INCF plasmids responsible by dissemination of blaKPC gene among enterobacteriaceae (original) (raw)
Related papers
Frontiers in Microbiology
Nosocomial infections caused by multidrug-resistant (MDR) Klebsiella pneumoniae are a major health problem worldwide. The aim of this study was to describe NDM-1-producing K. pneumoniae strains causing bacteremia in a tertiary referral hospital in Mexico. MDR K. pneumoniae isolates were screened by polymerase chain reaction for the presence of resistance genes. In resistant isolates, plasmids were identified and conjugation assays were performed. Clonal diversity and the sequence types were determined by pulsed-field gel electrophoresis and multilocus sequence typing. A total of 80 K. pneumoniae isolates were collected from patients with bacteremia over a 1-year period. These isolates showed a level of resistance of 59% (47/80) to aztreonam, 56–60% (45–48/80) to cephalosporins, 54% (43/80) to colistin and 12.5% (10/80) to carbapenems. The carbapenem resistant isolates were blaNDM–1 carriers and negative for blaKPC, blaNDM, blaIMP, blaVIM and blaOXA–48–like carbapenemases genes. Conj...
International Journal of Infectious Diseases, 2016
New Delhi metallo-b-lactamase (NDM)-producing Klebsiella pneumoniae isolates have spread globally, causing infections with a significant and high mortality rate. 1 Over a period of 15 months, from September 2013 to December 2014, a total of 157 Klebsiella spp isolates were recovered from different clinical specimens processed at the microbiology laboratory of Theodor Bilharz Research Institute (TBRI), a tertiary care hospital in Egypt. Thirteen of them were found to be resistant to imipenem or meropenem by disk diffusion and Vitek2 system (bioMé rieux, Marcy L'Etoile, France). Eight of these isolates were available for this study. Minimum inhibitory concentrations (MICs) of a set of antibiotics were determined by standardized broth microdilution method following the guidelines of the Clinical and Laboratory Standards Institute (CLSI). 2 Those of ertapenem and meropenem were also performed by Etest strip method (bioMé rieux, Marcy L'Etoile, France) (Table 1). Data were interpreted according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, 3 except for nalidixic acid, which was categorized according to CLSI standards. 2 One isolate (K4) was susceptible to gentamicin, ciprofloxacin, and nalidixic acid (Table 1). The remaining seven isolates were resistant to cephalosporins, carbapenems, quinolones, and aminoglycosides. All isolates retained susceptibility to both colistin and tigecycline. Discrepancies between the broth microdilution and Etest results were noted regarding carbapenems (Table 1). Such discordance between the two methods has been reported previously in VIM-1producing K. pneumoniae. 4 PCR was performed, followed by sequencing for the genes coding for extended-spectrum b-lactamases (bla SHV , bla TEM , bla CTX-M), plasmid-mediated AmpC b-lactamases, carbapenemases (bla KPC , bla IMP , bla VIM , bla NDM , bla OXA-48), plasmid-mediated quinolone resistance (qnrA, qnrB, qnrS), aminoglycoside-modifying enzymes (aac(3)-Ia, aac(3)-IIa, aac(3)-IVa, aac(6 0)-Ib ant(2 00)-Ia, aph(3 0)-Ia, aph(3 0)-IIa, aph-(3 0)-VIa), and methyltransferases (npmA, armA, rmtB, rmtC, and rmtF). 5-8
Revista da Sociedade Brasileira de Medicina Tropical
Introduction: Antibiotic resistance in carbapenemase-producing Klebsiella pneumoniae is acquired and disseminated mainly by plasmids. Therefore, we aimed to investigate the occurrence of carbapenemase genes, analyze the genetic diversity by ERIC-PCR, and examine the most common plasmid incompatibility groups (Incs) in clinical isolates of K. pneumoniae from colonization and infection in patients from a hospital in Brazil. Methods: Twenty-seven isolates of carbapenem-resistant K. pneumoniae were selected and screened for the presence of carbapenemase genes and Incs by PCR, followed by amplicon sequencing. Results: The bla KPC and bla NDM genes were detected in 24 (88.8 %) and 16 (59.2 %) of the isolates, respectively. Thirteen isolates (48.1 %) were positive for both genes. The IncFIB (92.6 %) and IncQ (88.8 %) were the most frequent plasmids, followed by IncA/C, IncHI1B, and IncL/M, indicating that plasmid variability existed in these isolates. To our knowledge, this is the first report of IncHI1B in Brazil. We found eight isolates with clonal relationship distributed in different sectors of the hospital. Conclusions: The accumulation of resistance determinants, the variability of plasmid Incs, and the clonal dissemination detected in K. pneumoniae isolates demonstrate their potential for infection, colonization, and the dissemination of different resistance genes and plasmids.
