• Korean Journal of Clinical Laboratory Science
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• v.54 no.3
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• pp.179-191
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• 2022

Carbapenem-resistant Enterobacteriaceae (CRE) poses an increasing public health threat and has limited treatment options with high associated mortality. Genotypes of carbapenemase that threaten public health (bla_KPC, bla_NDM, bla_IMP, and bla_VIM) and bla_OXA-48-like genes were detected by phenotypic and molecular diagnosis, and related gene distribution patterns were investigated. Phenotypic testing using the modified Hodge test confirmed positivity in all 41 strains examined, and carbapenemase inhibitory testing using meropenem+phenyl boronic acid or meropenem+EDTA confirmed positivity in 18 and 8 strains, respectively. Polymerase chain reaction revealed the presence of amplification products in 28 strains of bla_KPC, 25 strains of bla_NDM, 5 strains of bla_IMP, 1 strain of bla_VIM, and 13 bla_OXA-48-like strains. In addition, 7 strains of bla_KPC+bla_NDM, 1 strain of bla_KPC+bla_IMP, 1 strain of bla_NDM+bla_OXA-48-like, 1 strain of bla_NDM+bla_VIM, 4 strains of bla_KPC+bla_NDM+bla_IMP, and 4 strains of bla_KPC+bla_NDM+bla_OXA-48-like were identified. Melting curve analysis using real-time PCR was wholly consistent with PCR results. The study shows genetic identification of highly specific CRE by real-time PCR could be used to provide early diagnoses and infection control, improve surveillance, and prevent the transmission of CRE.

• Korean Journal of Microbiology
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• v.50 no.2
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• pp.101-107
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• 2014

Community-acquired antimicrobial resistant bacteria are an emerging problem world-wide. In Korea, resistant bacteria are more prevalent than in other industrialized countries. The aim of this study was to investigate the isolation frequency of methicillin-resistant staphylococci (MRS), Pseudomonas, and Enterobacteriaceae from surrounding environment (home, colleges, public transportation system and possessions) in Seoul, and to examine the level of drug resistance to 13 antimicrobial agents, which are in wide spread clinical use in Korea, as well as new agent, tigecycline in Enterobacteriaceae isolates. Of total 239 samples, 18 (7.5%) MRS, 10 (4.2%) Pseudomonas, and 30 (12.6%) Entarobacteriaceae were isolated. A total of 5 (2.1%) methicillin-resistant S. aureus (MRSA) were detected in home (2 samples), colleges (1 sample), and et cetera (2 sample). A total of 5 (2.1%) Escherichia coli were detected in in home (1 samples), public transportations (3 sample), and et cetera (1 sample). Resistance to cephalosporins, fluoroquinolones, carbapenems, ${\beta}$-lactams, tetracyclines, and aminoglycosides was found in 71.9%, 71.9%, 68.8%, 68.8%, 50.0%, and 25.0% of 32 Enterobacteriaceae isolates, respectively. Also, resistance rate to trimethoprim/sulfamethoxazole of the isolates was a 43.8%. Moreover, 59.4% of the isolates were resistant to new agent, tigecycline and resistance to all agents tested was observed in 3 isolates. Five E. coli isolates were resistant to most of the agents tested, but some of them were susceptible to ciprofloxacin and gentamicin. This study can serve as a data point for future comparisons of possible changes in antibiotic resistance levels in surrounding environment. And multilateral strategies for preventing the incidence and spread of antibiotic resistance are needed.

• 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.

• YAKHAK HOEJI
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• v.41 no.5
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• pp.615-621
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• 1997

Dehydropeptidase-I (DHP-I) was solubilized from porcine kidney by treatment with n-butanol and partially purified 19.25 fold by $(NH_4)_2SO_4$ precipitation, DEAE-Sepharose CL-6B ion exchange chromatography and Sephacryl S-300 HR chromatography with an overall yield of 19.16. DHP-I showed its optimal activity at pH 7.5 and 25$^{\circ}C$. Its activity was stable under neutral and alkaline conditions, but was disappeared under acidic condition. And DHP-I was heat-labile and its activity remained at 45$^{\circ}C$ for 3hrs. The enzyme was not inhibited by dicationic ions, while its activity was increased by $Co^{2+}$(1mM) and $Zn^{2+}$ (0.1mM). The enzyme was inhibited by EDTA and N-ethylmaleimide. The relative molecular mass of DHP-I was estimated to be approximately 100kDa by gel filtration chromatography. The $K_m$ value of DHP-I for glycyldehydrophenylalanine (GDHP) was 1.98mM. DWP20418 [(1R, 5S, 6S)-6-[1-(R)-Hydroxyethyl]-1-methyl-2-[(2S, 4S)-2-(piperazinylcarbonyl)-1-(R)-hydroxyethyl)pyrrolidine-4-thio]carbapen-2-em-3-carboxylic acid], compared with meropenem (MEPM), was rather easily hydrolized by DHP-I, while it was four times more resistant than imipenem (IPM) to DHP-I.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1711-1715
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• 2017

