Nosocomial spread of Klebsiella pneumoniae isolates producing blaGES-4 carbapenemase at a Japanese hospital

Yamasaki, Katsutoshi; Komatsu, Masaru; Ono, Tamotsu; Nishio, Hisaaki; Sueyoshi, Noriyuki; Kida, Kaneyuki; Satoh, Kaori; Toda, Hirofumi; Nishi, Isao; Akagi, Masahiro; Mizutani, Tetsu; Nakai, Isako; Kofuku, Tomomi; Orita, Tamaki; Zikimoto, Takuya; Nakamura, Tatsuya; Wada, Yasunao

doi:10.1016/j.jiac.2016.09.006

Cited by 8 publications

(6 citation statements)

References 18 publications

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“…GES enzymes were initially described in Klebsiella pneumoniae in France using a rectal swab from a one-month-old girl who was hospitalized in French Guiana 7 . Since then, bla GES has been reported in several species and in several countries, including Brazil, most frequently in non-fermenting bacilli such as Pseudomonas aeruginosa , Acinetobacter baumannii , and enterobacteria such as K. pneumoniae 8 - 10 . GES is part of a group of ESBL and carbapenemases, comprising 43 variants ( http://bldb.eu/BLDB.php?prot=A#GES ) that have been found in several species of gram-negative bacteria 3 .…”

Section: Discussionmentioning

confidence: 99%

First report of bla GES-1 in Proteus mirabilis clinical isolates

Beltrão

Oliveira

Lópes

2021

Rev. Soc. Bras. Med. Trop.

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Proteus mirabilis is one of the main pathogens causing urinary tract infections and sepsis. To our knowledge, this is the first report of a P. mirabilis hosting bla GES . The presence of these genes was determined using PCR and sequencing. We identified the presence of bla GES-1 in all three isolates. In addition, we identified the bla KPC-2 and bla NDM-1 genes in the two strains. These data emphasize the importance of monitoring and surveillance of all enterobacteria. The circulation of P. mirabilis strains carrying bla GES-1 constitutes a new scenario of resistance in this species and should be an epidemiological alert for global health.

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Section: Discussionmentioning

confidence: 99%

First report of bla GES-1 in Proteus mirabilis clinical isolates

Beltrão

Oliveira

Lópes

2021

Rev. Soc. Bras. Med. Trop.

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“…All 126 strains of CPE isolated from 2001 to 2018 at 18 Japanese medical facilities from Western Japan (17 acute-care hospitals and 1 commercial laboratory) and 168 ESBL-Ec strains extracted randomly were included in the study ( Table 1 ). The 126 strains of CPE were provided in the framework of the Study of Bacterial Resistance in the Kinki Region of Japan (SBRK), of which 37 strains were those used in our previous four studies ( Ohno et al, 2017 , 2020 ; Yamasaki et al, 2017 ; Kuchibiro et al, 2018 ) and 89 strains were used in the present study. Two of the four studies were epidemiological studies of CPE in a primary care hospital in Tenri, Japan, conducted from 2010 to 2015, and these included fecal isolates ( Ohno et al, 2017 , 2020 ).…”

Section: Methodsmentioning

confidence: 99%

“…Two of the four studies were epidemiological studies of CPE in a primary care hospital in Tenri, Japan, conducted from 2010 to 2015, and these included fecal isolates ( Ohno et al, 2017 , 2020 ). Besides, one study conducted an outbreak survey of CPE ( bla GES group) in a primary care hospital in Wakayama City, Japan, in 2009, and the source was blood ( Yamasaki et al, 2017 ). One study was collected for the purpose of epidemiological study of CPE from 2000 to 2016 in 15 facilities in Kinki region, Japan, and these did not include fecal isolates ( Kuchibiro et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

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Genomic Characterization of ESBL- and Carbapenemase-Positive Enterobacteriaceae Co-harboring mcr-9 in Japan

Nakamura

Niki

et al. 2021

Front. Microbiol.

