• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.4
• /
• pp.444-452
• /
• 2019

Carbapenem resistance, mediated by the major acquired metallo-β-lactamase (MBL) genes, has been increasingly reported, particularly for clinical isolates of Acinetobacter spp. Of the 191 nonduplicate clinical isolates of the carbapenem-nonsusceptible Acinetobacter spp. evaluated, 125 isolates (65.4%) were positive for the modified imipenem or meropenem-Hodge test, and 49 isolates (25.7%) were positive for the imipenem-EDTA+SMA double disk synergy test (DDS). PCR and sequencing of the bla_VIM-2-allele and bla_IMP-1-allele showed that 29 A. baumannii isolates and 1 A. calcoaceticus isolate had bla_VIM-2, whereas 16 A. baumannii isolates and 2 A. calcoaceticus isolates had bla_IMP-6; 1 isolate of the A. genomospecies 3 had bla_VIM-2 and bla_AIM-1. All the above MBL genes belong to class 1 integron. The size of class 1 integron encompassing bla_VIM-2 or bla_IMP-6 ranges from 2.8 kb to 3.2 kb in clinical isolates of A. baumannii, and 3.2 kb to 3.5 kb in clinical isolates of A. genomospecies 3. bla_VIM-2 was most often located first or second in the class 1 integron, and these integrons often included aacA4. Due to dispersion of the MBL-producing Acinetobacter spp. as well as integron, which may encompass various resistance genes, there is an expectation for the increase of multidrug resistant Gram-negative bacteria, including resistance of carbapenems such as imipenem or meropenem. Hence, the development of new antimicrobial agents for treating severe Acinetobacter spp. infections is needed.

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

• Biomedical Science Letters
• /
• v.25 no.4
• /
• pp.321-330
• /
• 2019

Carbapenem is recently considered as the last resort of the therapeutics for gram negative bacterial infection. Increasing of organisms producing metallo-β-lactamase (MBL), we have difficulty in choosing the antimicrobial agents. Among 345 clinical isolates of Pseudomonas spp., 61 isolates (17.7%) were positive for the modified imipenem or meropenem-Hodge test and 55 isolates (15.9%) were positive for the imipenem-EDTA + SMA double disk synergy test (DDS). PCR and sequencing of bla_VIM-2-allele and bla_IMP-1-allele showed that 17 isolates of Pseudomonas aeruginosa, 9 isolates of Pseudomonas taiwnensis and 2 Pseudomonas plecoglossicida had bla_VIM-2, and 22 isolates of P. aeruginosa and one Pseudomonas otitidis had bla_IMP-6. These MBL genes were all in class 1 integron. The size of class 1 integron with bla_VIM-2 ranged from 3.5 kb to 5.5 kb in clinical isolates of Pseudomonas spp. including P. aeruginosa. bla_VIM-2 was most often located first in the class 1 integron, sometimes in the second or third position, and these integrons often had aacA4 or aadA1. Strict infection control measures are needed to more effectively prevent further spread of these MBL-producing Pseudomonas spp. In addition, MBL-producing Pseudomonas spp. is expected to continue to spread in various countries and regions.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.22 no.6
• /
• pp.413-419
• /
• 2021

The purpose of this study was to investigate the ability to treat and prevent infection by multiple Gram-negative bacterial pathogens as a last choice option in the treatment of serious infections in clinical settings. The global spread of extended-spectrum 𝛽-lactamases (ESBLs) and/or carbapenemases in microorganisms are of enormous concern to health services because they are often associated with multi-drug resistance which significantly restricts the antibiotic treatment options. In this study, the antimicrobial resistance profiles of bacteria isolated from South Korean market-derived meat samples were determined by the disc diffusion method. PCR was used to detect the presence of antibiotic resistance genes and ESBL producing genes. In total, we tested 181 isolated colonies from 36 market-derived meat samples. Single PCR and DNA sequencing results revealed that genes blaVIM, blaBIC, blaKPC, and blaSIM were present in the bacteria isolated from retail meat. The bacteria in the meat were separately sequenced and based on alignment, four different bacteria were identified. These findings suggest that bacteria found in retail meats are a reservoir for the spreading of ESBL blaVIM, blaBIC, blaKPC, and blaSIM resistance genes and bacteria strains.

