• BMB Reports
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• v.44 no.4
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• pp.262-266
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• 2011

The aac(6')-Ib gene is the most prevalent gene that encodes aminoglycoside-modifying enzymes and confers resistance to tobramycin, kanamycin, and amikacin. The aac(6')-Ib-cr variant gene can induce resistance against aminoglycoside and fluoroquinolone simultaneously. Two main methods, sequence analysis and the restriction enzyme method, can detect the aac(6')-Ib-cr variant in clinical strains. We collected the 85 strains that were believed to be aac(6')-Ib positive from clinical isolates. Among them, 38 strains were the wild-type; the remaining 47 strains were the aac(6')-Ib-cr variant. Of these 47 strains, 19 simultaneously harbored aac(6')-Ib and aac(6')-Ib-cr. Our study aims to report the characteristics of the 19 strains that simultaneously harbored both genes. This study is the first investigation published in Korea of strains that included both aac(6')-Ib and aac(6')-Ib-cr variant.

• Journal of Life Science
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• v.20 no.6
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• pp.964-967
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• 2010

Plasmid-mediated quinolone resistance (PMQR) determinants have been contributed to quinolone resistance of gram-negative bacteria worldwide. However, little data on the prevalence of these determinants in bacteria from animals are available in Korea. In this study, the prevalence of PMQR genes was investigated with E. coli originating from diseased animals. Among 55 E. coli tested, 11 showed PMQR genes by PCR. The most prevalent genotype was qepA (14.5%), followed by aac(6')-Ib-cr (7.3%) and qnrS (1.8%). Interestingly, two isolates with PMQR genes did not show quinolone resistance in this study. The isolates exhibited higher fluoroquinolone resistance in aac(6')-Ib-cr in combination with qnrS or qepA compared with aac(6')-Ib-cr only. In a conjugal transfer test, PMQR genes were transferred from donor to recipient.

• Korean Journal of Clinical Laboratory Science
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• v.44 no.3
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• pp.155-165
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• 2012

The purpose of this study was to develop a multiplex PCR for the detection of aac(6')-Ib, aph(3')-Ia, and ant(2")-Ia; the genes that encode the most clinically relevant aminoglycoside modifying enzymes (AMEs) in Gram-negative bacteria. Clinical isolates of 80 E. coli and 23 K. pneumoniae from tertiary university hospital were tested by multiplex PCR. The most prevalent AME gene was aac(6')-Ib which was found in 22.3% of the isolates. Of the total 80 E. coli isolates, 1 isolate was found to contain both aph(3')-Ia and ant(2")-Ia simultaneouly. Of the total 23 K. pneumoniae isolates, 2 isolates were found to contain both aac(6')-Ib and aph(3')-Ia, and 1 isolate was found to contain both aac(6')-Ib and ant(2")-Ia simultaneously. Annual (2005~2009) analysis of isolates that contain the AME genes were of no correlation. The sensitivity and specificity of multiplex PCR in detecting AME genes was 94.4% (34 of 36 cases) and 100%, respectively. We suggest the multiplex PCR method we developed could be highly sensitive and specific in detecting the AME genes of E. coli and K. pneumoniae. This study could be the first published investigation in which the multiplex PCR method detects aac(6')-Ib, aph(3')-Ia, and ant(2")-Ia genes.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.2
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• pp.136-142
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• 2020

The antimicrobial resistance rate in bacteria has increased over the last several decades. The transfer of antimicrobial resistant determinants on mobile genetic elements could cause the accelerated emergence and spread of multidrug resistant bacteria. This study investigated the aminoglycoside resistance determinants transferred by mobile genetic elements in a total of 33 aminoglycoside non-susceptible E. coli isolated from clinical specimens in Chungcheong province. 16S ribosomal RNA methyl-transferases (RMTases) and aminoglycoside-modifying enzyme (AME) genes were detected via PCR and DNA sequencing. The most common AME genes were aac(3')-II gene (54.5%), followed by aph(3')-Ia (18.2%) and aac(6')-Ib (15.2%). None of the evaluated RMTase genes were detected in the 33 isolates. Seventeen of the 18 isolates harboring aac(3')-II gene were resistant to gentamicin, and 16 of them were resistant to tobramycin. The 5 isolates harboring aac(6')-Ib gene were all resistant to tobramycin. In this study, we confirmed that one of the important mechanisms of aminoglycoside resistance in E. coli isolated from human is the acquisition of AME genes. Continuing investigations of antimicrobial resistant determinants in bacteria isolated from human may be required to prevent dissemination of antimicrobial resistant bacteria.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1180-1183
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• 2020

