Molecular characterization of theblaKPC-2gene in clinical isolates of carbapenem-resistantKlebsiella pneumoniaefrom the pediatric wards of a Chinese hospital

2012 ◽  
Vol 58 (10) ◽  
pp. 1167-1173 ◽  
Author(s):  
Yang Liu ◽  
Xiang-Yang Li ◽  
La-Gen Wan ◽  
Wei-Yan Jiang ◽  
Fang-Qu Li ◽  
...  
Keyword(s):  
16S Rrna ◽  
Antibiotic Resistance Gene ◽  
Restriction Endonuclease Analysis ◽  
Multidrug Resistant ◽  
Class 1 Integron ◽  
Integrase Gene ◽  
Klebsiella Pneumoniae Carbapenemase ◽  
Class 1 ◽  
16S Rrna Methylase

The present study was conducted to confirm the presence of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae associated with a nosocomial outbreak in a Chinese pediatric hospital. From July 2009 to January 2011, 124 nonduplicated K. pneumoniae isolates were collected from specimens from patients of pediatric units in the hospital. Twelve of the 124 isolates possessed the blaKPC-2gene and showed 7 different pulsed-field gel electrophoresis (PFGE) patterns. Meanwhile, 16S rRNA methylase, acc(6′)-Ib-cr, and several types of β-lactamases were also produced by the majority of the KPC-producing isolates. Class 1 integron-encoded intI1 integrase gene was subsequently found in all strains, and amplification, sequencing, and comparison of DNA between 5′ conserved segment and 3′ conserved segment region showed the presence of several known antibiotic resistance gene cassettes of various sizes. The conjugation and plasmid-curing experiments indicated some KPC-2-encoding genes were transmissible. In addition, conjugal cotransfer of multidrug-resistant phenotypes with KPC-positive phenotypes was observed in KPC-producing strains. Restriction endonuclease analysis and DNA hybridization with a KPC-specific probe showed that the blaKPC-2gene was carried by plasmid DNA from K. pneumoniae of PFGE pattern B. The overall results indicate that the emergence and outbreak of KPC-producing K. pneumoniae in our pediatric wards occurred in conjunction with plasmids coharboring 16S rRNA methylase and extended-spectrum β-lactamases.

scholarly journals Molecular Characterization of β-Lactamase Genes and Their Genetic Structures in Acinetobacter Genospecies 3 Isolates in Taiwan

2010 ◽  
Vol 54 (6) ◽  
pp. 2699-2703 ◽  
Author(s):  
Li-Yueh Huang ◽  
Po-Liang Lu ◽  
Te-Li Chen ◽  
Fang-Yee Chang ◽  
Chang-Phone Fung ◽  
...  
Keyword(s):  
Genetic Structure ◽  
Molecular Characterization ◽  
Class 1 Integron ◽  
Integrase Gene ◽  
Class 1 ◽  
Genetic Structures

ABSTRACT The genetic structure of β-lactamases in Acinetobacter genospecies 3 (AG3) isolates in Taiwan was studied to analyze their high rates of resistance to β-lactams, including carbapenems (57.9%). bla IMP-1 and bla IMP-8 were located in a class 1 integron. bla OXA-58 was bracketed by ISAba3. A novel TnpF-like integrase gene was identified upstream of bla VEB-3. Adjacent to the 5′ sequence of the bla ADC gene, folE was identified. Four new Acinetobacter-derived cephalosporinase (ADC) enzymes were found, which clustered phylogenetically with published AG3 ADC proteins.

scholarly journals Characterization of In53, a Class 1 Plasmid- and Composite Transposon-Located Integron of Escherichia coli Which Carries an Unusual Array of Gene Cassettes

