Molecular epidemiology of Acinetobacter baumannii spread in an adult intensive care unit under an endemic setting (original) (raw)
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Journal of Clinical Microbiology, 2012
Using a repetitive-sequence-based (rep)-PCR (DiversiLab), we have molecularly typed Acinetobacter nosocomial bloodstream isolates (Acinetobacter baumannii [n ؍ 187], Acinetobacter pittii [n ؍ 23], and Acinetobacter nosocomialis [n ؍ 61]) obtained from patients hospitalized in U.S. hospitals over a 10-year period (1995-2004) during a nationwide surveillance study (Surveillance and Control of Pathogens of Epidemiological Importance [SCOPE]). Patterns of A. baumannii rep-PCR were compared to those of previously identified international clonal lineages (ICs) and were further investigated by multilocus sequence typing (MLST) to compare the two typing methods. Forty-seven of the A. baumannii isolates clustered with the previously defined IC 2. ICs 1, 3, 6, and 7 were also detected. The remaining 81 isolates were unrelated to the described ICs. In contrast, A. pittii and A. nosocomialis isolates were more heterogeneous, as determined by rep-PCR. Our MLST results were in good correlation with the rep-PCR clusters. Our study confirms previous data indicating the predominance of a few major clonal A. baumannii lineages in the United States, particularly IC 2. The presence in the United States of A. baumannii ICs 1, 2, and 3 from as early as 1995 suggests that global dissemination of these lineages was an early event.
Genotyping methods for monitoring the epidemic evolution of A. baumannii strains
Journal of infection in developing countries, 2015
Acinetobacter baumannii is clustered with other phenotypically similar species into what has commonly become known as the ACB complex: A. calcoaceticus, A. pittii and, A. nosocomialis. The ecology and pathology of most of these species are not well understood, mainly because current specific phenotypic techniques have, to date, been insufficient. This has inhibited both the precise identification of, as well as the ability to discriminate between, these clinically important and closely related Acinetobacter strains. However, new genotypic methods have greatly enhanced our capacity to identify the ACB complex. This has resulted in the implementation of more rational infection control programs. Several genotypic identification methods are explored in this study, including non-polymerase chain reaction (PCR)-based and PCR-based methods. These methods include ribotyping, pulsed-field gel electrophoresis, 16S rRNA identification, multilocus sequence typing, single locus sequence typing, ...
Journal of Clinical Microbiology, 2010
This study used a diverse collection of epidemiologically unrelated Acinetobacter baumannii isolates to compare the robustness of a multilocus sequence typing (MLST) scheme, based on conserved regions of seven housekeeping genes, gltA, gdhB, recA, cpn60, rpoD, gyrB, and gpi, with that of sequence-based typing of bla(OXA-51-like) genes (SBT-bla(OXA-51-like) genes). The data obtained by analysis of MLST and SBT-bla(OXA-51-like) genes were compared to the data generated by pulsed-field gel electrophoresis (PFGE). The topologies of the phylogenetic trees generated for the gyrB and gpi genes showed evidence of recombination and were inconsistent with those of the trees generated for the other five genes. MLST identified 24 sequence types (STs), of which 19 were novel, and 5 novel alleles. Clonality was demonstrated by eBURST analysis and standardized index of association values of >1 (P < 0.001). MLST data revealed that all isolates harboring the major bla(OXA-51-like) alleles OXA-66, OXA-69, and OXA-71 fell within the three major European clonal lineages. However, the MLST data were not always in concordance with the PFGE data, and some isolates containing the same bla(OXA-51-like) allele demonstrated <50% relatedness by PFGE. It was concluded that the gyrB and gpi genes are not good candidates for use in MLST analysis and that a SBT-bla(OXA-51-like) gene scheme produced results comparable to those produced by MLST for the identification of the major epidemic lineages, with the advantage of having a significantly reduced sequencing cost and time. It is proposed that studies of A. baumannii epidemiology could involve initial screening of bla(OXA-51-like) alleles to identify isolates belonging to major epidemic lineages, followed by MLST analysis to categorize isolates from common lineages, with PFGE being reserved for fine-scale typing.