The prevalence and characterization of plasmid-mediated quinolone resistance (PMQR) determinants in ciprofloxacin-resistant Escherichia coli isolated from a Korean commercial layer farm were studied. A total of 45 ciprofloxacin-resistant E. coli isolates were recovered and all isolates were multidrug-resistant. Eight isolates have the PMQR genes aac(6')-Ib-cr, qnrS1, and qnrB4, and seven isolates exhibited double amino acid exchange at both gyrA and parC, and have high fluoroquinolone minimum inhibitory concentrations. Five transconjugants demonstrated transferability of PMQR and β-lactamase genes and similar antimicrobial resistance. Because PMQR genes in isolates from commercial layer chickens could enter the food supply and directly affect humans, control of ciprofloxacin resistance is needed.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.3
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• pp.217-224
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• 2021

Fluoroquinolone (FQ) resistant gram-negative pathogens have emerged worldwide, and the recent increase in FQ resistant Escherichia coli is of great concern in Korea. This study investigated FQ resistance determinants and the epidemiological relationship of 56 ciprofloxacin-resistant E. coli isolated from a tertiary hospital in Daejeon, South Korea from June to December 2018. Molecular epidemiology was investigated by multilocus sequence typing (MLST). Polymerase chain reaction (PCR) and sequence analysis were performed to identify chromosomal mutations in the quinolone resistance determining regions (QRDR) of gyrA, gyrB, parC, and parE and to describe the occurrence of the following plasmid-mediated quinolone resistance (PMQR) genes: aac(6)-Ib-cr, qepA, qnrA, qnrB, qnrC, qnrD, and qnrS. MLST analysis showed 12 sequence types (STs) and the most prevalent ST was ST131 (31/56, 55.4%), followed by ST1193 (13/56, 23.2%), and ST405 (3/56, 5.4%). In 56 ciprofloxacin-resistant E. coli isolates, Ser83→Leu and Asp87→Asn in gyrA and Ser80→Ile and Glu84→Val in parC (51.8%, 29/56) were the most frequent amino acid substitutions and aac(6)-Ib-cr (33.9%, 19/56) was the most common PMQR gene. These results of FQ resistance determinants were more frequently observed in ST131 compared with other clones. Continuous monitoring of the epidemiological characteristics of ciprofloxacin-resistant E. coli isolates and further investigation of FQ resistance determinants are necessary.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.210-216
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• 2016

Recently, there has been a considerable increase in the prevalence of CTX-M type extended-spectrum ${\beta}$-lactamase (ESBL)-producing E. coli isolates worldwide, including Korea. To investigate the ESBL genes in the E. coli strains isolated from a university hospital in the Chungcheong area, a study was conducted using PCR amplification and nucleotide sequence analysis of the amplified products to detect the plasmid mediated quinolone resistance (PMQR) genes in ESBL producing E. coli isolates. The number of CTX-M-14 producing isolates was 25 (16.0%) isolates, and of them, 9 (5.8%) isolates also produced CTX-M-15. All CTX-M type ESBL producing E. coli isolates showed resistance to cefotaxime. Twelve (48%) CTX-M type ESBL producing E. coli isolates contained the PMQR genes, 8 contained qnrS1, and 8 contained aac(6')-Ib-cr. Four isolates harbored both qnrS1 and aac(6')-Ib-cr genes. In our study, we confirmed that the plasmid mediated antimicrobial resistant determinants-the ESBL and PMQR genes-were distributed in the E. coli isolates. To prevent further spreading of the resistant genes among the E. coli isolates, consistent effort is required to investigate and monitor the resistant genes.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.9.1-9.11
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• 2022

