scholarly journals Molecular Identification of Aminoglycoside-Modifying Enzymes and Plasmid-Mediated Quinolone Resistance Genes among Klebsiella pneumoniae Clinical Isolates Recovered from Egyptian Patients

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Mohamed F. El-Badawy ◽  
Wael M. Tawakol ◽  
Shaymaa W. El-Far ◽  
Ibrahim A. Maghrabi ◽  
Saleh A. Al-Ghamdi ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Quinolone Resistance ◽  
Clinical Isolates ◽  
Phylogenetic Groups ◽  
Aminoglycoside Resistance ◽  
Inappropriate Use ◽  
Genes Encoding

Inappropriate use of antibiotics in clinical settings is thought to have led to the global emergence and spread of multidrug-resistant pathogens. The goal of this study was to investigate the prevalence of genes encoding aminoglycoside resistance and plasmid-mediated quinolone resistance among clinical isolates of Klebsiella pneumoniae. All K. pneumoniae isolates were phenotypically identified using API 20E and then confirmed genotypically through amplification of the specific K. pneumoniae phoE gene. All isolates were genotyped by the enterobacterial repetitive intergenic consensus polymerase chain reaction technique (ERIC-PCR). Antibiotic susceptibility testing was done by a modified Kirby-Bauer method and broth microdilution. All resistant or intermediate-resistant isolates to either gentamicin or amikacin were screened for 7 different genes encoding aminoglycoside-modifying enzymes (AMEs). In addition, all resistant or intermediate-resistant isolates to either ciprofloxacin or levofloxacin were screened for 5 genes encoding the quinolone resistance protein (Qnr), 1 gene encoding quinolone-modifying enzyme, and 3 genes encoding quinolone efflux pumps. Biotyping using API 20E revealed 13 different biotypes. Genotyping demonstrated that all isolates were related to 2 main phylogenetic groups. Susceptibility testing revealed that carbapenems and tigecycline were the most effective agents. Investigation of genes encoding AMEs revealed that acc(6′)-Ib was the most prevalent, followed by acc(3′)-II, aph(3′)-IV, and ant(3′′)-I. Examination of genes encoding Qnr proteins demonstrated that qnrB was the most prevalent, followed by qnrS, qnrD, and qnrC. It was found that 61%, 26%, and 12% of quinolone-resistant K. pneumoniae isolates harbored acc(6′)-Ib-cr, oqxAB, and qebA, respectively. The current study demonstrated a high prevalence of aminoglycoside and quinolone resistance genes among clinical isolates of K. pneumoniae.

scholarly journals The Molecular Epidemiology of Resistance to Antibiotics among Klebsiella pneumoniae Isolates in Azerbaijan, Iran

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Mehdi Kashefieh ◽  
Hassan Hosainzadegan ◽  
Shabnam Baghbanijavid ◽  
Reza Ghotaslou
Keyword(s):  
Drug Resistance ◽  
Klebsiella Pneumoniae ◽  
Molecular Epidemiology ◽  
Resistance Genes ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Aminoglycoside Resistance ◽  
State Hospitals ◽  
Aminoglycoside Resistance Genes ◽  
Hospital Acquired

Introduction. Klebsiella pneumoniae (K. pneumoniae) is one of the leading causes of hospital-acquired and community-acquired infections in the world. This study was conducted to investigate the molecular epidemiology of drug resistance in clinical isolates of K. pneumoniae in Azerbaijan, Iran. Materials and Methods. A total of 100 nonduplicated isolates were obtained from the different wards of Azerbaijan state hospitals, Iran, from 2019 to 2020. Antibiotic susceptibility testing was done. The DNA was extracted, and the PCR for evaluation of the resistance genes was carried out. Results. The highest antibiotic resistance was shown to ampicillin (96%), and the highest susceptibility was shown to tigecycline (9%), and 85% of isolates were multidrug resistant. The most frequent ESBL gene in the tested isolates was blaSHV-1 in 58%, followed by blaCTXM-15 (55%) and blaSHV-11(42%). The qepA, oqxB, and oqxA genes were found to be 95%, 87.5%, and 70%, respectively. We detected tetB in 42%, tetA in 32%, tetD in 21%, and tetC in 16%. Seventy isolates were resistant to co-trimoxazole, and the rate of resistance genes was sul1 in 71%, followed by sul2 (43%), dfr (29%), and sul3 (7%). The most common aminoglycoside resistance genes were ant3Ia, aac6Ib, aph3Ib, and APHs in 44%, 32%, 32%, and 31.4%, respectively. The most frequent resistance gene to fosfomycin was fosA (40%) and fosX (40%) followed by fosC (20%). Conclusion. The results of this study indicate the high frequency of drug resistance among K. pneumoniae isolated from hospitals of Azerbaijan state. The present study shows the presence of high levels of drug-resistant genes in various antibiotics, which are usually used in the treatment of infections due to K. pneumoniae.

