Characterization of pABVA01, a Plasmid Encoding the OXA-24 Carbapenemase from Italian Isolates of
            <i>Acinetobacter baumannii</i>

D’Andrea, Marco Maria; Giani, Tommaso; D’Arezzo, Silvia; Capone, Alessandro; Petrosillo, Nicola; Visca, Paolo; Luzzaro, Francesco; Rossolini, Gian María

doi:10.1128/aac.00178-09

Cited by 91 publications

(94 citation statements)

References 34 publications

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“…Plasmid DNA from E. coli TOP10 (pAb25) was used as the template for sequencing the 3.59-kb sequence encompassing the bla OXA-253 gene by primer walking, i.e., by using primers sequentially designed on the obtained sequence from the inside of the bla OXA-253 gene to the outside. Surprisingly, the genetic environment of the bla OXA-253 gene, with the open reading frames (ORFs) tonB, ORF2, ORF1, repAci2, and repB, differed greatly from that of the bla OXA-143 gene described previously (2) but showed 95% identity with that of the bla OXA-24/-40 gene of pABVA01 plasmid (GR2 group) from an A. baumannii strain from Italy (8), although the bla OXA-253 gene shares only 88% nucleotide identity with the bla OXA-24/-40 gene (Fig. 2).…”

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confidence: 89%

OXA-253, a Variant of the Carbapenem-Hydrolyzing Class D β - Lactamase OXA-143 in Acinetobacter baumannii

Girlich

Damaceno

Oliveira

et al. 2014

Antimicrob Agents Chemother

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The carbapenem-hydrolyzing class D ␤-lactamase OXA-253 was identified in an Acinetobacter baumannii clinical isolate belonging to sequence type 113 (ST113) in Brazil. OXA-253 shares 93.8% amino acid identity with OXA-143. The bla OXA-253 gene is located on a ca. 20-kb plasmid. The genetic environment of the bla OXA-253 gene shares the highest identity with ubiquitous GR2 group plasmids usually carrying bla OXA-24/-40 genes.A cinetobacter baumannii strains are frequently associated with nosocomial infections, and their resistance to carbapenems has risen in recent decades (1, 2). The main mechanisms of resistance to carbapenems of A. baumannii are efflux pumps, porin mutations, overexpression of the chromosomally encoded OXA-51-like ␤-lactamase, and the biosynthesis of acquired carbapenem-hydrolyzing class D ␤-lactamases (CHDLs) (oxacillinases) (3). To date, five main groups of CHDLs have been identified in A. baumannii, the intrinsic chromosomally encoded OXA-51-like enzyme and the acquired chromosomally encoded and plasmidencoded OXA-23-like, OXA-40-like, OXA-58-like, and OXA-143-like enzymes (3). OXA-143 was first described in 2009 from an A. baumannii strain isolated in Brazil in 2004 (2). In this study, we report an A. baumannii clinical strain producing OXA-253, a variant of OXA-143. The genetic environment of the plasmidborne bla OXA-253 gene is detailed.The clinical isolate A. baumannii 25 was recovered from a perineal swab in the intensive care unit in Minas Gerais Hospital, Brazil. MICs were determined by Etest (bioMérieux, La Balmeles-Grottes, France). Isolate 25 was resistant to all ␤-lactams, including carbapenems, and had reduced susceptibility to cefepime according to the CLSI guidelines (4) ( Table 1). It was also resistant to fluoroquinolones (ciprofloxacin), and susceptible to aminoglycosides (gentamicin, tobramycin, amikacin, and netilmicin), and tetracycline. A modified version of the CarbaNP test (5) and the CarbAcinetoNP test (P. Nordmann, personal communication) was performed and showed a carbapenemase activity. PCR experiments, performed as previously described for screening of bla , bla , bla OXA-58 and bla OXA-143 genes (6), identified a bla OXA-143 -like gene. Further sequencing of the PCR product identified the bla OXA-253 gene (GenBank number KC479324) and showed 95% nucleotide identity with the bla OXA-143 gene. The deduced protein sequence showed 17 amino acid differences between OXA-143 and OXA-253 (93.8% identity) (Fig. 1). Those amino acid changes do not occur in the conserved residues STFK (position 70 to 73) and KSG (position 216 to 218) of class D ␤-lactamases or in the FGN structural element (position 144 to 146) of carbapenem-hydrolyzing class D ␤-lactamases (CHDLs) (Fig. 1).Plasmid extraction from A. baumannii 25 performed by the Kieser technique yielded two plasmids of ca. 70 and 20 kb (7). The gene encoding the OXA-253 determinant from A. baumannii 25 was transferred to Escherichia coli TOP10 by electroporation. The resulting E. coli TOP10 strain harboring the plasmid o...

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confidence: 89%

OXA-253, a Variant of the Carbapenem-Hydrolyzing Class D β - Lactamase OXA-143 in Acinetobacter baumannii

Girlich

Damaceno

Oliveira

et al. 2014

Antimicrob Agents Chemother

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“…Analysis of the nucleotide sequence surrounding a plasmid-located bla OXA-40 gene identified the presence of two conserved inverted repeats on either side of the gene, separated from one another by a 6-bp variable region (102). These sequences are homologous to the target sites for XerC and XerD recombinases, which are usually involved in converting chromosome and plasmid dimers into monomers (103).…”

