• 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 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.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.442-444
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• 2018

Recently, we isolated a multidrug-resistant Escherichia coli strain KBN10P04869 from a patient with acute myeloid leukemia. We report the complete genome of this strain which consists of 5,104,264 bp with 4,457 protein-coding genes, 88 tRNAs, and 22 rRNAs, and the co-occurrence of multidrug- resistant genes including $^{bla}CMY-2$, $^{bla}TEM-1$, $^{bla}CTX-M-15$, $^{bla}NDM-5$, and $^{bla}OXA-18$.

• 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.

• Journal of Life Science
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• v.25 no.12
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• pp.1399-1407
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• 2015

Klebsiella pneumoniae is found in the normal flora of the skin, mouth, respiratory tract, urinary tract, and intestines in human. However, the stain is opportunistic pathogen, which is the causative agent of community acquired pneumonia. Corn silk has been known to be effective for antimicrobial activity against pathogenic bacteria, including K. pneumoniae, Staphylococcus aureus, Bacillus subtilis, Shigella spp., Salmonella spp., Escherichia coli, Pseudomonas aeruginosa, et al. In this study we focused on the antimicrobial properties of con silk water extract of K. penumoniae. K. pneumoniae isolates K. pneumoniae ATCC 13883 and broad-spectrum β-lactamase (BSBL), exteded-spectrum β-lactamase (ESBL), carbapenemase-producers. Antimicrobial susceptibilities were determined by the disk diffusion method. Searches for bla genes were performed by PCR amplication and direct sequencing. MacConkey agar plate medium was prepared using the corn silk extracts (50% or 100%) instead of distilled water for antimicrobial activity test. The microbial growth inhibitory potential of K. pneumoniae was determined by using the MacConkey agar plate spreading method, and the plate was incubated 18 hr at 37℃. Genes encoding β-lactamases including SHV-1 (n=8), SHV-2a (n=8), SHV-5 (n=2), SHV-11 (n=2), SHV-12 (n=18), TEM-1 (n=10), CTX-M-3 (n=2), CTX-M-14 (n=2), CTX-M-15 (n=1), GES-5 (n=5), KPC-2 (n=6), KPC-3 (n=4), and NDM-1 (n=2) were detected. The corn silk extract showed significantly antimicrobial activity against K. pneumoniae ATCC 13883, but BSBLs, ESBLs, and carbapenemase producers were not. Therefore, corn silk extract is thought to be able to assist in the prevention and rapid recovery of infectious disease caused by K. pneumoniae.