• Journal of Life Science
• /
• v.22 no.9
• /
• pp.1268-1276
• /
• 2012

The emergence and dissemination of carbapenem-resistant bacteria have resulted in limitations of antibiotic treatment and potential outbreaks of metallo-${\beta}$-lactamase (MBL) producing Pseudomonas aeruginosa resistant to carbapenems. In this study, we conducted molecular characterization of the MBL genes of the ${\beta}$-lactam drug-resistant P. aeruginosa and prepared basic data for treatment and prevention of proliferation of antimicrobial-resistant bacterial infections. Forty-two P. aeruginosa isolates of 254 were resistant to imipenem or meropenem. Among the 42 isolates, 28 isolates were positive for the Hodge test, and 23 isolates were positive for the EDTA-disk synergy test (EDST). MBLs were detected in 59.5% (25/42) of P. aeruginosa isolates. Eight isolates harbored $bla_{IMP-6}$, whereas 17 isolates harbored $bla_{VIM-2}$. The $bla_{IMP-6}$ gene was in a class 1 integron containing five gene cassettes: $bla_{IMP-6}$, qac, aacA4, $bla_{OXA-1}$, and aadA1. Some strains that produce IMP-6 and VIM-2 showed epidemiological relationships. The $bla_{IMP-6}$ gene in carbapenem-resistant P. aeruginosa showed an identical pattern to a gene cassette that was reported at a hospital in Daegu, Korea. Therefore, MBL-producing P. aeruginosa is already endemic in the community. We are concerned that the existence of carbapenem-resistant bacteria containing the blaMBL gene may increase pressure on antibiotic selection when treating infections. We believe that we should select appropriate antibiotics based on the antibiotic susceptibility test and continue the research to prohibit the emergence and spread of antibiotics resistant bacteria.

• Journal of Microbiology
• /
• v.45 no.3
• /
• pp.272-274
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.10
• /
• pp.1733-1737
• /
• 2007

A total of 2,280 nonduplicate clinical isolates of Pseudomonas aeruginosa, obtained nationwide from Korean non-tertiary care hospitals from 2002 to 2005, were identified and their susceptibilities to aminoglycosides, antipseudomonal penicillins, carbapenems, cephalosporins, monobactams, and quinolones were studied, together with their production of ${\beta}$-lactamases. Using disk diffusion and minimum inhibitory concentration tests, it was found that 2.9% of isolates were multidrug-resistant (MDR) P. aeruginosa. An EDTA-disk synergy test, PCR amplification with specifically designed primers, and direct sequencing of the PCR products showed that the $bla_{OXA-10}$, $bla_{VIM-2}$, $bla_{OXA-2}$, $bla_{OXA-17}$, $bla_{PER-1}$, $bla_{SHV-12}$, and $bla_{IMP-1}$ genes were carried by 34.3%, 26.9%, 3.0%,3.0%, 1.5%, 1.5%, and 1.5% of 67 MDR P. aeruginosa isolates, respectively. The prevalence of MDR P. aeruginosa was three-fold higher, compared with that from the United States. More than two types of ${\beta}$-lactamase genes were carried by 10.4% of isolates. The most prevalent ${\beta}$-lactamase genes were $bla_{VIM-2}$ and $bla_{OXA-10}$. This study is the first description of MDR P. aeruginosa trom non-tertiary care hospitals in Korea and the coexistence of the $bla_{VIM-2}$, $bla_{IMP-1}$, or $bla_{PER-1} in these clinical isolates.

• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.4
• /
• pp.428-435
• /
• 2019