scholarly journals Coexistence of aminoglycoside resistance genes in CTX-M-producing isolates of Klebsiella pneumoniae in Bushehr province, Iran

2021 ◽  
Author(s):  
Behrouz Latifi ◽  
Saeed Tajbakhsh ◽  
Leila Ahadi ◽  
Forough Yousefi
Keyword(s):  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Confirmatory Test ◽  
M Gene ◽  
Aminoglycoside Resistance ◽  
Genetic Groups ◽  
Genes Encoding ◽  
Resistance Patterns ◽  
Aminoglycoside Resistance Genes ◽  
Antibiotic Resistance Patterns

Background and Objectives: Increasing the rate of extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae has given rise to a major healthcare issue in clinical settings over the past few years. Treatment of these strains is hardly effective since the plasmid encoding ESBL may also carry other resistance genes including aminoglycosides. The current study aimed to evaluate the prevalence of ESBL-producing K. pneumoniae and investigate the coexistence of Cefoxitamase-Munich (bla ) with aminoglycoside-modifying enzyme (AME) genes, aac(3)IIa as well as aac(6′)Ib, in CTX‑M‑producing K. pneumoniae isolated from patients in Bushehr province, Iran. Materials and Methods: A total of 212 K. pneumoniae isolates were collected and confirmed using polymerase chain re‑ action (PCR) of the malate dehydrogenase gene. Isolates were screened for production of ESBL. Phenotypic confirmatory test was performed using combined disk test. The genes encoding CTX-M groups and AME genes, aac(3)IIa and aac(6′)Ib, were investigated by PCR. Results: The ESBL phenotype was detected in 56 (26.4%) K. pneumoniae isolates. Moreover, 83.9% of ESBL-producing isolates carried the genes for CTX-M type β-lactamases, which were distributed into the two genetic groups of CTX-M-1 (97.8%)- and CTX-M-2 (2.1%)-related enzymes. Notably, among K. pneumoniae isolates containing the blaCTX‑M gene, 68.08% of isolates harbored AME genes. In addition, the coexistence of bla in 46.8% of CTX-M-producing K. pneumoniae isolates. Conclusion: This study provides evidence of a high prevalence of AME genes in CTX-M- producing K. pneumoniae iso‑ lates; therefore, in the initial empirical treatment of infections caused by ESBL-KP in regions with such antibiotic resistance patterns, aminoglycoside combination therapy should be undertaken carefully.

scholarly journals First Report of an OXA-48-Producing Multidrug-Resistant Proteus mirabilis Strain from Gaza, Palestine

2015 ◽  
Vol 59 (7) ◽  
pp. 4305-4307 ◽  
Author(s):  
Liang Chen ◽  
Nahed Al Laham ◽  
Kalyan D. Chavda ◽  
Jose R. Mediavilla ◽  
Michael R. Jacobs ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Proteus Mirabilis ◽  
Draft Genome ◽  
Multidrug Resistant ◽  
Aminoglycoside Resistance ◽  
Content Type ◽  
Antimicrobial Resistance Genes ◽  
Carbapenemase Gene ◽  
Genes Encoding ◽  
Draft Genome Sequencing

ABSTRACTWe report the first multidrug-resistantProteus mirabilisstrain producing the carbapenemase OXA-48 (Pm-OXA-48) isolated at Al-Shifa hospital in Gaza, Palestine. Draft genome sequencing of Pm-OXA-48 identified 16 antimicrobial resistance genes, encoding resistance to β-lactams, aminoglycosides, fluoroquinolones, phenicols, streptothricin, tetracycline, and trimethoprim-sulfamethoxazole. Complete sequencing of theblaOXA-48-harboring plasmid revealed that it is a 72 kb long IncL/M plasmid, harboring carbapenemase geneblaOXA-48, extended spectrum β-lactamase geneblaCTX-M-14, and aminoglycoside resistance genesstrA,strB, andaph(3′)-VIb.