2001 ◽  
Vol 183 (1) ◽  
pp. 235-249 ◽  
Author(s):  
Thierry Naas ◽  
Yuzuru Mikami ◽  
Tamae Imai ◽  
Laurent Poirel ◽  
Patrice Nordmann
Keyword(s):  
Escherichia Coli ◽  
Resistance Gene ◽  
Gene Cassette ◽  
Promoter Sequence ◽  
Class 1 Integron ◽  
Integrase Gene ◽  
Chloramphenicol Resistance ◽  
Gene Cassettes ◽  
Class 1

ABSTRACT Further characterization of the genetic environment of the gene encoding the Escherichia coli extended-spectrum β-lactamase, bla VEB-1, revealed the presence of a plasmid-located class 1 integron, In53, which carried eight functional resistance gene cassettes in addition tobla VEB-1. While the aadB and the arr-2 gene cassettes were identical to those previously described, the remaining cassettes were novel: (i) a novel nonenzymatic chloramphenicol resistance gene of the cmlAfamily, (ii) a qac allele encoding a member of the small multidrug resistance family of proteins, (iii) a cassette,aacA1b/orfG, which encodes a novel 6′-N-acetyltransferase, and (iv) a fused gene cassette,oxa10/aadA1, which is made of two cassettes previously described as single cassettes. In addition, oxa10 andaadA1 genes were expressed from their own promoter sequence present upstream of the oxa10 cassette.arr-2 coded for a protein that shared 54% amino acid identity with the rifampin ADP-ribosylating transferase encoded by thearr-1 gene from Mycobacterium smegmatisDSM43756. While in M. smegmatis, the main inactivated compound was 23-ribosyl-rifampin, the inactivated antibiotic recovered from E. coli culture was 23-O-ADP-ribosyl-rifampin. The integrase gene of In53 was interrupted by an IS26 insertion sequence, which was also present in the 3′ conserved segment. Thus, In53 is a truncated integron located on a composite transposon, named Tn2000, bounded by two IS26 elements in opposite orientations. Target site duplication at both ends of the transposon indicated that the integron likely was inserted into the plasmid through a transpositional process. This is the first description of an integron located on a composite transposon.

scholarly journals Identification and Characterization of Class 1 Integron Resistance Gene Cassettes among Salmonella Strains Isolated from Imported Seafood

2008 ◽  
Vol 75 (4) ◽  
pp. 1192-1196 ◽  
Author(s):  
Ashraf A. Khan ◽  
Elizabeth Ponce ◽  
M. S. Nawaz ◽  
Chorng-Ming Cheng ◽  
Junaid A. Khan ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Southern Hybridization ◽  
Multidrug Resistant ◽  
The Philippines ◽  
Streptomycin Resistance ◽  
Class 1 Integron ◽  
Class 1 ◽  
Identification And Characterization ◽  
Multiple Antibiotics

ABSTRACT A total of 210 Salmonella isolates, representing 64 different serovars, were isolated from imported seafood samples, and 55/210 isolates were found to be resistant to at least one antibiotic. Class 1 integrons from three multidrug-resistant Salmonella enterica strains (Salmonella enterica serovars Newport [strain 62], Typhimurium var. Copenhagen [strain 629], and Lansing [strain 803], originating from Hong Kong, the Philippines, and Taiwan, respectively) were characterized. Southern hybridization of plasmids isolated from these strains, using a class 1 integron probe, showed that trimethoprim-sulfamethoxazole and streptomycin resistance genes were located on a megaplasmid in strain 629. Our study indicates that imported seafood could be a reservoir for Salmonella isolates resistant to multiple antibiotics.

scholarly journals Characterization of multidrug-resistant and extended-spectrum β-lactamase-producing Klebsiella pneumoniae strains from Malaysian hospitals

2009 ◽  
Vol 58 (11) ◽  
pp. 1463-1469 ◽  
Author(s):  
King Ting Lim ◽  
Chew Chieng Yeo ◽  
Rohani Md Yasin ◽  
Ganeswrie Balan ◽  
Kwai Lin Thong
Keyword(s):  
Klebsiella Pneumoniae ◽  
Dna Fingerprinting ◽  
Antibiotic Resistance Gene ◽  
Multidrug Resistant ◽  
Pcr Detection ◽  
Management Problem ◽  
Class 1 Integron ◽  
Extended Spectrum ◽  
Class 1 ◽  
Encoding Genes