Background: Escherichia coli, which causes subclinical or clinical mastitis in cattle, is responsible for transmitting antimicrobial resistance via human consumption of raw milk or raw milk products. Objectives: The objective of this study was to investigate the molecular characteristics of 183 E. coli from bulk tank milk of five different dairy factories in Korea. Methods: The molecular characteristics of E. coli such as serogroup, virulence, antimicrobial resistance, and integron genes were detected using polymerase chain reaction and antimicrobial susceptibility were tested using the disk diffusion test. Results: In the distribution of phylogenetic groups, group D was the most prevalent (59.6%) and followed by group B1 (25.1%). The most predominant serogroup was O173 (15.3%), and a total of 46 different serotypes were detected. The virulence gene found most often was fimH (73.2%), and stx1, fimH, incC, fyuA, and iutA genes were significantly higher in isolates of phylogenetic group B1 compared to phylogenetic groups A, B2, and D (p < 0.05). Among 64 E. coli isolates that showed resistance to at least one antimicrobial, the highest resistance rate was observed for tetracyclines (37.5%). All 18 integron-positive E. coli carried the integron class I (int1) gene, and three different gene cassette arrangements, dfrA12+aadA2 (2 isolates), aac(6')-Ib3+aac(6')-Ib-cr+aadA4 (2 isolates), and dfrA17+aadA5 (1 isolate) were detected. Conclusions: These data suggest that the E. coli from bulk tank milk can be an indicator for dissemination of antimicrobial resistance and virulence factors via cross-contamination.

• Biomedical Science Letters
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• v.18 no.1
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• pp.79-82
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• 2012

Acinetobacter infections are of great concern in clinical settings because of multi-drug resistance (MDR) and high mortality of the infected patients. The MDR Acinetobacter baumannii has emerged as a significant infectious agent in hospitals worldwide. The purpose of this study was to determine for molecular characterization of MDR A. baumannii clinical isolates obtained from the Wonju Christian Hospital in Gangwon province of Korea. A total of seventy nonduplicate A. baumannii isolates were collected from the Wonju Christian Hospital in Korea from March to April in 2011. All of the MDR A. baumannii isolates were encoded by $bla_{OXA-23-like}$ gene and all isolates with the $bla_{OXA-23-like}$ gene had the upstream element ISAba1 to promote increased gene expression and subsequent resistance to carbapenem. 16S rRNA methylase gene (armA) was detected in 44 clinical isolates which were resistant to amikacin, and phosphotransferase genes encoding aac(3)-Ia and aac(6')-Ib were the most prevalent. A combination of 16S rRNA methylase and aminoglycoside-modifying enzyme genes (armA, aac(3)-Ia, aac(6')-Ib, and aph(3')-Ia) were found in 31 isolates. The sequencing results for the quinolone resistance-determining region (QRDR) of gyrA and parC revealed the presence of Ser (TCA) 83 Leu (TTA) and Ser (TCG) 80 Leu (TTG) substitutions in the respective enzymes for all MDR. Molecular typing for MDR A. baumannii could be helpful in confirming the identification of a common source or cross-contamination. This is an important step in enabling epidemiological tracing of these strains.

• Korean Journal of Veterinary Service
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• v.40 no.1
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• pp.27-33
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• 2017

Salmonella enterica subsp. enterica is a common microbial flora in pet turtles, which could opportunistically become pathogenic to human. Their possession of aminoglycoside resistance genes has important significance both in humans and animal medicine. In this study, twenty-one Salmonella enterica subsp. enterica were isolated from thirty-five individual turtles purchased from pet shops and online markets in Korea. In order to characterize the aminoglycoside susceptibility patterns, antimicrobial susceptibility tests were performed against gentamicin, amikacin and kanamycin of aminoglycoside antimicrobial group. Each of the isolates showed susceptibility to all tested aminoglycosides in disk diffusion and minimum inhibitory concentration (MIC) tests. PCR assay was carried out to determine aminoglycoside resistance genes, integron and integron mediated aminoglycoside genes. None of the isolates showed aac(3)-IIa, aac-(6')-Ib, armA, aphAI-IAB aminoglycoside resistance genes. Only, five isolates (24%) harbored class 1 integron related IntI1 integrase gene. The results suggest that Salmonella enterica subsp. enterica strains isolated from pet turtles are less resistance to aminoglycosides and don't harbor any aminoglycosides resistance genes.