Detection of Five Novel CTX-M-Type Extended Spectrum Beta-Lactamases with One to Three CTX-M-14 Point Mutations in Isolates from Hefei, Anhui Province, China

Li, Hui; Li, Jia Bin

doi:10.1128/jcm.43.8.4301-4302.2005

Cited by 9 publications

(5 citation statements)

References 9 publications

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“…1). CTX-M-14 was expected to be the most probable variant forming the middle hybrid part because the highest prevalence of this enzyme among the CTX-M-9 group of enzymes described above has been noted in the Far East (3,16,19,35). Thus, this novel CTX-M-type ␤-lactamase has been assigned the designation CTX-M-64 by G. A. Jacoby (http://www .lahey.org/studies/webt.asp), which differed from the CTX-M-15-like ␤-lactamase by 22 amino acid residues (92.4% similarity) and from the CTX-M-14 ␤-lactamase by 35 amino acid residues (88.0% similarity).…”

Section: Resultsmentioning

confidence: 99%

Novel Chimeric β-Lactamase CTX-M-64, a Hybrid of CTX-M-15-Like and CTX-M-14 β-Lactamases, Found in a Shigella sonnei Strain Resistant to Various Oxyimino-Cephalosporins, Including Ceftazidime

Nagano

Wachino

et al. 2009

Antimicrob Agents Chemother

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The plasmid-mediated novel ␤-lactamase CTX-M-64 was first identified in Shigella sonnei strain UIH-1, which exhibited resistance to cefotaxime (MIC, 1,024 g/ml) and ceftazidime (MIC, 32 g/ml). The amino acid sequence of CTX-M-64 showed a chimeric structure of a CTX-M-15-like ␤-lactamase (N-and C-terminal moieties) and a CTX-M-14-like ␤-lactamase (central portion, amino acids 63 to 226), suggesting that it originated by homologous recombination between the corresponding genes. The introduction of a recombinant plasmid carrying bla CTX-M-64 conferred resistance to cefotaxime in Escherichia coli, and the activities of cefotaxime and ceftazidime were restored in the presence of clavulanic acid. Of note, CTX-M-64 production could also confer consistent resistance to ceftazidime, which differs from the majority of CTX-M-type enzymes, which poorly hydrolyze ceftazidime. These results were consistent with the kinetic parameters determined with the purified CTX- Shigellosis remains a public health concern throughout the world and has become an actual threat, particularly in developing countries, where 99% of the estimated 165 million annual episodes occur. Children under 5 years of age have been involved in more than half of the episodes and deaths (14). Shigellosis is more severe in malnourished children and elderly and immunocompromised people. Antibiotic treatment shortens the duration of clinical symptoms and pathogen excretion, prevents disease transmission, and reduces the risk of potential complications (18,27,34). However, empirical therapy with first-line antimicrobial agents, including ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol, nalidixic acid, co-trimoxazole, and tetracycline, has become less effective due to the high prevalence of multidrug-resistant (MDR) clinical isolates among Shigella species (9, 28, 29, 32). For these MDR isolates, the therapeutic options for oral administration are fluoroquinolones for adults and oxyimino-cephalosporins for children.Plasmid-encoded class A extended-spectrum ␤-lactamase (ESBL) production is still uncommon among Shigella species, despite the worldwide spread and prevalence of ESBL-producing clinical isolates belonging to the family Enterobacteriaceae. Four CTX-M-type ␤-lactamases, CTX-M-2, CTX-M-3, CTX-M-14, and CTX-M-15, and several TEM-derived ESBLs have been reported for Shigella sonnei (1,11,15,25). S. sonnei strain UIH-1, characterized in this study, produced a novel CTX-Mtype ␤-lactamase, a hybrid of the CTX-M-15-like ␤-lactamase, which is a new CTX-M-15 variant (GenBank accession no. DQ256091), and the CTX-M-14 ␤-lactamase; and this chimeric enzyme conferred resistance to ceftazidime as well as to cefotaxime and ceftriaxone. MATERIALS AND METHODSClinical isolate. S. sonnei UIH-1 was identified with the API 20E system (bioMérieux) in combination with tests for the utilization of citrate with Christensen's citrate medium (4), sodium acetate, and mucate and by PCR detection of the invE and ipaH genes with specific primer sets (Takara Bio, Shiga, Japan)...

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Section: Resultsmentioning

confidence: 99%

Novel Chimeric β-Lactamase CTX-M-64, a Hybrid of CTX-M-15-Like and CTX-M-14 β-Lactamases, Found in a Shigella sonnei Strain Resistant to Various Oxyimino-Cephalosporins, Including Ceftazidime

Nagano

Wachino

et al. 2009

Antimicrob Agents Chemother

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“…The presence of ␤-lactamases genes (ESBLs bla SHV , bla TEM , bla CTX-M-1 , bla CTX-M-2 , bla CTX-M-8 and bla CTX-M-9 and AmpC enzymes bla MOX-1 , bla MOX-2 , bla CMY-1-11, bla LAT-1-4 , bla BIL-1 , bla DHA-1 , bla DHA-2 , bla ACC , bla MIR-1T , bla ACT-1 , and bla FOX-1-5b ) was assessed on the qnrpositive isolates using specific primers [15][16][17][18].…”