The role of the bla KPC gene in antimicrobial resistance of Klebsiella pneumoniae
Iranian Journal of Microbiology, 2019
Background and Objectives: Klebsiella pneumoniae isolates that produce K. pneumoniae carbapenemase (KPC) have be- come a grave concern for the treatment of infections. KPC-producing strains are not only able to hydrolyze carbapenems but are also resistant to a variety of β-lactam and non-β-lactam antibiotics. The present study evaluated the prevalence of bla in K. pneumoniae infections and determined the antimicrobial susceptibility of the isolates. Materials and Methods: The K. pneumoniae isolates were identified by biochemical tests and confirmed by genotyping. The modified Hodge test (MHT) was performed to detect carbapenemases, and antimicrobial susceptibility was determined for all isolates by the disc diffusion method. Also, for MHT-positive isolates, supposed to carbapenemases isolates, broth microdilution method was used to measure the minimum inhibitory concentrations (MICs) of meropenem and colistin. Results: The bla KPC genotypic evaluation revealed that only 5 of 96 isol...
Journal of Antimicrobial Chemotherapy, 2010
We describe 12 VIM-1-producing strains (7 Enterobacter cloacae, 2 Klebsiella pneumoniae and 3 clonal Klebsiella oxytoca strains) detected among clinically relevant Enterobacteriaceae isolates from routine cultures at the Hospital del Mar (Barcelona, Spain) from December 2006 to May 2007. Methods: Susceptibility to carbapenems was evaluated with the MicroScan system. b-Lactamases were identified by PCR and sequencing. Clonal relationships between the isolates were analysed by PFGE. Transferability of the enzymes was tested by conjugation. Plasmid characterization was performed by PCR-based replicon typing and PFGE with S1 nuclease digestion of whole genomic DNA. The PFGE gels were then transferred and hybridized. Results: The disc diffusion method correctly identified five of the seven E. cloacae isolates as intermediate or resistant strains. All isolates produced the VIM-1 enzyme. Three E. cloacae and three K. oxytoca strains were also CTX-M-9-producing strains, and one E. cloacae was also a CTX-M-3-producing strain. The plasmids carrying the bla VIM gene, of unknown incompatibility group, had a size of 75 kb (eight strains) or 40 kb (three strains) and also contained the qnrS and the aac(6 0)-Ib-cr genes. In the remaining strain the bla VIM-1 gene was found in an HI2 plasmid of 290 kb together with bla CTX-M-9 , qnrA, qnrS and the aac(6 0)-Ib-cr genes. Conclusions: The results showed a linkage between the bla VIM-1 and the qnrS and the aac(6 0)-Ib-cr genes, and between the bla CTX-M-9 and the qnrA genes.
Background: To assess antimicrobial susceptibility of extended-spectrum β-lactamase-(ESBL-) producing Klebsiella pneumoniae and Escherichia coli isolates from Hospital Tengku Ampuan Afzan (HTAA), as well as to identify ESBL genes. Methods: Non-duplicate K. pneumoniae and E. coli isolates were recovered from various clinical samples. Isolates were screened for antimicrobial resistance by disc diffusion method. Isolates resistant to oxyimino-cephalosporins were subjected to phenotypic ESBL production. Detection of resistance genes was then performed using primers specific for ESBL genes (blaCTX-M, blaSHV and blaTEM). Results: Piperacillin/tazobactam and carbapenems remained the active β-lactam antibiotic against K. pneumoniae and E. coli. ESBLs were detected among 35.5% (39/110) of K. pneumoniae and 18.8% (28/149) of E. coli isolates. CTX-M β-lactamase was detected in 90% of all ESBL-positive isolates, whereas blaSHV and blaTEM genes were found among 56% and 52% of them, respectively. Twenty-eight percent (28%) of the total ESBL-positive isolates harboured the three ESBL genes, while 50% carried two of the tested ESBL genes. Conclusion: ESBLs encoded by at least one ESBL genes are frequently isolated among K. pneumoniae and E. coli in HTAA. The significant proportion rate of these resistant determinants is alarming, thus monitoring their transmission and dissemination is essential to control it at an early phase.