Section: Xerc/xerdmentioning

confidence: 99%

OXA β-Lactamases

2014

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SUMMARY The OXA β-lactamases were among the earliest β-lactamases detected; however, these molecular class D β-lactamases were originally relatively rare and always plasmid mediated. They had a substrate profile limited to the penicillins, but some became able to confer resistance to cephalosporins. From the 1980s onwards, isolates of Acinetobacter baumannii that were resistant to the carbapenems emerged, manifested by plasmid-encoded β-lactamases (OXA-23, OXA-40, and OXA-58) categorized as OXA enzymes because of their sequence similarity to earlier OXA β-lactamases. It was soon found that every A. baumannii strain possessed a chromosomally encoded OXA β-lactamase (OXA-51-like), some of which could confer resistance to carbapenems when the genetic environment around the gene promoted its expression. Similarly, Acinetobacter species closely related to A. baumannii also possessed their own chromosomally encoded OXA β-lactamases; some could be transferred to A. baumannii , and they formed the basis of transferable carbapenem resistance in this species. In some cases, the carbapenem-resistant OXA β-lactamases (OXA-48) have migrated into the Enterobacteriaceae and are becoming a significant cause of carbapenem resistance. The emergence of OXA enzymes that can confer resistance to carbapenems, particularly in A. baumannii , has transformed these β-lactamases from a minor hindrance into a major problem set to demote the clinical efficacy of the carbapenems.

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“…The ST2 3990 strain contained two plasmids, p1-ABST2 (63,320 bp) carrying a complete tra locus and p2-ABST2 (21,846 bp) carrying one copy of the carbapenem-hydrolyzing oxacillinase (CHDL) bla OXA-58 gene, homologous to plasmids pACICU2 and pACICU1, respectively (11). The ST25 strain contained two distinct plasmids, p1-ABST25 (15,267 bp) and p2-ABST25 (8,970 bp), that both carried one copy of the CHDL bla OXA-72 gene and were homologous to plasmids carrying the bla OXA-24 gene (5,12). The single plasmid p1-ABST78 (26,411 bp) identified in the ST78 strain contained one copy of the bla OXA-58 gene and was homologous to plasmids p2-ABST2 and pACICU1 (11).…”

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Genome Sequences of Three Acinetobacter baumannii Strains Assigned to the Multilocus Sequence Typing Genotypes ST2, ST25, and ST78

et al. 2011

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Acinetobacter baumannii is an emerging opportunistic Gram-negative pathogen responsible for hospitalacquired infections. A. baumannii epidemics described in Europe and worldwide were caused by a limited number of genotypic clusters of multidrug-resistant strains. Here, we report the availability of draft genome sequences for three multidrug-resistant A. baumannii strains assigned to multilocus sequence typing genotypes ST2, ST25, and ST78 that were more frequently isolated during outbreaks occurred in Greece, Italy, Lebanon, and Turkey.Acinetobacter baumannii is an emerging opportunistic pathogen, causing a variety of nosocomial infections (13). Outbreaks of A. baumannii were caused by few genotypic clusters of strains that were initially named European clones I, II, and III and are now regarded as international (7, 10, 13) and referred to as ST1, ST2, and ST3, respectively, according to multilocus sequence typing (7). Epidemiological studies showed the prevalence of the international clone II lineage during the last few years (7,8,10,13), as well as the occurrence of epidemics caused by multidrug-resistant strains belonging to novel genotypes ST25 and ST78 in several Mediterranean hospitals (8, 9).The whole-genome sequences of six ST1 (1, 2, 15) and individual ST2, ST3, and ST77 A. baumannii strains were available to date (2,11,14,15). Here, we announce the availability of three draft genome sequences for carbapenem-resistant A. baumannii ST2 strain 3990, ST78 strain 3909, and ST25 strain 4190, isolated during cross-transmission episodes that occurred at the Monaldi Hospital, Naples, Italy, during 2006Italy, during , 2007Italy, during , and 2009.The genomes were sequenced to at least 10-fold coverage using 454 FLX Titanium emPCR pyrosequencing (Roche) according to the manufacturer's recommendations. Draft genomes were assembled using Newbler and automatically annotated using the xBASE2 bacterial genome annotation service (4). The draft genome sequences of the ST2, ST25, and ST78 strains, respectively, consisted of 96, 396, and 236 contigs, comprised 4,015,011 bases, 4,032,291 bases, and 3,954,832 bases, and generated 3,806, 3,910, and 3,721 protein coding sequences by automated annotation against the A. baumannii AB0057 genome. Comparative analysis of the ST2, ST25, and ST78 genomes with the ACICU, AB0057, ABAYE, and ATCC17978 genomes using Mauve software (6) identified 3,068 homologous protein coding sequences at the same relative positions in the seven genomes. Sixty-three DNA segments, ranging in size from 3 to 126 kb, were present only in some of the seven genomes and were either missing or replaced by nonhomologous DNA sequences in others. Consistent with their close genetic relatedness, both the ST2 3990 and the ACICU strain carried a 15.4-kb region containing antimicrobial resistance genes inserted in the ATPase gene locus. At the corresponding chromosomal location, a 12.7-kb region flanked by transposases but devoid of resistance genes was identified in the ST78 but not in the ST25 strain.Complete pl...

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Characterization of pABVA01, a Plasmid Encoding the OXA-24 Carbapenemase from Italian Isolates of Acinetobacter baumannii

Cited by 91 publications

References 34 publications

OXA-253, a Variant of the Carbapenem-Hydrolyzing Class D β - Lactamase OXA-143 in Acinetobacter baumannii

OXA-253, a Variant of the Carbapenem-Hydrolyzing Class D β - Lactamase OXA-143 in Acinetobacter baumannii

OXA β-Lactamases

Genome Sequences of Three Acinetobacter baumannii Strains Assigned to the Multilocus Sequence Typing Genotypes ST2, ST25, and ST78

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