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Worldwide spread of Enterobacteriaceae resistant to colistin, a polypeptide antibacterial drug for last-resort treatment of carbapenemase-producing Enterobacteriaceae (CPE) infections, is concerning. This study aimed to elucidate colistin MICs and molecular characteristics of mcr-1 to mcr-9 of ESBL-producing Escherichia coli (ESBL-Ec) and CPE in Japan and clarify the genomic structure of strains harboring mcr genes (especially mcr-9). This study included 168 ESBL-Ec and 126 CPE strains isolated at Japanese medical facilities. Colistin susceptibility testing and multiplex PCR targeting mcr-1 to mcr-9 were performed for all strains with S1-nuclease pulsed-field gel electrophoresis, Southern blot hybridization, and whole-genome sequencing (WGS) with hybrid assembly performed for mcr gene-carrying strains. Two CPE strains showed a MIC ≥ 4 μg/ml in colistin susceptibility testing, with no known resistance mechanism detected. However, PCR conducted on all target strains detected three mcr-9-carrying strains showing colistin susceptibility. The blaCTX–M–62-positive E. coli THUN648 strain simultaneously carried blaCTX–M–62 and mcr-9 on a 275-kbp plasmid. Besides, blaIMP–6 + blaCTX–M–2-positive Klebsiella pneumoniae THUN262 and blaGES–24-positive Enterobacter kobei THUN627 had mcr-9 encoded on the chromosome. Only THUN627 encoded qseB/C, which is suggested to be a regulatory gene for mcr-9, downstream of mcr-9. However, this strain showed no increased expression of these genes in mRNA quantitative analysis under colistin exposure. Colistin MICs of ESBL-Ec and CPE in Japan were all below 2 μg/ml, which is below the epidemiological cutoff (ECOFF) value (https://eucast.org/) or clinical breakpoint (CB) (CLSI M100-S30) reported for colistin, indicating neither “microbiological” nor “clinical” resistance. Several colistin-susceptible Enterobacteriaceae carrying silent mcr-9 encoded on plasmids and chromosomes have already spread worldwide along with other antimicrobial resistance genes. However, the mechanism of colistin resistance by mcr-9 remains unclear.

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“…The resulting mutant enzyme exhibits carbapenemase activity (Bonnin et al, 2017). In Japan, GES-4 and GES-5 carbapenemase-producing P. aeruginosa have caused severe hospital-acquired infections (Kanayama et al, 2016; Yamasaki et al, 2017). Reports of GES-type enzymes remain rare but are steadily increasing (Naas et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Development of a Novel Loop-Mediated Isothermal Amplification Method to Detect Guiana Extended-Spectrum (GES) β-Lactamase Genes in Pseudomonas aeruginosa

et al. 2019

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Infections caused by multidrug-resistant Pseudomonas aeruginosa in hospitalized patients are often fatal, and nosocomial infections caused by Guiana extended-spectrum (GES) β-lactamase-producing strains are of growing concern. Several genotypes of the GES β-lactamase gene (blaGES) include a single missense mutation, a change from G to A at nucleotide position 493 (G493A) that changes glycine to serine; the mutant enzyme exhibits carbapenemase activity. Rapid and reliable identification of drug-resistance is important in clinical settings; however, culture methods remain the gold standard. Conventional and real-time PCR cannot identify carbapenemase-producing genotypes, and direct DNA sequencing is essential. We established a novel loop-mediated isothermal amplification (LAMP) method to detect various genotypes of blaGES and another LAMP method to discriminate carbapenemase genotypes of blaGES. We evaluated the two assays using clinical P. aeruginosa strains. Two primer sets targeting blaGES (GES-LAMP) and the point mutation (Carba-GES-LAMP) were designed and evaluated for specificity and sensitivity. The detection limit of the GES-LAMP method was assessed using purified DNA and DNA-spiked clinical samples (urine, sputum, and blood). To determine the clinical usefulness of the methods, we used different (genotypically and phenotypically) P. aeruginosa clinical isolates, collected from diverse geographical locations between 2003 and 2012. The novel LAMP assay targeting blaGES was highly specific. The detection limit was 10 DNA copies per reaction; the assay was 10-fold more sensitive than conventional PCR. The LAMP assay detected blaGES with high sensitivity in all DNA-spiked samples; PCR did not detect blaGES in blood samples. The GES-LAMP method correctly detected the 5 isolates containing blaGES among the 14 isolates tested. Using these isolates, we confirmed that our Carba-GES-LAMP method of detecting point mutations correctly identified the two blaGES positive organisms with carbapenemase activity. To the best of our knowledge, this is the first report of the GES β-lactamase gene detection assay using the LAMP method. Our new assays effectively detect blaGES and critical unique mutations.

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Nosocomial spread of Klebsiella pneumoniae isolates producing blaGES-4 carbapenemase at a Japanese hospital

Cited by 8 publications

References 18 publications

First report of bla GES-1 in Proteus mirabilis clinical isolates

First report of bla GES-1 in Proteus mirabilis clinical isolates

Genomic Characterization of ESBL- and Carbapenemase-Positive Enterobacteriaceae Co-harboring mcr-9 in Japan

Development of a Novel Loop-Mediated Isothermal Amplification Method to Detect Guiana Extended-Spectrum (GES) β-Lactamase Genes in Pseudomonas aeruginosa

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