PCR detection of metallo-beta-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum beta-lactams

Senda, Kazuyoshi; Arakawa, Yoshichika; Ichiyama, Satoshi; Nakashima, Kiyotaka; Ito, Hideo; Ohsuka, Shinji; Shimokata, Kaoru; Kato, Naoya; Ohta, Michio

doi:10.1128/jcm.34.12.2909-2913.1996

Cited by 370 publications

(149 citation statements)

References 17 publications

(21 reference statements)

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“…The presence of metallo-b-lactamases (MBLs) was determined using MBL Etests (AB Biodisk). PCRs for MBLs were performed using primers and PCR conditions described previously [26,27]. The VIM consensus primers were those recommended by Poirel et al [27].…”

Section: Detection Of Carbapenem Resistance Genesmentioning

confidence: 99%

Emergence and spread of carbapenem-resistant strains of Acinetobacter baumannii in a tertiary-care hospital in Poland

Wróblewska

Towner

Marchel

et al. 2007

Clinical Microbiology and Infection

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This study analysed the occurrence of carbapenem resistance among Acinetobacter baumannii isolates from a tertiary-care hospital in Poland, together with the molecular epidemiology of these isolates and the risk-factors for their acquisition and possible nosocomial spread. The medical charts of 21 patients with Acinetobacter infection or colonisation revealed that A. baumannii isolates were obtained most frequently from intensive care unit and surgical patients (particularly those receiving transplantation surgery). First isolation occurred, on average, on day 21 following admission (range 5-45 days). Infection with Acinetobacter contributed directly to the death of seven patients. Several patients were infected with more than one strain, and molecular typing revealed the co-circulation of three predominant clones, of which two belonged to the Acinetobacter lineages designated as European clones I and II. All three clones encoded an OXA-51-type carbapenemase, but were negative for carbapenemases belonging to the OXA-23, OXA-24 and OXA-58 families. The OXA-51 gene was found in both resistant and susceptible isolates, and was not associated directly with carbapenem resistance. Etests with imipenem and imipenem plus EDTA indicated production of a metallo-beta-lactamase (MBL) in carbapenem-resistant isolates. PCRs for IMP-type MBLs were negative, but PCR using consensus primers for VIM-type MBLs were positive for carbapenem-resistant isolates belonging to the European clone II lineage. The occurrence of a VIM-type MBL in association with one of the epidemic lineages of A. baumannii is a cause for concern. Further studies are needed to evaluate possible inter-hospital spread of resistant A. baumannii strains in Poland.

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Section: Detection Of Carbapenem Resistance Genesmentioning

confidence: 99%

Emergence and spread of carbapenem-resistant strains of Acinetobacter baumannii in a tertiary-care hospital in Poland

Wróblewska

Towner

Marchel

et al. 2007

Clinical Microbiology and Infection

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“…The PCR method described by Senda et al [2] which ampli¢es a 587-bp fragment (internal bla IMP gene) was used as screen for the presence of bla IMP . In addition the primers 5P-ATC CAA GCA GCA AGC GCG TTA-3P (IMP13) and 5P-CGT GCT GCT GCA ACG ACT TGT-3P (IMP15), which are a modi¢cation of those described by Chu et al [7] to ensure the ampli¢cation of an 879-bp fragment which contained the bla IMP open reading frame (ORF), were used for sequencing.…”

Section: Detection Of Bla Imp Type Genes and Nucleotide Sequence Analmentioning

confidence: 99%

“…The appearance therefore 10 years ago of transferable R plasmids carrying the IMP-1 L-lactamase conferring resistance to all carbapenems and almost all other L-lactam antibiotics was regarded as a serious event [1]. The bla IMPÀ1 gene has subsequently dispersed amongst at least 12 species of Gram-negative bacteria in Japan, a process facilitated by the presence of the bla IMPÀ1 gene within the integron In 31 [2,3]. Integrons enable the movement of gene cassettes such as bla IMPÀ1 amongst di¡erent plasmids (thus circumventing incompatibility and replica-tion barriers) and the microbial genome.…”

Section: Introductionmentioning

confidence: 99%

Occurrence of a new metallo-β-lactamase IMP-4 carried on a conjugative plasmid in Citrobacter youngae from the People's Republic of China