Prevalence of Genes Encoding Aminoglycoside-Modifying Enzymes and armA among Acinetobacter baumannii Clinical Isolates in Alexandria, Egypt

2021 ◽  
Vol 21 ◽  
Author(s):  
Amel ELsheredy ◽  
Zainab Yousif ◽  
Ebtesam Elghazzawi ◽  
Ahmed Elmenshawy ◽  
Abeer Ghazal
Keyword(s):  
Acinetobacter Baumannii ◽  
Resistance Genes ◽  
Antimicrobial Agents ◽  
Clinical Isolates ◽  
Aminoglycoside Resistance ◽  
High Propensity ◽  
Genes Encoding ◽  
Resistance Patterns ◽  
Aminoglycoside Resistance Genes ◽  
High Level

Introduction: Acinetobacter baumannii (A.baumannii ) is a ubiquitous pathogen responsible for serious infections in hospitalized patients with a high propensity to develop resistance to antimicrobial agents. The study aimed to determine the antimicrobial resistance patterns and the prevalence of aminoglycoside resistance genes among A. baumannii clinical isolates from patients in different intensive care units (ICUs) in Alexandria, Egypt. Methods: A total of 100 A. baumannii isolates collected from ICU patients were confirmed as A. baumannii by VITEK 2 and the presence of the blaOXA-51 gene has been reported. Antimicrobial susceptibility testing was performed and Multiplex PCR was done for the detection of aminoglycoside resistance genes. Results: Most of the isolates (82%) were resistant to all tested aminoglycosides; resistance was higher for kanamycin and neomycin, followed by amikacin. The predominant AMEs were aphA6 and aphA1 in 86% and 67% of the isolates, respectively; aacA4 and aacC1 were detected in 37% and 8%, respectively, while aadA1 and aadB were present in 34% and 4%, respectively. Furthermore, armA gene was detected in 83% of the isolates. Conclusion: The results of this study revealed a high level carriage of armA and AMEs which limit the usage of aminoglycoside as a treatment option for A. baumannii and makes treatment extremely difficult.

scholarly journals Novel Gyrase Mutations in Quinolone-Resistant and -Hypersusceptible Clinical Isolates of Mycobacterium tuberculosis: Functional Analysis of Mutant Enzymes

2006 ◽  
Vol 50 (1) ◽  
pp. 104-112 ◽  
Author(s):  
Alexandra Aubry ◽  
Nicolas Veziris ◽  
Emmanuelle Cambau ◽  
Chantal Truffot-Pernot ◽  
Vincent Jarlier ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Dna Cleavage ◽  
Multidrug Resistant ◽  
Quinolone Resistance ◽  
Clinical Isolates ◽  
Resistance Mutations ◽  
Drug Induced ◽  
Mutant Proteins ◽  
Highly Active ◽  
Genes Encoding