The emergence of multidrug-resistant (MDR) and extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae poses a serious antibiotic management problem as resistance genes are easily transferred from one organism to another. Fifty-one strains of K. pneumoniae isolated from sporadic cases in various hospitals throughout Malaysia were analysed by antimicrobial susceptibility testing, PCR detection of ESBL-encoding genes and DNA fingerprinting. Although 27 of the 51 K. pneumoniae strains were MDR (i.e. resistant to three or more classes of antibiotics), the majority of the strains (98 %) were sensitive to imipenem. PCR detection using ESBL gene-specific primers showed that 46 of the K. pneumoniae strains harboured bla SHV, 19 harboured bla CTX-M, 5 harboured bla OXA-1 and 4 harboured bla TEM-1. Class 1 integron-encoded intI1 integrase was detected in 21 of the 51 K. pneumoniae strains and amplification of the integron 5′CS region showed the presence of several known antibiotic resistance gene cassettes of various sizes. Results of conjugation and transformation experiments indicated that some of the ESBL-encoding genes (i.e. bla SHV, bla CTX-M and bla TEM-1) were transmissible and were likely plasmid-encoded. DNA fingerprinting using PFGE and PCR-based methods indicated that the 51 K. pneumoniae strains were genetically diverse and heterogeneous.

scholarly journals Genetic characterization of Shigella spp. isolated from diarrhoeal and asymptomatic children

2014 ◽  
Vol 63 (7) ◽  
pp. 903-910 ◽  
Author(s):  
Santanu Ghosh ◽  
Gururaja P. Pazhani ◽  
Swapan Kumar Niyogi ◽  
James P. Nataro ◽  
Thandavarayan Ramamurthy
Keyword(s):  
Virulence Genes ◽  
Multidrug Resistant ◽  
Class 1 Integron ◽  
Genetic Characteristics ◽  
Asymptomatic Children ◽  
Class 1 ◽  
Shigella Spp ◽  
Encoding Gene ◽  
Encoding Genes

Phenotypic and genetic characteristics of Shigella spp. isolated from diarrhoeal and asymptomatic children aged up to 5 years were analysed in this study. In total, 91 and 17 isolates were identified from diarrhoeal (case) and asymptomatic (control) children, respectively. All the isolates were tested for antimicrobial resistance, the presence of integrons, plasmid-mediated quinolone resistance (PMQR), virulence-associated genes and Shigella pathogenicity island (SH-PAI). The majority of the Shigella spp. from cases (68.1 %) and controls (82.3 %) were found to be resistant to fluoroquinolones. Integron carriage was detected more in cases (76.9 %) than in controls (35.5 %). Atypical class 1 integron was detected exclusively in Shigella flexneri from cases but not from the controls. PMQR genes such as aac(6′)-Ib-cr and qnrS1 were detected in 82.4  and 14.3 % of the isolates from cases and in 53 and 17.6 % in controls, respectively. Shigella isolates from cases as well as from controls were positive for the invasive plasmid antigen H-encoding gene ipaH. The other virulence genes such as virF, sat, setA, setB, sen and ial were detected in Shigella isolates in 80.2, 49.4, 27.4, 27.4, 80.2 and 79.1 % of cases and in 64.7, 52.9, 17.6, 17.6, 64.7 and 64.7 % of controls, respectively. The entire SH-PAI was detected in S. flexneri serotype 2a from cases and controls. In an isolate from a control child, the SH-PAI was truncated. Integrons, PMQR and virulence-encoding genes were detected more frequently in cases than in controls. In diarrhoea endemic areas, asymptomatic carriers may play a crucial role in the transmission of multidrug-resistant Shigella spp. with all the putative virulence genes.