Hawkey¹

2001

FEMS Microbiology Letters

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During the course of an antimicrobial resistance surveillance programme in Guangzhou, the People's Republic of China, single strains of Citrobacter youngae and Pseudomonas aeruginosa were identified which were resistant to imipenem and found to carry the carbapenemase gene bla IMP . PCR screening of the citrobacter strain with specific primers for the bla IMP type genes gave a 587-bp product which when sequenced gave 100% homology with the bla IMPÀ4 sequence reported recently from Acinetobacter spp. The determinant in the C. youngae strain was found to be located on a 156-kb plasmid capable of transfer to Escherichia coli UB1637 by conjugation. Sequencing of the bla IMPÀ4 open reading frame in the C. youngae strain and adjacent sequences not only confirmed the presence of bla IMPÀ4 but also identified that a conserved core site found within the 59-bp element of integrons was present and the same as the one described in the only other occurrence of bla IMPÀ4 in Acinetobacter spp. isolated from an intensive care unit in Hong Kong. This is the second report of transferable carbapenemase genes in Enterobacteriacae outside of Japan and the first in the People's Republic of China. Under the selective pressure of carbapenems and extended spectrum cephalosporins use we might expect this gene to spread and widespread surveillance should be instituted. ß FEMS Microbiology Letters 194 (2001) 53^57 www.fems-microbiology.org Fig. 1. Comparison of the deduced nucleotide sequences of the di¡erent bla IMP genes, identical residues to bla IMPÀ1 are indicated by a dash. P.M. Hawkey et al. / FEMS Microbiology Letters 194 (2001) 53^57

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“…Therefore, these primers ful¢l the important criteria for PCR primer optimization [17]. The primers for bla OXA , bla CMYÀ1 , bla FOX , bla CMY , bla AmpCC , bla LAT , bla BIL , and bla IMI are ¢rstly reported in this paper and are di¡erent from previously reported primers [4,7,8,13].…”

Section: Resultsmentioning

confidence: 79%

“…Nucleotide sequence accession numbers taken from GenBank (Genetic Sequence Database, Bethesda, MD, USA), EMBL (European Molecular Biology Laboratory, Heidelberg, Germany), and/or DDBJ (DNA Data Bank of Japan, Mishima, Japan) databases are shown in Table 1. Other L-lactamases used in the multiple nucleotide sequence alignment are AmpC [12] for bla AmpC (chromosomal AmpC L-lactamase gene from Citrobacter freundii OS60), IMP-1 [13], and IMP-2 [14] for bla IMP . Except for O1, O2, C3, and IN2 of Table 2, the primers were designed by selecting identical sequences in multiple nucleotide alignment of ESBL genes and using the Primer Calculator program (Williamstone Enterprises, Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

Restriction fragment length dimorphism–PCR method for the detection of extended-spectrum β-lactamases unrelated to TEM- and SHV-types

Lee¹

2001

FEMS Microbiology Letters

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The diagnostic ability of the restriction fragment length dimorphism^polymerase chain reaction (RFLD^PCR) method was evaluated. Seven primer pairs, newly designed from 44 L-lactamase genes encoding extended-spectrum L-lactamases not related to TEM-and SHVtypes, were used to differentiate OXA-2, FOX-3, CMY-3, IMP-1, and IMI-1 L-lactamases. The RFLD^PCR was carried out successfully, and these genes were differentiated by the sizes of their PCR products and by the difference in restriction fragment length when each amplicon was digested with a unique restriction enzyme. This discriminatory detection of the genes was confirmed by sequencing the PCR products. ß

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PCR detection of metallo-beta-lactamase gene (blaIMP) in gram-negative rods resistant to broad-spectrum beta-lactams

Cited by 370 publications

References 17 publications

Emergence and spread of carbapenem-resistant strains of Acinetobacter baumannii in a tertiary-care hospital in Poland

Emergence and spread of carbapenem-resistant strains of Acinetobacter baumannii in a tertiary-care hospital in Poland

Occurrence of a new metallo-β-lactamase IMP-4 carried on a conjugative plasmid in Citrobacter youngae from the People's Republic of China

Restriction fragment length dimorphism–PCR method for the detection of extended-spectrum β-lactamases unrelated to TEM- and SHV-types

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