ABSTRACT Mutations in the DNA gyrase GyrA2GyrB2 complex are associated with resistance to quinolones in Mycobacterium tuberculosis. As fluoroquinolones are being used increasingly in the treatment of tuberculosis, we characterized several multidrug-resistant clinical isolates of M. tuberculosis carrying mutations in the genes encoding the GyrA or GyrB subunits associated with quinolone resistance or hypersusceptibility. In addition to the reported putative quinolone resistance mutations in GyrA, i.e., A90V, D94G, and D94H, we found that the GyrB N510D mutation was also associated with ofloxacin resistance. Surprisingly, several isolates bearing a novel combination of gyrA T80A and A90G changes were hypersusceptible to ofloxacin. M. tuberculosis GyrA and GyrB subunits (wild type [WT] and mutants) were overexpressed in Escherichia coli, purified to homogeneity, and used to reconstitute highly active gyrase complexes. Mutant proteins were produced similarly from engineered gyrA and gyrB alleles by mutagenesis. MICs, enzyme inhibition, and drug-induced DNA cleavage were determined for moxifloxacin, gatifloxacin, ofloxacin, levofloxacin, and enoxacin. Mutant gyrase complexes bearing GyrA A90V, D94G, and D94H and GyrB N510D were resistant to quinolone inhibition (MICs and 50% inhibitory concentrations [IC50s] at least 3.5-fold higher than the concentrations for the WT), and all, except the GyrB mutant, were less efficiently trapped as a quinolone cleavage complex. In marked contrast, gyrase complexes bearing GyrA T80A or A90G were hypersusceptible to the action of many quinolones, an effect that was reinforced for complexes bearing both mutations (MICs and IC50s up to 14-fold lower than the values for the WT). This is the first detailed enzymatic analysis of hypersusceptibility and resistance in M. tuberculosis.

scholarly journals Genomic evolution of the globally disseminated multidrug-resistant Klebsiella pneumoniae clonal group 147

2022 ◽  
Vol 8 (1) ◽  
Author(s):  
Carla Rodrigues ◽  
Siddhi Desai ◽  
Virginie Passet ◽  
Devarshi Gajjar ◽  
Sylvain Brisse
Keyword(s):  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Antimicrobial Susceptibility ◽  
Type Species ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Antimicrobial Susceptibility Testing ◽  
Content Type ◽  
Genetic Elements ◽  
Link Type

The rapid emergence of multidrug-resistant Klebsiella pneumoniae is being driven largely by the spread of specific clonal groups (CGs). Of these, CG147 includes 7-gene multilocus sequence typing (MLST) sequence types (STs) ST147, ST273 and ST392. CG147 has caused nosocomial outbreaks across the world, but its global population dynamics remain unknown. Here, we report a pandrug-resistant ST147 clinical isolate from India (strain DJ) and define the evolution and global emergence of CG147. Antimicrobial-susceptibility testing following European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines and genome sequencing (Illumina and Oxford Nanopore Technologies, Unicycler assembly) were performed on strain DJ. Additionally, we collated 217 publicly available CG147 genomes [National Center for Biotechnology Information (NCBI), May 2019]. CG147 evolution was inferred within a temporal phylogenetic framework (beast) based on a recombination-free sequence alignment (Roary/Gubbins). Comparative genomic analyses focused on resistance and virulence genes and other genetic elements (BIGSdb, Kleborate, PlasmidFinder, phaster, ICEfinder and CRISPRCasFinder). Strain DJ had a pandrug-resistance phenotype. Its genome comprised the chromosome, seven plasmids and one linear phage-plasmid. Four carbapenemase genes were detected: bla NDM-5 and two copies of bla OXA-181 in the chromosome, and a second copy of bla NDM-5 on an 84 kb IncFII plasmid. CG147 genomes carried a mean of 13 acquired resistance genes or mutations; 63 % carried a carbapenemase gene and 83 % harboured bla CTX-M. All CG147 genomes presented GyrA and ParC mutations and a common subtype I-E CRISPR-Cas system. ST392 and ST273 emerged in 2005 and 1995, respectively. ST147, the most represented phylogenetic branch, was itself divided into two main clades with distinct capsular loci: KL64 (74 %, DJ included, emerged in 1994 and disseminated worldwide, with carbapenemases varying among world regions) and KL10 (20 %, emerged in 2002, predominantly found in Asian countries, associated with carbapenemases NDM and OXA-48-like). Furthermore, subclades within ST147-KL64 differed at the yersiniabactin locus, OmpK35/K36 mutations, plasmid replicons and prophages. The absence of IncF plasmids in some subclades was associated with a possible activity of a CRISPR-Cas system. K. pneumoniae CG147 comprises pandrug-resistant or extensively resistant isolates, and carries multiple and diverse resistance genes and mobile genetic elements, including chromosomal bla NDM-5. Its emergence is being driven by the spread of several phylogenetic clades marked by their own genomic features and specific temporo–spatial dynamics. These findings highlight the need for precision surveillance strategies to limit the spread of particularly concerning CG147 subsets.