In this study, we characterized the $bla_{NDM-5}$-bearing plasmid in a Klebsiella pneumoniae isolate that had lost the plasmid during serial passage. We determined the complete sequences of the plasmid pCC1410-2, which was extracted from a K. pneumoniae ST709 isolate collected at a Korean hospital from which two NDM-5-producing K. pneumoniae isolates were subsequently isolated. As a result, the pCC1410-2 plasmid had a backbone structure that was similar to those of two plasmids previously reported from the same hospital, but lacked some antibiotic resistance genes ($bla_{TEM-1}$, rmtB, mphR(A), mrx(A), and mph(A)). A 9-bp repeating unit encoding three amino acids (Gln-Gln-Pro) was inserted in TraD in pCC1410-2. Thus, the pCC1410-2 plasmid might be transferred from the previously identified carbapenem-resistant K. pneumoniae, but some delections and inversions might have occurred during the process. We compared the transfer frequency and stability of the plasmids. The relative frequency of conjugative transfer and stability in the host were significantly lower in pCC1410-2 than in previously reported $bla_{NDM-5}$-bearing plasmids in Korea. A low transfer frequency and instability in the host may cause underestimation of carbapenemase-producing Enterobacteriaceae in the clinical setting and in surveillance studies.

• Pediatric Infection and Vaccine
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• v.18 no.1
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• pp.23-30
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• 2011

Purpose : Acinetobacter baumannii is an aerobic, gram negative coccobacillus. Due to its pathogenicity and ability to accumulate diverse mechanisms of resistance, the importance of this organism is increasing. Many reports have targeted adults, and studies of pediatric patients are limited. This study aims to investigate the current status of A. baumannii infection in children. Methods : From January 2001 to December 2008, 505 patients hospitalized with A. baumannii infection were enrolled. Admission records for underlying disease, duration of hospitalization, previous antibiotic use, location of admission, presence of ventilator care, and resistance to antibiotics were retrospectively reviewed and analyzed. Results : Hemato-oncological disease and neurological disease were 30.6% and 24.3% of all cases; therefore, these were the most common underlying diseases of patients with A. baumannii infection. Prevalence of A. baumannii infection was 78.1% in patients with previous antibiotic use, which was higher than that of the group not using previous antibiotic. And prevalence of multi-drug resistant and pan-drug resistant A. baumannii infection was 76.4% and 38.3% in patients with ICU care, 76.8% and 38.9% with ventilator care, and these were higher than the others. Rate of resistance to all groups of antibiotics showed a gradual increase to over 50% in 2008. Multi-drug resistant A. baumannii was 63.5% and pan-drug resistant A. baumannii was 48.2% of all cases. Conclusion : Prevalence of A. baumannii infection and resistance to antibacterial agents of A. baumannii is increasing. Adequate use of antibiotics and infection control should be emphasized in pediatric patients.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.1
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• pp.49-59
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• 2021

This study investigated the use of real-time PCR melting curves for the diagnosis of blaKPC and blaNDM genes among the most frequently detected carbapenemase-producing Enterobacteriaceae in Korea. As a means of addressing the shortcomings of phenotype tests and conventional PCR. The modified Hodge test confirmed positivity in 25 of 35 strains, and carbapenemase inhibition testing confirmed positivity in 14 strains by meropenem+PBA or meropenem+EDTA. PCR analysis showed amplification products in 25 strains of Klebsiella pneumoniae carbapenemases (KPC), 10 of K. pneumoniae, 5 of E. coli, 5 of A. baumannii, 4 of P. aeruginosa, and 1 of P. putida. New Delhi metallo β-lactamase (NDM) identified amplification products in 8 strains, that is, 2 K. pneumoniae, 3 E. coli, 1 P. aeruginosa, 1 E. cloacae, and 1 P. retgeri strains. Real-time PCR melting curve analysis confirmed amplification in 25 strains of KPC and 8 strains of NDM, and these results were 100% consistent with PCR results. In conclusion, our findings suggest early diagnosis of carbapenem resistant Enterobacteriaceae by real-time PCR offers a potential means of antibacterial management that can prevent and control nosocomial infection spread.