PCR Typing of Genetic Determinants for Metallo-β-Lactamases and Integrases Carried by Gram-Negative Bacteria Isolated in Japan, with Focus on the Class 3 Integron

Shibata, Naohiro; Doi, Yohei; Yamane, Kunikazu; Yagi, Tetsuya; Kurokawa, Hiroshi; Shibayama, Keigo; Kato, Haru; Kai, Kumiko; Arakawa, Yoshichika

doi:10.1128/jcm.41.12.5407-5413.2003

Cited by 297 publications

(210 citation statements)

References 27 publications

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“…PCR screening was performed for OXA-type carbapenemases (bla , 24 , 51 and -58-like ) and MBLs (bla IMP-1 , IMP-2 and VIP-2-like ) (Shibata et al, 2003;Turton et al, 2006).…”

Section: Methodsmentioning

confidence: 99%

Diverse carbapenem-resistance mechanisms in 16S rRNA methylase-producing Acinetobacter baumannii

Yamada

Suwabe

2013

Journal of Medical Microbiology

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Multidrug-resistant Acinetobacter baumannii has become a global problem. This study characterized amikacin-resistant A. baumannii isolated from eight patients during April 2010-March 2011 in our university hospital and examined the possible mechanisms in three cases in which carbapenem susceptibility changed to carbapenem resistance during treatment of the patients. The armA gene, which is one of the 16S rRNA methylase genes and is associated with high MICs of aminoglycosides, was positive in all isolates. The bla OXA-51 gene and ISAba1 and ISAba1-bla ADC were positive in all isolates, but ISAba1-bla OXA-51 was positive in only three isolates. The CarO outer-membrane protein was lost in one isolate. In the first case, both the susceptible and the resistant isolates were positive for ISAba1-bla OXA-51 , but the expression of the bla ADC gene was increased 3.1-fold in the carbapenem-resistant isolate of the pair. In the second case, the carbapenem-resistant strain became positive for ISAba1-bla OXA-51 , resulting in 21.5-fold increased expression of bla OXA-51 , compared to the carbapenem-susceptible strain of the pair. In the third case, the carbapenemase genes remained negative despite the carbapenem resistance, but the expression of the adeB gene was increased 4.6-fold after acquisition of carbapenem resistance. Multilocus sequence typing analysis of two isolates showing representative pulsed-field gel electrophoresis patterns demonstrated that both isolates were classified to sequence type 2 (ST2). These results showed that the 16S rRNA methylaseproducing A. baumannii, initially susceptible to carbapenem, acquired carbapenem resistance via diverse mechanisms.

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“…PCR screening was performed for OXA-type carbapenemases (bla , 24 , 51 and -58-like ) and MBLs (bla IMP-1 , IMP-2 and VIP-2-like ) (Shibata et al, 2003;Turton et al, 2006).…”

Section: Methodsmentioning

confidence: 99%

Diverse carbapenem-resistance mechanisms in 16S rRNA methylase-producing Acinetobacter baumannii

Yamada

Suwabe

2013

Journal of Medical Microbiology

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“…Screening for production of metallo-β-lactamase was performed using the metallo-β-lactamase inhibitor (21). Strains were inoculated onto MullerHinton agar plates where two Kerby-Bauer disks, one containing 30 µg of ceftazidime and one containing 3 mg of sodium mercaptoacetic acid (SMA) (Eiken), 272 M. OHARA ET AL were placed at a distance of 15-20 mm from each other.…”

Section: Methodsmentioning

confidence: 99%

Molecular Characterization of Imipenem‐Resistant Pseudomonas aeruginosa in Hiroshima, Japan

