• Title/Summary/Keyword: 16S rRNA methylase

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Molecular Characterization and Prevalence of 16S Ribosomal RNA Methylase Producing Bacteria in Amikacin Resistant Gram-negative Bacilli Isolated from Clinical Specimens

  • Shin, Kyung-A;Hwang, Seock-Yeon;Hong, Seung-Bok
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.299-306
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    • 2012
  • Recently, the prevalence of 16S rRNA methylase conferring high-level resistance to aminoglycosides has been increasing in Gram-negative bacilli globally. We determined the prevalence and genotype of these methylase-producing bacteria, and characterized the co-resistance to ${\beta}$-lactam antibiotics and quinolone in Gram-negative clinical isolates collected in 2010 at a hospital in Korea. Among 65 amikacin-resistant isolates screened from 864 Gram-negative bacilli (GNB), 16S rRNA methylase genes were detected from 49 isolates, including Acinetobacter baumannii (43), Klebsiella pneumoniae (2), Proteus mirabilis (2) and Serratia marcescens (1), Empedobacter brevis (1). All of the 16S rRNA methylase genotype was armA and no variant sequences of amplified PCR products for armA were noted. The 16S rRNA methylase producing bacteria showed much higher resistance to aminoglycoside for Enterobacteriaceae and glucose non-fermenting (NF)-GNB and to imipenem for glucose NF-GNB, than the non-producing isolates. All of the 16S rRNA methylase producing Enterobacteriaceae had the extended-spectrum-${\beta}$-lactamase. In addition, two K. pneumoniae concurrently produced both plasmid-mediated AmpC ${\beta}$-lactamase and qnrB gene. All of the amikacin-resistant A. baumannii (43) co-harbored armA 16S rRNA methylase and $bla_{OXA-23}$ carbapenemase. In conclusion, 16S rRNA methylase producing bacteria were very prevalent among GNB in South Korea, and were commonly associated with co-resistance, including carbapenem and quinolone.

Prevalence and molecular characteristics of 16s rRNA methylase gene rmtB in amikacin resistant Escherichia coli isolated from South Korea

  • Belaynehe, Kuastros Mekonnen;Won, Ho Geun;Yoon, In Joong;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.59 no.3
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    • pp.157-160
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    • 2019
  • The production of rmtB-encoded 16S rRNA methylases has emerged as a novel mechanism promoting high-level resistance toward aminoglycosides in Gram-negative bacteria. Between 2015 and 2017, 636 distinct commensal Escherichia (E.) coli isolates were collected from different farms in South Korea to determine the prevalence and molecular characteristics of rmtB. The positive rates of rmtB between all the isolates and amikacin-resistant isolates were 1.1 and 100%, respectively. High-level aminoglycoside resistance could be transferred by conjugation from rmtB-positive donors to higher amikacin-resistance efficacies. This is the first report of 16S rRNA methylase-encoding genes in E. coli isolated from food-producing animals in Korea.

Cloning and Sequencing of Resistance Determinants to Aminoglycoside Antibiotics from Sterptoalloteichus hindustanus ATCC 31219 (Streptoalloteichus hindustanus ATCC 31219로부터 아미노글라이코사이드계 항생제에 내성을 지정하는 유전자의 클로닝 및 염기서열 결정)

  • Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Eom, Joon-Ho;Yoon, Sung-Joon;Hyun, Chang-Gu;Suh, Joo-Won
    • Microbiology and Biotechnology Letters
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    • v.23 no.4
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    • pp.384-389
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    • 1995
  • Streptoalloteichus hindustanus ATCC 31219, a nebramycin complex producer, is similar to Streptomyeces tenebrarius in a viewpoint of resistance to a wide range of aminoglycoside antibiotics. S. tenebrarius has resistance mechanisms of 16s rRNA methylation and aminogycoside modification. However, it is not known whether resistance mechanisms of Stall. hindustanus are the same as in S. tenebrarius. Therefore, we have tried to isolate resistance determinants from Stall. hindustanus. Two different types of aminoglycoside resistance determinants were isolated from Stall. hindustanus and expressed in Streptomyces lividans TK24. The apramycin resistance gene (amr) and the tobramycin resistance gene (tmr) isolated from Stall. hindustanus showed broad resistance spectrum against a dozen of aminoglycoside antibiotics. The complete nucleotide sequences of apramycin resistance gene (amr) were determined. The deduced amino acid sequence of the amr gene of Stall hindustanus ATCC 31219 showed extensive sequence homology to the 16s rRNA methylase gene (kamB) of S. tenebrarius.

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Multidrug-Resistant Providencia Isolates Carrying $bla_{PER-1},\;bla_{VIM-2}$, and armA

  • Lee, Hee-Woo;Kang, Hee-Young;Shin, Kyeong-Seob;Kim, Jung-Min
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.272-274
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    • 2007
  • During May to July 2004, three strains of Providencia spp. with multidrug-resistance (MDR) were isolated from urinary specimen of three patients hospitalized with a same hospital room. By PCR analysis, all three strains have been found to carry both VIM-2 type $metallo-{\beta}-lactamase$ gene and PER-1 type extended spectrum ${\beta}-lactamase$ gene. One out of three strains carried additional resistance gene, armA, 16S rRNA methylase gene responsible for high level resistance to aminoglycosides. To our knowledge, this is the first report on the identification of Providencia spp. simultaneously carrying $bla_{VIM-2},\;bla_{PER-1}$, and armA genes.

