Transferable DHA-1 cephalosporinase in Escherichia coli

Giakkoupi, Panagiota; Tambić-Andrašević, Arjana; Vourli, S.; Škrlin, Jasenka; Šestan-Crnek, Sandra; Tzouvelekis, L. S.; Vatopoulos, Alkiviadis

doi:10.1016/j.ijantimicag.2005.09.013

Cited by 16 publications

(10 citation statements)

References 14 publications

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“…This plasmid- mediated β-lactamase is now being increasingly detected in a strain of E. coli worldwide [31–33] and early detection of this β-lactamase (DHA-1) is mandatory for better antibiotic therapy and also to prevent further spread. The present study reports the prevalence of DHA-1 (9%) among E. coli strains in this region which is quite high compared to other studies [30, 32, 33] and typing of these DHA-1 harboring isolates by ERIC PCR revealed diverse haplotypes, indicating the spread of the DHA-1 gene through horizontal transfer. Based on the present susceptibility data (Table 2) and previous studies [11, 12], carbapenem and other non-β-lactam antibiotics such as tigecycline could be better drugs of choice for the treatment of infections caused by E. coli producing DHA-1.…”

Section: Discussioncontrasting

confidence: 49%

Occurrence of bla DHA-1 mediated cephalosporin resistance in Escherichia coli and their transcriptional response against cephalosporin stress: a report from India

Ingti

Paul

Maurya

et al. 2017

Ann Clin Microbiol Antimicrob

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BackgroundTreatment alternatives for DHA-1 harboring strains are challenging as it confers resistance to broad spectrum cephalosporins and may further limit treatment option when expressed at higher levels. Therefore, this study was designed to know the prevalence of DHA genes and analyse the transcription level of DHA-1 against different β-lactam stress.MethodsScreening of AmpC β-lactamase phenotypically by modified three dimensional extract method followed by Antimicrobial Susceptibility and MIC determination. Genotyping screening of β-lactamase genes was performed by PCR assay followed by their sequencing. The bla DHA-1 transcriptional response was evaluated under different cephalosporin stress by RT PCR. Transferability of bla DHA gene was performed by transformation and conjugation and plasmid incompatibility typing, DNA fingerprinting by enterobacterial repetitive intergenic consensus sequences PCR.Results16 DHA-1 genes were screened positive from 176 Escherichia coli isolates and primer extension analysis showed a significant increase in DHA-1 mRNA transcription in response to cefotaxime at 8 µg/ml (6.99 × 102 fold), ceftriaxone at 2 µg/ml (2.63 × 103 fold), ceftazidime at 8 µg/ml (7.06 × 103 fold) and cefoxitin at 4 µg/ml (3.60 × 104 fold) when compared with untreated strain. These transcription data were found significant when analyzed statistically using one way ANOVA. Four different ESBL genes were detected in 10 isolates which include CTX-M (n = 6), SHV (n = 4), TEM (n = 3) and OXA-10 (n = 1), whereas, carbapenemase gene (NDM) was detected only in one isolate. Other plasmid mediated AmpC β-lactamases CIT (n = 9), EBC (n = 2) were detected in nine isolates. All DHA-1 genes detected were encoded in plasmid and incompatibility typing from the transformants indicated that the plasmid encoding bla DHA-1 was carried mostly by the FIA and L/M Inc group.ConclusionThis study demonstrates the prevalence of DHA-1 gene in this region and highlights high transcription of DHA-1 when induced with different β-lactam antibiotics. Therefore, cephalosporin treatment must be restricted for the patients infected with pathogen expressing this resistance determinant.

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Section: Discussioncontrasting

confidence: 49%

Occurrence of bla DHA-1 mediated cephalosporin resistance in Escherichia coli and their transcriptional response against cephalosporin stress: a report from India

Ingti

Paul

Maurya

et al. 2017

Ann Clin Microbiol Antimicrob

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“…Antimicrobial susceptibility was determined by the broth micro-dilution method according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI, 2009a Carbapenemase genes (encoding class B families IMP, VIM, GIM, SPM and SIM; class A families NMC, IMI, SME, KPC and GES; and class D family OXA-48) (Queenan & Bush, 2007), plasmid-borne ampC-like genes (encoding CMY, DHA and ACT) (Alvarez et al, 2004;Chia et al, 2010;Giakkoupi et al, 2006) and ESBL genes (encoding CTX-M, Eckert et al, 2004;TEM, Eckert et al, 2004;and SHV, Rasheed et al, 1997) were detected by PCR amplification. Primers for the class B carbapenemase NDM-1 were newly designed in this study (NDM-1F: 59-TCTCGACATGCCGGGTTT-39; NDM-1R: 59-GAGATTGCCGAGCGACTT-39).…”

Section: Methodsmentioning

confidence: 99%

Molecular typing and resistance mechanisms of imipenem-non-susceptible Klebsiella pneumoniae in Taiwan: results from the Taiwan surveillance of antibiotic resistance (TSAR) study, 2002–2009