The rapid increase and dissemination of carbapene mases, such as Klebsiella pneumoniae carbapenemase (KPC), has become a major problem within the field of healthcare-related infection. There are few antibiotics to treat carbapenem-resistant Enterobacteriaceae (CRE) infections, so the identification of resistant bacterial mechanisms is critical to initiate infection control and conduct epidemiological research. A rapid and effective method for detecting KPC-producing bacteria is needed to avoid therapeutic failures and introduce measures to prevent and control the dissemination of these multi-resistant bacteria. During the study period, 31 isolates (seven isolates of Acinetobacter spp., six isolates of Morganella morganii, five isolates of Pseudomonas aeruginosa, five isolates of Proteus mirabilis, one isolate of Proteus vulgaris, two isolates of Enterobacter cloacae, one isolate of Enterobacter aerogenes, one isolate of Klebsiella pneumoniae, one isolate of Klebsiella oxytoca, one isolate of Serratia marcescens and one isolate of Escherichia coli) were identified by the VITEK. Gram negative rod bacteria were the most frequently isolated from urine (35.5%), blood (19.4%), sputum (16.1%), pus (9.7%), ascitic fluid (9.7%), tracheal aspirates (6.5%) and bile juice (3.2%). Analysis using the PCR method identified the bla_KPC gene in the K. oxytoca1 strain, but the bla_IMP, bla_VIM and bla_OXA-48 genes are not amplified. In conclusion, diagnosis using the PCR method can accurately and quickly diagnose KPC, thus establishing quick preventive measures to prevent the spread of KPC in hospitals.

• Korean Journal of Clinical Laboratory Science
• /
• v.43 no.2
• /
• pp.68-74
• /
• 2011

Pseudomonas aeruginosa is an aerobic, Gram-negative, glucose-nonfermenting bacterium, which has emerged as a serious opportunistic pathogen. Recently, outbreaks of carbapenem resistant P. aeruginosa give rise to significant therapeutic challenges for treating nosocomial infections. The genes of metallo-${\beta}$-lactamase (MBL), a powerful carbapenemase, are carried as a part of the mobile gene cassettes inserted into integrons playing an important role in rapid dissemination of antibiotic resistance genes among bacterial isolates. In this study, we investigated the prevalence of integron in imipenem resistant P. aeruginosa isolates. A total of 61 consecutive, non-duplicate, and imipenem resistant P. aeruginosa strains were isolated from a university hospital in the Chungcheong province of Korea. We employed repetitive extragenic palindromic sequence-based PCR (rep-PCR) method for the selection of clonally different P. aerusinosa strains. PCR and DNA sequencing were conducted for the detection of integrons. Twenty-one clonally different P. aeruginosa strains were isolated. Only one (P28) of the strains harbored $bla_{VIM-2}$ that was found as gene cassettes in class 1 integrons. Four of 21 carbapenem resistant P. aeruginosa strains harbored class 1 integron containing aminoglycoside resistance determinant. All of the integrons detected in the study contained more than one resistance gene cassette, which can mediate resistance to multiple antibiotics. To prevent further spreading of the multi-drug resistant P. aeruginosa, conseguent monitoring and clinical polices are required.

• Korean Journal of Clinical Laboratory Science
• /
• v.42 no.2
• /
• pp.71-80
• /
• 2010

Pseudomonas aeruginosa are important nosocomial pathogens. Their resistance to carbapenem is increasing and causing concerns in Korea. An increasing prevalence of carbapenem resistance mediated by acquired carbapenemase is being reported. Over a 10 month-period from July 2007 to April 2008, 32 strains of imipenem-nonsusceptible P. auruginosa were isolated from Kangwon National University Hospital. To determine the prevalence and genotypes of the carbapenemase-producing clinical isolates, the antibiotic susceptibility was determined by Microscan Walkaway 96 SI System and the carbapenem activity was detected by the modified Hodge test and the imipenem-EDTA-SMA double-disk synergy test. The metallo-${\beta}$-lactamase gene and OXA-type ${\beta}$-lactamase gene reported in Korea were detected by PCR. As for the result of PCR, 30 isolates of P. aeruginosa were found to have $bla_{IMP-1}$-like and 1 isolate was found to have $bla_{IMP-1}$-like and $bla_{IMP-2}$. No clinical isolates were found to have $bla_{SIM-1}$, $bla_{OXA-23}$-like and $bla_{OXA-24}$-like. Random amplified polymorphic DNA (RAPD)-PCR and dendrogram for genetical similarity to band patterns of each clinical isolates were examined. P. aeruginosa were grouped into 7 clusters of up to 50% of similarity index. In the P. aeruginosa group, PS3 was resistant to the most antibiotics, PS1 was susceptible to the most antibiotics. PS7 was resistant to aztreonam unlike other groups. This is the first report of prevalence of carbapenemase in Chuncheon.