Cloning and Sequence Analysis of the Aminoglycoside Resistance Gene from a Nebramycin Complex Producer, Streptoalloteichus hindustanus

  • Hyun, Chang-Gu;Kim, Jong-Woo;Han, Jae-Jin;Choi, Young-Nae;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.146-151
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    • 1998
  • The aminoglycoside multiple-resistance determinant from Streptoalloteichus hindustanus was cloned into Streptomyces lividans and named nbrB. The 1.2-kb ApaI- BclI fragment encompassing nbrB was located within a 2.6-kb ApaI fragment by successive subcloning experiments. The complete DNA nucleotide sequence of 1.2-kb containing nbrB was determined. The sequence contains an open reading frame that putatively encodes a polypeptide of 281 amino acids with a predicted molecular weight of 30,992. The deduced amino acid sequence of nbrB shows identities of 85.1% to kgmB of S. tenebrarius, 59.6% to sgm of Micromonospora zionensis, and 57.7% to grm of M. rosea. The similarity of nbrB to kgmB suggests that nbrB encodes a 16S rRNA methylase similar to that encoded by kgmB and that both genes might be derived from a common ancestral gene.

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Prevalence of Multi-drug Resistant Acinetobacter baumannii Producing OXA-23-like from a University Hospital in Gangwon Province, Korea

  • Jang, In-Ho;Lee, Gyu-Sang;Choi, Il;Uh, Young;Kim, Sa-Hyun;Park, Min;Woo, Hyun-Jun;Choi, Yeon-Im;Kim, Jong-Bae
    • 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.

Genetic Organization of a 50-kb Gene Cluster Isolated from Streptomyces kanamyceticus for Kanamycin Biosynthesis and Characterization of Kanamycin Acetyltransferase

  • ZHAO XIN QING;KIM KYOUNG ROK;SANG LI WEI;KANG SUK HO;YANG YOUNG YELL;SUH JOO WON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.346-353
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    • 2005
  • A 50-kb chromosome DNA region was isolated from Streptomyces kanamyceticus by screening the fosmid genomic library, using the 16S rRNA methylase gene (kmr) as a probe. Sequence analysis of this region revealed 42 putative open reading frames (ORFs), which included biosynthetic genes such as genes responsible for 2-deoxystreptamine (2­DOS) biosynthesis as well as genes for resistance and regulatory function. Also, the kanamycin acetyltransferase gene (kac) was characterized by in vitro enzyme assay, which conferred E. coli BL21 (DE3) with 10, 50, and 80-times higher resistance to kanamycin A, tobramycin, and amikacin, respectively, than the control strain had, thus strongly indicating that the isolated gene cluster is very likely involved in kanamycin biosynthesis. This work provides a solid basis for further elucidation of the kanamycin biosynthesis pathway as well as the productivity improvement and construction of new hybrid antibiotics.

An Analysis of the Antibiotic Resistance Genes of Multi-Drug Resistant (MDR) Acinetobacter baumannii (다제내성 Acinetobacter baumannii 의 항생제 내성 유전자 분석)

  • Lim, Jina;Lee, Gyusang;Choi, Yeonim;Kim, Jongbae
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.217-224
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    • 2016
  • Acinetobacter baumannii (A. baumannii) is prevalent in hospital environments and is an important opportunistic pathogen of nosocomial infection. It is known that this pathogen cause herd infection in hospitals, and the mortality rate is remarkably higher for patients infected with this pathogen and already have other underlying diseases. Herein, we investigated the antibiotic resistance rate and the type of resistance genes in 85 isolates of multi-drug resistant A. baumannii from the samples commissioned to laboratory medicine in two university hospitals-in hospital A and hospital B-located in Cheonan and Chungcheong provinces, respectively, in Korea. As a result, $bla_{OXA-23-like}$ and $bla_{OXA-51-like}$ were detected in 82 stains (96.5%). These 82 strains of $bla_{OXA-23-like}$ producing A. baumannii were confirmed with the ISAba1 gene found at the top of the $bla_{OXA-23-like}$ genes by PCR, inducing the resistance against carbapenemase. The armA, AME gene that induces the resistance against aminoglycoside was detected in 34 strains out of 38 strains from Hospital A (89.5%), and in 40 strains out of 47 strains from Hospital B (85.1%), while AMEs were found in 33 strains out of 38 strains from Hospital A (70.2%) and in 44 strains out of 47 strains in Hospital B (93.6%). Therefore, it was found that most multi-drug resistant A. baumannii from the Cheonan area expressed both acethyltransferase and adenyltransferase. This study investigated the multi-drug resistant A. baumannii isolated from Cheonan and Chungcheong provinces in Korea, and it is thought that the results of the study can be utilized as the basic information to cure multi-drug resistant A. baumannii infections and to prevent the spread of drug resistance.