the study.

Characterization of sulphonamide resistance genes and class 1 integron gene cassettes in Enterobacteriaceae, Central African Republic (CAR)

The aim of this study was to characterize genes encoding sulphonamide resistance and gene cassettes associated with class 1 integrons in trimethoprim-sulphamethoxazole resistant Enterobacteriaceae recovered from Bangui, Central African Republic (CAR). We studied 78 clinical Enterobacteriaceae isolates, including 16 extended-spectrum beta-lactamases producers, 10 Salmonella and 9 Shigella, resistant to trimethoprim-sulphamethoxazole as assessed by the disc diffusion method. PCR was used to test for sul1 and sul2 genes. Class 1 integron resistance gene cassettes were characterized by directly sequencing PCR products obtained with primers recognising 5' and 3' conserved regions. The sul1 gene was found in 67 isolates, the sul2 gene in 72 isolates and both genes in 62 isolates, while the int1 gene was found in 74 isolates. The most prevalent dfr genes were dfrA7 (49%), dfrA1 (17%) and dfrA2d (13%). These results illustrate the wide distribution of sulphonamide and trimethoprim resistance genes among Enterobacteriaceae in Bangui (CAR).

P1026 Occurrence and diversity of genetic resistance determinants among Enterobacteriaceae clinical isolates in Aveiro, Portugal

P1026 Occurrence and diversity of genetic resistance determinants among Enterobacteriaceae clinical isolates in Aveiro, Portugal

Antimicrobial activity of Uncaria tomentosa against oral human pathogens

Uncaria tomentosa is considered a medicinal plant used over centuries by the peruvian population as an alternative treatment for several diseases. Many microorganisms usually inhabit the human oral cavity and under certain conditions can become etiologic agents of diseases. The aim of the present study was to evaluate the antimicrobial activity of different concentrations of Uncaria tomentosa on different strains of microorganisms isolated from the human oral cavity. Micropulverized Uncaria tomentosa was tested in vitro to determine the minimum inhibitory concentration (MIC) on selected microbial strains. The tested strains were oral clinical isolates of Streptococcus mutans, Staphylococcus spp., Candida albicans, Enterobacteriaceae and Pseudomonas aeruginosa. The tested concentrations of Uncaria tomentosa ranged from 0.25-5% in Müeller-Hinton agar. Three percent Uncaria tomentosa inhibited 8% of Enterobacteriaceae isolates, 52% of S. mutans and 96% of Staphylococcus spp. The tested concentrations did not present inhibitory effect on P. aeruginosa and C. albicans. It could be concluded that micropulverized Uncaria tomentosa presented antimicrobial activity on Enterobacteriaceae, S. mutans and Staphylococcus spp. isolates.

P1036 Clinical urinary tract isolates of Enterobacteriaceae in the Arkhangelsk region, Russian Federation: antimicrobial resistance profiles and characterisation of ESBL-strains

P1036 Clinical urinary tract isolates of Enterobacteriaceae in the Arkhangelsk region, Russian Federation: antimicrobial resistance profiles and characterisation of ESBL-strains

P1538 The severity of Streptococcus pyogenes infections in children is significantly associated with plasma levels of infiammatory cytokines

The susceptibility testing accuracy of the VITEK2 system and the ability of the Advance Expert System (AES) to provide interpretive readings were evaluated against 86 extended spectrum (ESBL) and 6 inhibitor-resistant-TEM (IRT) β-lactamases producing Enterobacteriaceae clinical isolates. VITEK2 MICs of 12 β-lactams were compared with those obtained by the standard NCCLS microdilution technique. The overall essential agreement ( ± 1 log dilution) was 87.8%. Discrepancies were mainly observed with cefepime (30.3% of total number of discrepancies), ceftazidime (21.2%), and cefotaxime (15.1%). MIC discrepancies were slightly higher in CTX-M- (14.4%) than in TEM- (12.5%) or SHV- (11.9%) type ESBL producers and were rare in IRT producers (1.4%). Overall interpretive agreement was 92.5% and minor, major, and very major errors were 5.4%, 1.7%, and 2.1%, respectively. The AES was able to identify an ESBL phenotype in 85 out of 86 isolates (98.8%) and an IRT phenotype in all 6 isolates harboring these enzymes, thus reducing very major errors to 0.9%. The VITEK2 system, in conjunction with the AES software, is a reliable tool for detection of ESBL or IRT producing Enterobacteriaceae isolates.