scholarly journals Class 1 integron element in Thai Acinetobacter baumannii reveals a linkage to the European clone I

2012 ◽  
Vol 1 (1) ◽  
pp. 24-28
Author(s):  
B Thapa ◽  
C Tribuddharat ◽  
S Srifuengfung ◽  
C Dhiraputra
Keyword(s):  
Acinetobacter Baumannii ◽  
Blot Hybridization ◽  
Variable Region ◽  
Southern Blot Hybridization ◽  
Multidrug Resistant ◽  
Dot Blot ◽  
Class 1 Integron ◽  
Integrase Gene ◽  
Class 1 ◽  
Resistance Island

BACKGROUND: Class 1 integron element is innate to most of the multidrug resistant Acinetobacter baumannii and its spread is common among international clones worldwide. The aim of this study was to document the presence of blaVEB-1 harboring class 1 integron element and its gene cassettes in Thai A. baumannii in relation to A. baumannii European clone I, AYE strain. MATERIALS AND METHODS: Thirty seven carbapenem resistant A. baumannii isolates identified in routine microbiology laboratory of Siriraj Hospital, Bangkok were studied. The dot blot hybridization was performed to detect class 1 integron element integrase gene. PCR was used to amplify blaVEB-1, arr2, cmlA, blaOXA-10 resistance cassettes, and variable region of class 1 integron element. blaVEB-1 gene was localized by southern blot hybridization. RESULTS: The prevalence of class 1 integron element was 86.48% in the isolates studied. The blaVEB-1 was present in 7 isolates however the location of blaVEB-1 gene was different in different isolate. Four isolates (Ab03-168, Ab04-28, Ab08-20, and Ab08-22) harbored calss 1 integron element variable region sized 5.5 kb as described in strain AYE. However, blaVEB-1 was only amplified from Ab03-168. The cassette organization in this isolate was 5’CS-aadB-blaVEB-1-arr2-cmlA-blaOXA- 10-aadA1-3’CS. The class 1 integron element similar to the element identified in genomic resistance island, AbaRI of European clone I, AYE was identified in Thai A. baumannii. CONCLUSIONS: blaVEB-1 harboring class 1 integron element with minor cassette variation was identified in Thai A. baumanni isolate which might suggest the spread of this resistant cassette or the spread of the European clone I in Thailand. Monitoring of the global spread of multi-resistant A. baumannii is mandatory to control the spread of resistant genes and this multi-resistant pathogen. DOI: http://dx.doi.org/10.3126/ijim.v1i1.6715Int J Infect Microbiol 2012;1(1):24-28

scholarly journals Complete Genome Sequencing of Acinetobacter baumannii Strain K50 Discloses the Large Conjugative Plasmid pK50a Encoding Carbapenemase OXA-23 and Extended-Spectrum β-Lactamase GES-11

2018 ◽  
Vol 62 (5) ◽  
Author(s):  
Daniel Wibberg ◽  
Ileana P. Salto ◽  
Felix G. Eikmeyer ◽  
Irena Maus ◽  
Anika Winkler ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Genome Sequencing ◽  
Antibiotic Resistance Gene ◽  
Insertion Site ◽  
Multidrug Resistant ◽  
Conjugative Plasmid ◽  
Class 1 Integron ◽  
Content Type ◽  
Carbapenemase Gene ◽  
Class 1

