Presence of Genes for .BETA.-Lactamases of Two Different Classes on a Single Plasmid from a Clinical Isolate of Serrtia marcescens.

Yano, Hisakazu; Kuga, Akio; Irinoda, Kazuhiko; Okamoto, Ryoichi; Kobayashi, Toshimitsu; Inoue, Matsuhisa

doi:10.7164/antibiotics.52.1135

Cited by 9 publications

(9 citation statements)

References 16 publications

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“…The bacterial strains and plasmids used in this study are listed in Table 1. In 1996, Serratia marcescens KU3838 was isolated from the urine of a patient with a urinary tract infection at a hospital in northern Japan (41) and was found to contain plasmid pKU501. Escherichia coli K12 ML4901 (18) was used as the recipient for plasmids.…”

Section: Methodsmentioning

confidence: 99%

“…Isolation of plasmid DNA using the small-scale alkaline method (33) and gene amplification by PCR (29,41) were performed as previously described. Primer pairs P1 (5Ј-CGG ATG AAG GCA CGA ACC CA-3Ј) and P2 (5Ј-AAG CAG ACT TGA CCT GAT AGT-3Ј) (TaKaRa Shuzo Co., Ltd. Kyoto, Japan), which were chosen on the basis of the published pMS350 integron and 3Ј-conserved region sequence (17), were used in the PCR experiments.…”

Section: Methodsmentioning

confidence: 99%

“…Assay of ␤-lactamase. Enzyme activity was determined by spectrophotometry (UV2000; Shimazu Corp., Tokyo, Japan) at 30°C in 50 mM phosphate buffer (pH 7.0) with 10 M ZnCl 2 (41). One unit of enzyme activity was defined as the amount of enzyme hydrolyzing 1 mol of substrate in 1 min at 30°C.…”

Section: Methodsmentioning

confidence: 99%

“…Previously, we described pKU501 as a novel plasmid carrying not only bla IMP but also the genes for TEM-1-type ␤-lactamases as well as producing both types of ␤-lactamases (41). A strain carrying pKU501 was more resistant to meropenem and panipenem, especially meropenem, than to imipenem.…”

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confidence: 99%

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Plasmid-Encoded Metallo-β-Lactamase (IMP-6) Conferring Resistance to Carbapenems, Especially Meropenem