Emergence of ArmA and RmtB aminoglycoside resistance 16S rRNA methylases in Belgium

16S rRNA methylase-mediated high-level resistance to aminoglycosides has been reported recently in clinical isolates of Gram-negative bacilli only from a limited number of countries. This study was conducted to investigate the occurrence of this type of resistance in clinical isolates of Enterobacteriaceae from two Belgian hospitals and the characteristics of the strains. We screened for high-level gentamicin, tobramycin and amikacin resistance in clinical isolates of Enterobacteriaceae consecutively collected between 2000 and 2005 at two laboratories by PCR for the armA, rmtA and rmtB 16S rRNA methylase genes. The beta-lactamase presence in the strains was also determined by phenotypic and genotypic methods. Overall armA genes were detected in 18 Klebsiella pneumoniae, Escherichia coli, Enterobacter aerogenes, Enterobacter cloacae and Citrobacter amalonaticus whereas rmtB was detected in a single E. coli isolate. The rmtA gene was not found. All 16S rRNA methylase-bearing strains produced extended-spectrum beta-lactamases (ESBLs), predominantly type CTX-M-3, as well as various types of beta-lactamases. In the majority of the strains, the armA gene was carried by conjugative plasmids of the IncL/M incompatibility group whereas rmtB was borne by an IncFI plasmid. This is the first report of the emergence of 16S rRNA methylases in Enterobacteriaceae in Belgium. The rapid spread of multidrug-resistant isolates producing both ESBLs and 16S rRNA methylases raises clinical concern and may become a major therapeutic threat in the future.

Occurrence of qnrA-positive clinical isolates in French teaching hospitals during 2002–2005

Bacteria harbouring the novel qnrA plasmid-mediated mechanism of quinolone resistance have been described in different countries, but the frequency of their occurrence has not been investigated. In total, 1,468 clinical isolates of Enterobacteriaceae with quinolone resistance or extended-spectrum beta-lactamase (ESBL) phenotypes were collected from eight teaching hospitals in France during 2002-2005 and screened for qnrA. Overall, 28 isolates (22 Enterobacter cloacae, three Klebsiella pneumoniae, one Citrobacter freundii, one Klebsiella oxytoca and one Proteus mirabilis) were positive for qnrA, representing 1.9% of all isolates, 3.3% of ESBL-producing isolates (22% of the E. cloacae isolates) and 0% of non-ESBL-producing isolates. The prevalence of qnrA among consecutive ESBL-producing isolates in 2004 from the eight hospitals was 2.8% (18/639). Of the qnrA-positive isolates, 100% were intermediately-resistant or resistant to nalidixic acid, and 75% to ciprofloxacin. Twenty-one of the 22 qnrA-positive E. cloacae isolates were obtained from two hospitals in the Paris area, and molecular typing and plasmid content analysis showed clonal relationships for five, three and two isolates, respectively. The qnrA genetic environment was similar to that of the In36 integron. The remaining two isolates had qnrA variants (30 and 29 nucleotide differences, respectively, compared with the original sequence) and an unknown genetic environment. The ESBL gene associated with qnrA was bla(SHV-12) in most of the isolates, but bla(PER-1) and bla(SHV-2a) were found in two isolates. In France, it appears that qnrA-positive isolates are predominantly E. cloacae isolates producing SHV-12, and may be associated with the dissemination of an In36-like integron.

Rapid identification of Enterobacteriaceae by sequencing DNA gyrase subunit B encoding gene

Real-time polymerase chain reaction and sequencing were used to characterize a 506-bp-long DNA fragment internal to the gyrB gene ( gyrBint). The sequences obtained from 32 Enterobacteriaceae-type strains and those available in the Genbank nucleotide sequence database ( n = 24) were used as a database to identify 240 clinical enterobacteria isolates. Sequence analysis of the gyrBint fragment of 240 strains showed that gyrBint constitutes a discriminative target sequence to differentiate between Enterobacteriaceae species. Comparison of these identifications with those obtained by phenotypic methods (Vitek 1 system and/or Rapid ID 32E; bioMérieux, Marcy l'Etoile, France) revealed discrepancies essentially with genera Citrobacter and Enterobacter. Most of the strains identified as Enterobacter cloacae by phenotypic methods were identified as Enterobacter hormaechei strains by gyrBint sequencing. The direct sequencing of gyrBint would be useful as a complementary tool in the identification of clinical Enterobacteriaceae isolates.