Revista da Sociedade Brasileira de Medicina Tropical
Introduction: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. Methods: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. Results: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTE KPC-IId). It was detected on a plasmid belonging to the IncQ1 group. Conclusions: To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTE KPC-IId element carried by plasmid IncQ1 from infections in Brazil.
Antimicrobial Agents and Chemotherapy, 2014
We sequenced the oldest bla KPC-2 -bearing plasmid isolated in Brazil and another plasmid also carried by a Klebsiella pneumoniae strain of sequence type 442 (ST442), isolated 52 months later. Both plasmids present an IncN backbone and few acquired regions. Because the 2005 plasmid presented deletions and a truncated gene within Tn4401b compared to the 2009 plasmid, we can thus infer that IncN bla KPC-2 -bearing plasmids pFCF1305 and pFCF3SP had a common ancestor circulating in Brazil prior to May 2005. E ven though the first description of KPC-producing Klebsiella pneumoniae in Brazil was published in 2009, describing a strain isolated from a clinical sample in 2006 (1), a later publication reported the presence of a KPC-bearing strain isolated in May 2005
Antimicrobial Agents and Chemotherapy, 2002
Over a 12-year period (1989 to 2000), 159 Klebsiella pneumoniae isolates harboring extended-spectrum -lactamases (ESBLs) (4.8% of the total number of K. pneumoniae isolates obtained) were recovered from 58 patients, who were mainly hospitalized in intensive care and surgery units. For 62 representative isolates from 58 patients, 31 clonal types harboring TEM-4 (n ؍ 5), SHV-2 (n ؍ 7), SHV2a (n ؍ 4), SHV-5 (n ؍ 1), CTX-M-10 (n ؍ 13), or CTX-M-9 (n ؍ 1) -lactamases were identified by pulsed-field gel electrophoresis. This is the first report to document the presence of the CTX-M-10 or the CTX-M-9 -lactamase in K. pneumoniae. These -lactamases were previously identified in Escherichia coli isolates from Spain. Only two of five K. pneumoniae TEM-4 clones caused more than a single case of infection, with one of them spreading for 9 months. A single plasmid was detected among these TEM-4 clones. Only two of seven K. pneumoniae clones containing SHV-2 and three of four strains harboring SHV-2a were detected in more than one case of infection. Plasmids encoding SHV-2 or SHV-2a were unrelated. Four of 13 K. pneumoniae CTX-M-10 clones were found in more than one patient, with two of them recovered 2 and 5 years apart. As in the case of the SHV-2 isolates, we were unable to document a common transmissible genetic element that could explain the polyclonal structure of our isolates. Nevertheless, the spread of a single gene may be suggested by the presence of a conserved set of noncoding polymorphisms in the sequences. Most ESBL-producing K. pneumoniae clones were ephemeral, being poorly selected and maintained in the hospital setting, but the genes encoding ESBL persisted successfully over the years that the strains were recovered, probably as a minority gene population in the hospital metagenome.
Antibiotics
The emergence of blaKPC-2 and blaNDM-1 producing Klebsiella pneumoniae represents a great problem in many Egyptian hospitals. One hundred and twenty-six K. pneumoniae isolates from patients admitted to Assiut University Hospital were identified by an API20E kit. Carbapenemase-producing K. pneumoniae (CPKP) was detected by the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method (eCIM), and an E-test. Based on the polymerase chain reaction, all isolates were negative for bla-VIM-1 and bla-IMP-1, fifteen of these isolates were positive for both blaKPC-2 and blaNDM-1, two isolates were positive for blaKPC-2 only, and twenty-eight isolates were positive for bla-NDM-1 only. Although one isolate was positive for the string test, all CPKP isolates were negative for capsular genes. Only 71.1% of CPKP transferred their plasmids to their corresponding transconjugants (E. coli J53). The resistance patterns of the clinical isolates and their transconj...