ABSTRACT Multidrug-resistant (MDR) Acinetobacter baumannii strains appeared as serious emerging nosocomial pathogens in clinical environments and especially in intensive care units (ICUs). A. baumannii strain K50, recovered from a hospitalized patient in Kuwait, exhibited resistance to carbapenems and additionally to ciprofloxacin, chloramphenicol, sulfonamides, amikacin, and gentamicin. Genome sequencing revealed that the strain possesses two plasmids, pK50a (79.6 kb) and pK50b (9.5 kb), and a 3.75-Mb chromosome. A. baumannii K50 exhibits an average nucleotide identity (ANI) of 99.98% to the previously reported Iraqi clinical isolate AA-014, even though the latter strain lacked plasmid pK50a. Strain K50 belongs to sequence type 158 (ST158) (Pasteur scheme) and ST499 (Oxford scheme). Plasmid pK50a is a member of the Aci6 (replication group 6 [RG6]) group of Acinetobacter plasmids and carries a conjugative transfer module and two antibiotic resistance gene regions. The transposon Tn 2008 carries the carbapenemase gene bla OXA-23 , whereas a class 1 integron harbors the resistance genes bla GES-11 , aacA4 , dfrA7 , qacE Δ 1 , and sul1 , conferring resistance to all β-lactams and reduced susceptibility to carbapenems and resistance to aminoglycosides, trimethoprim, quaternary ammonium compounds, and sulfamethoxazole, respectively. The class 1 integron is flanked by MITEs (miniature inverted-repeat transposable elements) delimiting the element at its insertion site.

Characterization of carbapenemase-producing Serratia marcescens and whole-genome sequencing for plasmid typing in a hospital in Madrid, Spain (2016–18)

2020 ◽  
Vol 76 (1) ◽  
pp. 110-116
Author(s):  
Blanca Pérez-Viso ◽  
Marta Hernández-García ◽  
Manuel Ponce-Alonso ◽  
María Isabel Morosini ◽  
Patricia Ruiz-Garbajosa ◽  
...  
Keyword(s):  
Serratia Marcescens ◽  
Antibiotic Resistance Gene ◽  
Epidemiological Surveillance ◽  
Class 1 Integron ◽  
Phenotypic Resistance ◽  
Genetic Lineages ◽  
Clonal Relatedness ◽  
Class 1 ◽  
Almost All

Abstract Objectives Carbapenemase-producing Enterobacterales (CPE) are increasingly recognized in nosocomial infections, also affecting ICU patients. We aimed to characterize the carbapenemase-producing Serratia marcescens (CPSm) isolates recovered in our hospital in Madrid (Spain) between March 2016 and December 2018. Methods Overall, 50 isolates from clinical and epidemiological surveillance samples were recovered from 24 patients admitted to the medical ICU and 10 non-ICU-related patients based on their phenotypic resistance. Carbapenemase characterization, antibiotic susceptibility, PFGE clonal relatedness, plasmid characterization, WGS (Illumina-NovaSeq 6000) and phylogenetic analysis were performed. Results A single isolate was finally considered for each patient, except for Patient 8 that was colonized by two different isolates (n = 35). Isolates were characterized as VIM-1 (n = 29) or OXA-48 producers (n = 6). Up to seven genetic lineages were found by PFGE, with dominance of two clones. Plasmid characterization confirmed that almost all CPSm carried the same ∼60 kb IncL OXA-48- or VIM-1-encoding plasmid, which was related to the globally disseminated IncL-pOXA-48a. WGS allowed plasmid reconstruction with two variants: IncL-pVIM-1 (∼65 kb) and IncL-pOXA-48 (∼62 kb). blaOXA-48–Tn1999 (∼5 kb) was the unique antibiotic resistance gene in pOXA-48, whereas pVIM-1 plasmids (∼8 kb) harboured a class 1 integron containing 5′-blaVIM-1+aacA4+dfrB1+aadA1+catB2+qacEDelta1+sul1-3′. Conclusions Our results confirm the dissemination of CPSm within our institution in both ICU and non-ICU environments, representing two prevalent CPSm clones, and the same IncL-pOXA-48 plasmid previously described in other Enterobacterales, but containing the blaVIM-1 gene. This also reinforces the relevance of species different from Klebsiella pneumoniae or Escherichia coli in the CPE landscape and circulating lineages and plasmids in local CPE epidemiology.

scholarly journals AAC(6′)-Iaf, a Novel Aminoglycoside 6′-N-Acetyltransferase from Multidrug-Resistant Pseudomonas aeruginosa Clinical Isolates