First report of CTX-M-15 and CTX-M-3 β-lactamases among clinical isolates of Enterobacteriaceae in Béjaia, Algeria

We assessed the prevalence and phenotypic characteristics of extended-spectrum β-lactamase producers among 365 clinical isolates of members of the family Enterobacteriaceae recovered from two hospitals in Béjaia, Algeria, between March 2004 and April 2005. Twenty-one strains were resistant to cefotaxime and/or ceftazidime. A double-disk synergy test yielded a positive result in five cases (three Escherichia coli, one Klebsiella pneumoniae and one Enterobacter cloacae). Using polymerase chain reaction and sequencing, the three E. coli isolates and the K. pneumoniae isolate were found to produce extended-spectrum β-lactamase CTX-M-15 and the E. cloacae isolate produced CTX-M-3. The three CTX-M-15-producing E. coli isolates were not isolated in the same wards, although genotyping by randomly amplified polymorphic DNA analysis revealed that they were clonally related. The bla CTX-M-15 genes were transferred from E. coli by conjugation, whilst conjugative transfer of bla CTX-M genes from K. pneumoniae and E. cloacae was not detectable.

Spread of blaCTX-M-type and blaPER-2 β-lactamase genes in clinical isolates from Bolivian hospitals

To assess the prevalence and types of genes encoding extended-spectrum beta-lactamases (ESBLs) in clinical isolates of Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter spp. from Bolivia. A total of 642 clinical isolates were collected consecutively during a 4 month period (September to December 2004). Resistance or reduced susceptibility to cefotaxime and/or ceftazidime and/or aztreonam was assessed using double disc synergy tests using clavulanic acid, cefotaxime, ceftazidime and aztreonam to identify putative ESBL-producing isolates. The ESBL determinants were characterized by colony blot hybridization, PCR and DNA sequencing. Of the 642 isolates, 220 (34.3%) showed resistance or reduced susceptibility to cefotaxime and/or ceftazidime and/or aztreonam, and 150 (23.4%) were putative ESBL producers. A total of 106 ESBL-producing isolates contained the bla(CTX-M-2) gene, and 32 isolates had a novel allele, bla(CTX-M-43). bla(CTX-M) alleles were detected in all P. aeruginosa and Acinetobacter spp. studied. In contrast, only 12 ESBL-producing isolates had bla(PER-2), mainly Enterobacteriaceae, although it was also found in a strain of P. aeruginosa. This is the first study on ESBL-producing strains in Bolivia and it reveals a high prevalence of bla(CTX-M) genes. The PER-2 enzyme was less prevalent, but its gene was detected in several species, including P. aeruginosa, which is consistent with horizontal transfer.

Accelerated detection of extended-spectrum β-lactamases in clinical isolates of Enterobacteriaceae

A prospective study is carried out to evaluate the performance of a protocol for the accelerated detection of extended-spectrum β-lactamases (ESBLs) in clinical isolates of Escherichia coli, Klebsiella pneumoniae and other Gram-negative bacteria. A modified double-disc test (MDDT) is incorporated in a Gram-negative template for routine susceptibility testing. The MDDT identified accurately ESBLs in all isolates subsequently confirmed as ESBLproducers by the standard Clinical Laboratory Standards Institute (CLSI) combined disc method. Of 1213 isolates tested, 98 (8%) were positive for ESBLs by MDDT and 95 (7.8%) were positive by the CLSI method. ESBLs were detected in 48 (7.8%) E. coli, 21 (8%) K. pneumoniae, 12 (5.8%) Proteus mirabilis, 13 (18.8%) Providencia stuartii and four (6.8%) Enterobacter cloacae isolates. Time required for ESBL detection by the MDDT method was one day. The protocol described provides a simple, rapid and low-cost method for early detection of ESBLs in Gram-negative bacteria.