Yano

Kuga

Okamoto

et al. 2001

Antimicrob Agents Chemother

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134

122

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In 1996, Serratia marcescens KU3838 was isolated from the urine of a patient with a urinary tract infection at a hospital in northern Japan and was found to contain the plasmid pKU501. Previously, we determined that pKU501 carries bla IMP and the genes for TEM-1-type ␤-lactamases as well as producing both types of ␤-lactamases (H. Yano, A. Kuga, K. Irinoda, R. Okamoto, T. Kobayashi, and M. Inoue, J. Antibiot. 52:1135-1139, 1999). pKU502 is a recombinant plasmid that contains a 1.5-kb DNA fragment, including the metallo-␤-lactamase gene, and is obtained by PCR amplification of pKU501. The sequence of the metallo-␤-lactamase gene in pKU502 was determined and revealed that this metallo-␤-lactamase gene differed from the gene encoding IMP-1 by one point mutation, leading to one amino acid substitution: 640-A in the base sequence of the IMP-1 gene was replaced by G, and Ser-196 was replaced by Gly in the mature enzyme. This enzyme was designated IMP-6. The strains that produced IMP-6 were resistant to carbapenems. The MICs of panipenem and especially meropenem were higher than the MIC of imipenem for these strains. The k cat /K m value of IMP-6 was about sevenfold higher against meropenem than against imipenem, although the MIC of meropenem for KU1917, which produced IMP-1, was lower than that of imipenem, and the MIC of panipenem was equal to that of imipenem. These results support the hypothesis that IMP-6 has extended substrate profiles against carbapenems. However, the activity of IMP-6 was very low against penicillin G and piperacillin. These results suggest that IMP-6 acquired high activity against carbapenems, especially meropenem, via the point mutation but in the process lost activity against penicillins. Although IMP-6 has reduced activity against penicillins due to this point mutation, pKU501 confers resistance to a variety of antimicrobial agents because it also produces TEM-1-type enzyme.Carbapenems such as imipenem, panipenem, and meropenem are the most potent agents for treatment of gramnegative infections due to the stability of these agents against the majority of ␤-lactamases and their high rate of permeation through bacterial outer membranes. In addition, imipenem, panipenem, and meropenem have a high affinity for penicillinbinding protein (PBP) 2 of gram-negative bacteria except for Pseudomonas aeruginosa. In P. aeruginosa, imipenem and panipenem have a high affinity for PBP 2 and meropenem has an affinity for PBP 3 (6, 19, 40).However, some clinical isolates have been reported to be resistant to carbapenems due to production of metallo-␤-lactamases. There have been increasing reports, especially from Japan (7,14,21,27,35), of gram-negative bacteria that carry the transferable carbapenem resistance gene bla IMP , including isolates of P. aeruginosa and Serratia marcescens. The metallo-␤-lactamases are class B enzymes in Ambler's molecular classification (1) and belong to Bush group 3 (3,31). Most of these metallo-␤-lactamases confer resistance not only to carbapenems but also to other ␤-lactams an...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Plasmid-Encoded Metallo-β-Lactamase (IMP-6) Conferring Resistance to Carbapenems, Especially Meropenem

Yano

Kuga

Okamoto

et al. 2001

Antimicrob Agents Chemother

Self Cite

134

122

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“…Real-time PCR assays were developed in this study for detection of CTX-M-1-type and SHV-type ␤-lactamases, while TEM and OXA PCR were performed according to the methods of Yano et al (20) and Ouellette et al (12), respectively. SHV primers (SHV1, 5Ј-CTTTATCGGCCCTCACTC AA-3Ј; SHV2, 5Ј-AGGTGCTCATCATGGGAAAG-3Ј) were designed on the basis of the published SHV-1 sequence (15), and CTX-M-1-type primers (CM1, 5Ј-CGTCACGCTGTTGT TAGGAA-3Ј; CM2, 5Ј-TCGGTTCGCTTTCACTTTTC-3Ј) were based on the published sequences of CTX-M-1 (1), CTX-M-3 (4), CTX-M-10 (11), CTX-M-11 (GenBank accession no.…”

mentioning

confidence: 99%

Molecular Epidemiological Analysis of Escherichia coli Isolates Producing Extended-Spectrum β-Lactamases for Identification of Nosocomial Outbreaks in Stockholm, Sweden

Fang

Lundberg²,

Olsson‐Liljequist

et al. 2004

J Clin Microbiol

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A Klebsiella pneumoniae producing three kinds of class A β-lactamases encoded by one single plasmid isolated from a patient in Huashan Hospital, Shanghai, China

Xiong

Zhu

Wang

et al. 2004

International Journal of Antimicrobial Agents

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Presence of Genes for .BETA.-Lactamases of Two Different Classes on a Single Plasmid from a Clinical Isolate of Serrtia marcescens.

Cited by 9 publications

References 16 publications

Plasmid-Encoded Metallo-β-Lactamase (IMP-6) Conferring Resistance to Carbapenems, Especially Meropenem

Plasmid-Encoded Metallo-β-Lactamase (IMP-6) Conferring Resistance to Carbapenems, Especially Meropenem

Molecular Epidemiological Analysis of Escherichia coli Isolates Producing Extended-Spectrum β-Lactamases for Identification of Nosocomial Outbreaks in Stockholm, Sweden

A Klebsiella pneumoniae producing three kinds of class A β-lactamases encoded by one single plasmid isolated from a patient in Huashan Hospital, Shanghai, China

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