2009 ◽  
Vol 53 (6) ◽  
pp. 2327-2334 ◽  
Author(s):  
Tomoe Kitao ◽  
Tohru Miyoshi-Akiyama ◽  
Teruo Kirikae
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Decubitus Ulcer ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Upstream Region ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Class 1 ◽  
Layer Chromatography

ABSTRACT We report here the characterization of a novel aminoglycoside resistance gene, aac(6′)-Iaf, present in two multidrug-resistant (MDR) Pseudomonas aeruginosa clinical isolates. These isolates, IMCJ798 and IMCJ799, were independently obtained from two patients, one with a urinary tract infection and the other with a decubitus ulcer, in a hospital located in the western part of Japan. Although the antibiotic resistance profiles of IMCJ798 and IMCJ799 were similar to that of MDR P. aeruginosa IMCJ2.S1, which caused outbreaks in the eastern part of Japan, the pulsed-field gel electrophoresis patterns for these isolates were different from that for IMCJ2.S1. Both IMCJ798 and IMCJ799 were found to contain a novel chromosomal class 1 integron, In123, which included aac(6′)-Iaf as the first cassette gene. The encoded protein, AAC(6′)-Iaf, was found to consist of 183 amino acids, with 91 and 87% identity to AAC(6′)-Iq and AAC(6′)-Im, respectively. IMCJ798, IMCJ799, and Escherichia coli transformants carrying a plasmid containing the aac(6′)-Iaf gene and its upstream region were highly resistant to amikacin, dibekacin, and kanamycin but not to gentamicin. The production of AAC(6′)-Iaf in these strains was confirmed by Western blot analysis. Thin-layer chromatography indicated that AAC(6′)-Iaf is a functional acetyltransferase that specifically modifies the amino groups at the 6′ positions of aminoglycosides. Collectively, these findings indicate that AAC(6′)-Iaf contributes to aminoglycoside resistance.

scholarly journals Characterization of Multidrug Resistant ESBL-ProducingEscherichia coliIsolates from Hospitals in Malaysia

2009 ◽  
Vol 2009 ◽  
pp. 1-10 ◽  
Author(s):  
King-Ting Lim ◽  
Rohani Yasin ◽  
Chew-Chieng Yeo ◽  
Savithri Puthucheary ◽  
Kwai-Lin Thong
Keyword(s):  
Antibiotic Resistance ◽  
Random Amplified Polymorphic Dna ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Public Hospitals ◽  
Class 1 Integron ◽  
E Coli ◽  
Antibiotic Management ◽  
Class 1

The emergence ofEscherichia colithat produce extended spectrumβ-lactamases (ESBLs) and are multidrug resistant (MDR) poses antibiotic management problems. Forty-sevenE. coliisolates from various public hospitals in Malaysia were studied. All isolates were sensitive to imipenem whereas 36 were MDR (resistant to 2 or more classes of antibiotics). PCR detection using gene-specific primers showed that 87.5% of the ESBL-producingE. coliharbored theblaTEMgene. Other ESBL-encoding genes detected wereblaOXA,blaSHV, andblaCTX-M. Integron-encoded integrases were detected in 55.3% of isolates, with class 1 integron-encodedintI1integrase being the majority. Amplification and sequence analysis of the5′CS region of the integrons showed known antibiotic resistance-encoding gene cassettes of various sizes that were inserted within the respective integrons. Conjugation and transformation experiments indicated that some of the antibiotic resistance genes were likely plasmid-encoded and transmissible. All 47 isolates were subtyped by PFGE and PCR-based fingerprinting using random amplified polymorphic DNA (RAPD), repetitive extragenic palindromes (REPs), and enterobacterial repetitive intergenic consensus (ERIC). These isolates were very diverse and heterogeneous. PFGE, ERIC, and REP-PCR methods were more discriminative than RAPD in subtyping theE. coliisolates.

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