scholarly journals First Occurrence of OXA-72-Producing Acinetobacter baumannii in Serbia

2016 ◽  
Vol 60 (10) ◽  
pp. 5724-5730 ◽  
Author(s):  
Laurent Dortet ◽  
Rémy A. Bonnin ◽  
Sandrine Bernabeu ◽  
Lélia Escaut ◽  
Daniel Vittecoq ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Pcr Analysis ◽  
Aminoglycoside Resistance ◽  
Content Type ◽  
A Genome ◽  
16S Rna ◽  
Genes Encoding ◽  
First Occurrence

ABSTRACTHere, we characterized the first OXA-72-producingAcinetobacter baumanniiisolate (designated MAL) recovered from a urine sample from a Serbian patient. Antimicrobial susceptibility testing, plasmid analysis, and whole-genome sequencing (WGS) were performed to fully characterize the resistome of theA. baumanniiMAL clinical isolate. The isolate was multidrug resistant and remained susceptible only to colistin and tigecycline. PCR analysis revealed the presence of the carbapenemase OXA-72, an OXA-40 variant. Extraction by the Kieser method revealed the presence of two plasmids, and one of these, a ca. 10-kb plasmid, harbored theblaOXA-72gene. WGS revealed 206 contigs corresponding to a genome of 3.9 Mbp in size with a G+C content of 38.8%. The isolate belonged to sequence type 492 and to worldwide clone II (WWCII). Naturally occurring β-lactamase-encoding genes (blaADC-25andblaOXA-66) were also identified. Aminoglycoside resistance genes encoding one aminoglycoside adenyltransferase (aadA2), three aminoglycoside phosphatases (strA,strB,aphA6), and one 16S RNA methylase (armA) conferring resistance to all aminoglycosides were identified. Resistance to fluoroquinolones was likely due to mutations ingyrA,parC, andparE. Of note, the resistome matched perfectly with the antibiotic susceptibility testing results.

First description of NDM-1-, KPC-2-, VIM-2- and IMP-4-producing Klebsiella pneumoniae strains in a single Chinese teaching hospital

2014 ◽  
Vol 143 (2) ◽  
pp. 376-384 ◽  
Author(s):  
Y. LIU ◽  
L.-G. WAN ◽  
Q. DENG ◽  
X.-W. CAO ◽  
Y. YU ◽  
...  
Keyword(s):  
16S Rrna ◽  
Klebsiella Pneumoniae ◽  
Teaching Hospital ◽  
Multidrug Resistant ◽  
The Other ◽  
Quinolone Resistance ◽  
Resistance Determinants ◽  
Carbapenem Resistant ◽  
Extended Spectrum ◽  
Chinese Teaching

SUMMARYA total of 180 non-duplicate carbapenem-resistant Klebsiella pneumoniae isolates were recovered from patients hospitalized between December 2010 and January 2012 at a Chinese hospital. Eight KPC-2, four NDM-1, one VIM-2, and five KPC-2 plus IMP-4 producers were identified and all were multidrug resistant due to the presence of other resistance determinants, including extended-spectrum β-lactamases (CTX-M-15, SHV-12), 16S rRNA methylases (armA, rmtB) and plasmid-mediated quinolone-resistance determinants (qnrA, B, S, aac(6′)-Ib-cr). Nine K. pneumoniae clones (Kpn-A1/ST395, Kpn-A3/ST11, Kpn-A2/ST134, Kpn-B/ST263, Kpn-C/ST37, Kpn-D/ST39, Kpn-E/ST1151, Kpn-F/ST890, Kpn-G/ST1153) were identified. blaKPC-2 was located on transferable ~65 kb IncL/M (ST395, ST11, ST134, ST39) and ~100 kb IncA/C (ST37, ST1153, ST890) plasmids, respectively. On the other hand, blaNDM-1 was associated with a ~70 kb IncA/C plasmid (ST263). However, non-typable plasmids of ~40 kb containing blaVIM-2 were detected in the ST1151 clone. This work reports the first co-occurrence of four diverse types of carbapenemase of K. pneumoniae clones from a single hospital in China. IncA/C, IncL/M, and other successful plasmids may be important for the dissemination of carbapenemases, producing a complex epidemiological picture.

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