Ohara

Kouda

Onodera

et al. 2007

Microbiology and Immunology

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Nosocomial infections by Pseudomonas aeruginosa continue to be a widespread problem in Asia, North and South America, and Europe. Carbapenems are the most potent β-lactams against P. aeruginosa because its strong affinity to penicillin binding proteins, stability against most serine β-lactamases and high permeability across the outer membrane (7,8). However, intensive use of the carbapenems facilitated the emergence of carbapenem-resistant P. aeruginosa. Low level resistance to imipenem (MIC, 8 to 32 µg/ml) in P. aeruginosa is mostly due to reduced uptake of the drug as a result of loss of the porin OprD (8). This mechanism correlates with continued expression of AmpC β-lactamase. Resistance to meropenem but not imipenem may also arise via overexpression of the MexA-MexBOprM efflux pump (14). High level resistance to carbapenems (MICϾ32 µg/ml) can be caused by the presence of metallo-β-lactamase, which exhibits a broadspectrum resistance to β-lactams including carbapenems. Metallo-β-lactamase-producing P. aeruginosa Abstract: Pseudomonas aeruginosa showing resistance to imipenem were found in 100 of 1,058 strains (9.5%) from six hospitals (a-f) in Hiroshima City, Japan. Of the 100 strains, 14 (14%) were double disk synergy test positive using sodium mercaptoacetic acid disks, and 18 (18%) were bla IMP-1 or blaVIM-2 allele positive by polymerase chain reaction (PCR). Among 100 imipenem-resistant strains, 32 were categorized into multi-drug resistant strains, in which 13 were positive for the metallo-␤-lactamase gene. Fifty-one strains (51%) among the 100 imipenem-resistant strains had elevated RND efflux pump activity against levofloxacin. But only 6 of 51 strains were classified as multi-drug resistant strains. The pulsed field gel electrophoresis analysis of the SpeI-digested DNA from the 100 isolates suggested not only clonal spread but spread of heterogeneous clones started to contribute to the prevalence of metallo-␤-lactamase producing P. aeruginosa strains in Japanese hospitals.

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“…MBL-encoding genes are usually found as gene cassettes in class 1 integrons (Shibata et al, 2003). Typical class 1 integrons contain two conserved segments (CSs), the 59-CS and the 39-CS (Stokes & Hall, 1989;Bennett, 1999).…”

Section: Introductionmentioning

confidence: 99%

Relevance of resistance levels to carbapenems and integron-borne bla IMP-1, bla IMP-7, bla IMP-10 and bla VIM-2 in clinical isolates of Pseudomonas aeruginosa

Zhao

Chen

Ito

et al. 2009

Journal of Medical Microbiology

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Relevance of resistance levels to carbapenems and integron-borne bla IMP-1 , bla IMP-7 , bla IMP-10 and bla Molecular detection and surveillance of the resistance genes harboured by Pseudomonas aeruginosa are becoming increasingly important in assessing and controlling spread and colonization in hospitals, and in guiding the treatment of infections. This study analysed the resistance mechanisms of carbapenem-resistant clinical isolates of P. aeruginosa and identified the associated integron-borne metallo-b-lactamase (MBL)-encoding genes. Twenty-seven imipenem (IPM)-resistant clinical isolates of P. aeruginosa were divided into three groups according to their resistance levels to carbapenems. Strains bearing bla IMP-10 showed extremely high-level resistance to IPM, with MICs of 512-2048 mg ml "1 . By comparison, strains bearing bla IMP-1 , bla IMP-7 and bla VIM-2 showed an intermediate level of resistance, with MICs of 32-256 mg ml "1 . The non-MBL-producing strains showed a low level of resistance, with MICs of 8-32 mg ml "1 . The same trend in resistance levels was also observed when resistance to other carbapenems, such as meropenem and panipenem, was determined. DNA sequencing showed that the MBL-encoding gene cassettes were carried by class 1 integrons. The bla IMP-1 , bla IMP-7 and bla IMP-10 gene cassettes were preceded by a hybrid P ant promoter, TGGACA-N 17 -TAAACT, and the bla VIM-2 gene cassette was preceded by a weak promoter, TGGACA-N 17 -TAAGCT. Most of the MBL-encoding genes were linked to one or two resistance genes encoding aminoglycoside-modifying enzymes, such as aac(69)Iae, aac(69)II, aacA7, aacC4, aadA1, aadA2 and aadA6, highlighting the multidrug-resistant properties of these clinical isolates.

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PCR Typing of Genetic Determinants for Metallo-β-Lactamases and Integrases Carried by Gram-Negative Bacteria Isolated in Japan, with Focus on the Class 3 Integron

Cited by 297 publications

References 27 publications

Diverse carbapenem-resistance mechanisms in 16S rRNA methylase-producing Acinetobacter baumannii

Diverse carbapenem-resistance mechanisms in 16S rRNA methylase-producing Acinetobacter baumannii

Molecular Characterization of Imipenem‐Resistant Pseudomonas aeruginosa in Hiroshima, Japan

Relevance of resistance levels to carbapenems and integron-borne bla IMP-1, bla IMP-7, bla IMP-10 and bla VIM-2 in clinical isolates of Pseudomonas aeruginosa

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