Section: Polymerase Chain Reaction (Pcr) Amplification and Dna Sequenmentioning

confidence: 99%

Characterisation of qnr plasmid-mediated quinolone resistance in Enterobacteriaceae from Italy: association of the qnrB19 allele with the integron element ISCR1 in Escherichia coli

Richter

Frasson

Bergo

et al. 2010

International Journal of Antimicrobial Agents

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The spread of plasmid-mediated quinolone resistance determinants (qnr-like determinants) was evaluated in a collection of 232 ciprofloxacin-resistant or extended-spectrum β-lactamase (ESBL)-producing enterobacterial isolates recovered between November 2007 and May 2008 at Padua University Hospital, Italy. qnr genes were mainly found in Klebsiella pneumoniae (68%) and to a lesser extent in Escherichia coli (5.1%). Among the qnrA1, qnrS1 and qnrB19 alleles found, the latter was by far the most frequent. Genetic environment analysis revealed that one qnrB19 gene in E. coli was embedded in an ISCR1 complex class 1 integron. All other qnrB19 genes were flanked by an ISEcp1C region as part of the Tn2012 transposon. qnrA1- and qnrS1-containing strains were not clonally related. Both topoisomerase II mutations and ESBL (mainly SHV-12, TEM-1 and TEM-150 types) were present in most of the qnr-positive strains. qnrB19 is extremely frequent in K. pneumoniae isolates from Italy. In addition, association of qnrB19 with the ISCR1 mobile element in E. coli suggests a broad distribution of this resistance gene in the near future

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“…In our previous report, 5 strains of Klebsiella pneumoniae (K. pneumoniae) and 3 strains of E. coli produced 5 novel CTX-M enzymes were identified in our area [10] . These novel enzymes came from CTX-M-14 with 1−3 amino acid substitution by sequencing and the BLAST program (http:// www.ncbi.nlm.nih.gov/BLAST/).…”

Section: Introductionmentioning

confidence: 82%

“…Plasmid DNA extracted from 8 clinical isolates and their transconjugant by rapid alkaline lysis protocol was used as the template. PCR amplification was carried out under the conditions as previously described [10] . The purified PCR products were ligated with pGEM-Teasy vectors (Promega, Madison, Wisconsin, USA) and expressed in E. coli DH 5a .…”

Section: Methodsmentioning

confidence: 99%

Phenotypic and molecular characterization of 5 novel CTX-M enzymes carried by Klebsiella pneumoniae and Escherichia coli

Cheng

Yan

Wang

et al. 2008

Acta Pharmacologica Sinica

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Aim: The aim of the present study was to study the phenotypic and molecular characterization of 5 novel CTX-M-β-1actamases carried by 5 Klebsiella pneumoniae isolates and 3 Escherichia coli isolates collected from 4 hospitals in Hefei, China. Methods: The purified PCR products were ligated with pGEM-Teasy vectors, expressed, and sequenced. The complete genes of the CTX-M-β-lactamases were ligated with the pHSG398 vector to express prokaryotic recombinant proteins. Plasmids were extracted by rapid alkaline lysis protocol, and the PCR method was performed to determine whether the prokaryotic expression was successful or not. Antimicrobial susceptibility was tested and the phenotypes of transformants were determined according to criteria recommended by the Clinical and Laboratory Standards Institute. The kinetic parameters of enzymes were confirmed. The isoelectric points (pI) were determined by isoelectric focusing assay. Pulsed-field gel electrophoresis and plasmid profiling were performed. Results: The PCR products had 1101 nucleotides and were determined as CTX-M-46, CTX-M-47, CTX-M-48, CTX-M-49, and CTX-M-50. All strains were resistant to cefotaxime, but most of them were susceptible or intermediate to ceftazidime. The phenotypes of novel enzymes were determined as extended-spectrum β-lactamases (ESBL). Penicillin G, cephalothin, cefuroxime, and cefotaxime were determined to good substrates, whereas ceftazidime hydrolysis was not detected. The pI of the 5 novel CTX-M-β-lactamases were 8.0. CTX-M-derivatives could be the multiplex genesis in our area. Conclusion: This is the first report of these 5 novel plasmid-mediated CTX-M ESBL produced from China in the world. Molecular typing reveals notably different origin in genes encoding different CTX-M variants of 8 strains.

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Detection of Five Novel CTX-M-Type Extended Spectrum Beta-Lactamases with One to Three CTX-M-14 Point Mutations in Isolates from Hefei, Anhui Province, China

Cited by 9 publications

References 9 publications

Novel Chimeric β-Lactamase CTX-M-64, a Hybrid of CTX-M-15-Like and CTX-M-14 β-Lactamases, Found in a Shigella sonnei Strain Resistant to Various Oxyimino-Cephalosporins, Including Ceftazidime

Novel Chimeric β-Lactamase CTX-M-64, a Hybrid of CTX-M-15-Like and CTX-M-14 β-Lactamases, Found in a Shigella sonnei Strain Resistant to Various Oxyimino-Cephalosporins, Including Ceftazidime

Characterisation of qnr plasmid-mediated quinolone resistance in Enterobacteriaceae from Italy: association of the qnrB19 allele with the integron element ISCR1 in Escherichia coli

Phenotypic and molecular characterization of 5 novel CTX-M enzymes carried by Klebsiella pneumoniae and Escherichia coli

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