Siu

et al. 2013

Journal of Medical Microbiology

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We investigated the molecular mechanisms and clonality of imipenem-non-susceptible Klebsiella pneumoniae isolates collected during a Taiwan national surveillance programme, between 2002 and 2009. Genes for carbapenemases, plasmid-borne ampC-type genes and extended-spectrum b-lactamase (ESBL) genes were analysed by PCR. The major porin channels OmpK35 and OmpK36 were studied by SDS-PAGE. Molecular typing was performed with pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Our study revealed that all 29 of the isolates tested were ESBL producers. Of the K. pneumoniae isolates collected in Taiwan from 2002 to 2009, most (84.6 %, 11/13) imipenem-resistant (MIC .2 mg l "1 ) isolates carried the bla IMP-8 gene. Isolates with an imipenem MIC of 2 mg l "1 produced ESBLs with or without DHA-1 in combination with OmpK35/36 loss. PFGE analysis revealed that six small clusters of isolates were clonally related. The MLST grouping results were in concordance with the PFGE results. The predominant sequence types (ST) were ST11, ST48 and ST101. Two novel STs, ST1033 and ST1034, were found. The dominant clone in Taiwan, ST11, has been reported worldwide to be associated with various resistance mechanisms.

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“…The primers used in this study were described previously (1,5,11,12,14,18,19). PCR products were purified with a QIAquick PCR purification kit (Qiagen) and were sequenced on an ABI PRISM 3730XL sequencer analyzer.…”

mentioning

confidence: 99%

Phenotypic and Genotypic Characterization of Enterobacteriaceae with Decreased Susceptibility to Carbapenems: Results from Large Hospital-Based Surveillance Studies in China

Yang

Wang

Sun

et al. 2010

Antimicrob Agents Chemother

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The resistance mechanism of 49 Enterobacteriaceae isolates with decreased susceptibility to carbapenems collected from 2004 to 2008 at 16 teaching hospitals in China was investigated. Moderate-to high-level carbapenem resistance in most isolates was more closely associated with loss or decreased expression of both major porins combined with production of AmpC or extended-spectrum ␤-lactamase enzymes, while KPC-2, IMP-4, and IMP-8 carbapenemase production may lead to a low to moderate level of carbapenem resistance in Enterobacteriaceae in China.To date, the emergence of carbapenem-resistant Enterobacteriaceae has been reported in some countries (7,9,16,19). Carbapenemases and porin loss combined with AmpC enzyme hyperproduction are regarded as the main mechanisms of resistance (7,9,12,19). In China, there have been some reports of KPC-2-producing carbapenem-resistant Klebsiella pneumoniae, Serratia marcescens, and Escherichia coli in the city of Hangzhou (2,17,20). However, a nationwide survey has not been performed. In this study, 49 Enterobacteriaceae isolates with decreased susceptibility to carbapenems (MIC of imipenem, meropenem, or ertapenem of Ն2 g/ml) were collected from 16 teaching hospitals in a nationwide distribution, which included 26 K. pneumoniae, 8 E. coli, 10 Enterobacter cloacae, 2 Enterobacter aerogenes, and 3 Citrobacter freundii isolates. Identification of organisms was confirmed by using the API 20E or Vitek2 Compact system (bioMérieux, France). Susceptibility testing was performed by using the agar dilution method according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI) (3, 4). Breakpoints for tigecycline were as defined by the FDA (susceptible, Յ2 g/ml; resistant, Ն8 g/ml). Forty-nine isolates were nonsusceptible to most antibiotics except to tigecycline (to which 45 of 49 isolates were susceptible) and polymyxin B (to which 47 of 49 isolates were susceptible).Conjugation experiments were carried out in mixed broth cultures, as described previously (2). Plasmid DNAs of all carbapenemase-producing isolates were obtained with a QIAfilter midikit (Qiagen, Hilden, Germany). Resistance genes were successfully transferred from 23 of 49 isolates to the recipient E. coli C600. Among the 16 carbapenemaseproducing clinical isolates, carbapenemase genes of 13 isolates were successfully transferred to E. coli C600, except for three IMP-4-producing E. cloacae isolates. The 13 carbapenemaseproducing transconjugants showed 8-to 64-fold increases in the MIC of imipenem, 32-to 512-fold increases in the MIC of meropenem, and 256-to 4,096-fold increases in the MIC of ertapenem relative to those of the recipient. Most of the carbapenemase-producing transconjugants harbored a single plasmid, while only one transconjugant (GZ64T) harbored four different plasmids (Table 1).PCR of ␤-lactamase genes for the transconjugants and respective donors was carried in a PTC-200 PCR system (BioRad). The primers used in this study were described previously (1,5,11,12,14,18,19). PCR products were...

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Transferable DHA-1 cephalosporinase in Escherichia coli

Cited by 16 publications

References 14 publications

Occurrence of bla DHA-1 mediated cephalosporin resistance in Escherichia coli and their transcriptional response against cephalosporin stress: a report from India

Occurrence of bla DHA-1 mediated cephalosporin resistance in Escherichia coli and their transcriptional response against cephalosporin stress: a report from India

Molecular typing and resistance mechanisms of imipenem-non-susceptible Klebsiella pneumoniae in Taiwan: results from the Taiwan surveillance of antibiotic resistance (TSAR) study, 2002–2009

Phenotypic and Genotypic Characterization of Enterobacteriaceae with Decreased Susceptibility to Carbapenems: Results from Large Hospital-Based Surveillance Studies in China

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