Comparación de tres métodos microbiológicos para la detección de betalactamasas de espectro extendido en enterobacterias aisladas en Santa Fe (Argentina)

Expanded-spectrum betalactamases (ESBLs) are the main source of resistance to oxyimino cephalosporins and monobactams in Enterobacteriaceae. Most of them derive from TEM or SHV, however the incidence of other families like CTX-M, OXA and PER has increased. In Argentina, the most frequent ESBL in Enterobacteriaceae is CTX-M-2. This specific circumstance, which differs from the situation in the Northern Hemisphere, motivated us to study new diagnostic strategies for the detection of ESBLs in our region. Microbiological ESBL detection was performed by double-disk synergy tests, cefotaxime and ceftazidime disks with and without clavulanic acid (NCCLS), and cefotaxime and ceftazidime disks in Müeller-Hinton agar supplemented with lithium clavulanate (MH-cla). Betalactamases were characterized by isoelectric focusing, hydrolysis profile and PCR amplification. Among 575 clinical isolates of Enterobacteriaceae, 14% were oxyimino cephalosporin-resistant. Two different ESBLs were detected in 31 resistant strains: CTX-M-2 (28) and PER-2 groups (3). The double-disk synergy test was the least sensitive method for ESBL detection. ESBLs were detected by the other two methods in all isolates with the use of cefotaxime disks, but not with ceftazidime disks. The microbiological method employing MH-cla with cefotaxime disks had a sensitivity and specificity comparable to the referral test using the same antibiotic proposed by the NCCLS for the detection of ESBLs.

Dissemination of transferable AmpC-type beta-lactamase (CMY-10) in a Korean hospital.

To determine dissemination and genotype of AmpC beta-lactamases and an extended-spectrum beta-lactamase among clinical isolates of Enterobacteriaceae, we performed antibiotic susceptibility testing, pI determination, induction test, plasmid profiles, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Among the 51 clinical isolates collected from a university hospital in Korea, six isolates were resistant to cephamycins. All six isolates produced a plasmid-encoded AmpC-type beta-lactamase, CMY-10. Five strains also produced one or more other beta-lactamases: SHV-12, an extended-spectrum beta-lactamase (five isolates); TEM-1, a class A beta-lactamase (two isolates); and a chromosomal AmpC beta-lactamase (one isolate, a strain of Enterobacter aerogenes, which produced all four of the beta-lactamases that were identified). One of six isolates produced only CMY-10. ERIC-PCR analysis revealed that dissemination of CMY-10 and SHV-12 was due to a clonal outbreak of a resistant strain and to the interspecies spread of resistance to cephamycins and broad-spectrum beta-lactams in Korea. CMY-10 beta-lactamase genes that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized from six clinical isolates. A sequence identical to the common regions in In6, In7, and a novel integron from pSAL-1 was found upstream from blaCMY-10 gene at nucleotides 1-71. A total of 15 nucleotides (I-15) or 18 nucleotides (I-18) between position 71 and 72 were inserted into the blaCMY-10 gene. The blaCMY-10 gene might be inserted into a sul1-type complex integron by I-15 or I-18.

Dissemination of Transferable AmpC-type β-Lactamase (CMY-10) in a Korean Hospital

To determine dissemination and genotype of AmpC β-lactamases and an extended-spectrum β-lactamase among clinical isolates of Enterobacteriaceae, we performed antibiotic susceptibility testing, pI determination, induction test, plasmid profiles, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Among the 51 clinical isolates collected from a university hospital in Korea, six isolates were resistant to cephamycins. All six isolates produced a plasmid-encoded AmpC-type β-lactamase, CMY-10. Five strains also produced one or more other β-lactamases: SHV-12, an extended-spectrum β-lactamase (five isolates); TEM-1, a class A β-lactamase (two isolates); and a chromosomal AmpC β-lactamase (one isolate, a strain of Enterobacter aerogenes, which produced all four of the β-lactamases that were identified). One of six isolates produced only CMY-10. ERIC-PCR analysis revealed that dissemination of CMY-10 and SHV-12 was due to a clonal outbreak of a resistant strain and to the interspecies spread of resistance to cephamycins and broad-spectrum β-lactams in Korea. CMY-10 β-lactamase genes that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized from six clinical isolates. A sequence identical to the common regions in In6, In7, and a novel integron from pSAL-1 was found upstream from blaCMY-10 gene at nucleotides 1–71. A total of 15 nucleotides (I-15) or 18 nucleotides (I-18) between position 71 and 72 were inserted into the blaCMY-10 gene. The blaCMY-10 gene might be inserted into a sul1-type complex integron by I-15 or I-18.