Scientific Reports, 2015
Acinetobacter baumannii is a globally distributed nosocomial pathogen that has gained interest due to its resistance to most currently used antimicrobials. Whole genome sequencing (WGS) and phylogenetics has begun to reveal the global genetic diversity of this pathogen. The evolution of A. baumannii has largely been defined by recombination, punctuated by the emergence and proliferation of defined clonal lineages. In this study we sequenced seven genomes from the sequence type (ST)25 lineage and compared them to 12 ST25 genomes deposited in public databases. A recombination analysis identified multiple genomic regions that are homoplasious in the ST25 phylogeny, indicating active or historical recombination. Genes associated with antimicrobial resistance were differentially distributed between ST25 genomes, which matched our laboratorybased antimicrobial susceptibility typing. Differences were also observed in biofilm formation between ST25 isolates, which were demonstrated to produce significantly more extensive biofilm than an isolate from the ST1 clonal lineage. These results demonstrate that within A. baumannii, even a fairly recently derived monophyletic lineage can still exhibit significant genotypic and phenotypic diversity. These results have implications for associating outbreaks with sequence typing as well as understanding mechanisms behind the global propagation of successful A. baumannii lineages. Acinetobacter baumannii is an emergent nosocomial pathogen of increasing interest due to its widespread resistance to antimicrobials 1. A. baumannii is truly a global pathogen, with isolates collected from hospitals around the world 2,3 , including injured soldiers from Iraq 4 and Afghanistan 5. The concern is the emergence of multidrug-resistant (MDR) 6 and extremely drug-resistant (XDR) 7 isolates that are resistant to most currently used therapeutics. Genes that confer resistance in A. baumannii have been documented, including class D beta-lactamases 8 , such as bla OXA-51-like , which appears to be highly conserved across A. baumannii 9. The insertion element ISAba1 is required for carbapenem resistance in bla OXA-51-like positive isolates 10. The genome of A. baumannii is highly plastic 11 , with much of the evolution characterized by recombination 12 and horizontal gene transfer 13. The core genome phylogeny of A. baumannii demonstrates highly divergent genomes, with the emergence of a few highly successful clonal lineages 12,14. While the evolution
Bioscience Journal, 2023
Acinetobacter baumannii is widely recognized in clinical environments due to its infectious capacity, antimicrobial adaptability, and lethality. Analyzing the prevalence of this agent in intra-and extra-hospital environments may reveal target indicators for appropriate management interventions. In this observational cross-sectional study, we evaluated the prevalence of A. baumannii within hospitals with intensive care units and in their external surroundings in a macro-health region of Brazil. Samples of Columba livia (pigeon) droppings from the external environment of four hospitals (n = 40), from floor surfaces (n = 20), and door handles (n = 20) of different hospital wards were collected based on random sampling, all of which were evaluated for the presence of A. baumannii using polymerase chain reactions (PCR). The sensitivity and specificity of the technique was verified after the collected samples were contaminated with clinical samples positive for A. baumannii. We detected a significantly higher A. baumannii prevalence (87.50%, CI = 71.29-100.00) in samples collected within the hospital environment compared with those obtained from the external environment (12.50%, CI = 0.00-28. 71) (p = 0.003). In addition, samples collected from floor surfaces contained bacterial densities (181.3 ± 11.58) that exceeded those in environmental (93.32 ± 1.56) and door handle (142.70 ± 17.14) samples by 94% and 78.71%, respectively. The findings of this study will enhance our understanding of the spatial distribution of A. baumannii and additionally, validate the efficiency of PCR for diagnosis of this infectious agent.
Journal of clinical microbiology, 1994
In 1990, there was a significant increase in the number of lower respiratory tract infections and surgical wound infections in the adult intensive care units of our tertiary care teaching hospital caused by Acinetobacter baumannii compared with the number in 1989. During the 5-month period from April through August 1990, 84 isolates of A. baumannii were recovered from 50 hospitalized patients. Biotyping, comparison of antibiograms, plasmid analysis, and DNA polymorphisms of 20 isolates from 20 different patients, determined by the use of repetitive element PCR with primers aimed at repetitive extragenic palindromic sequences and enterobacterial repetitive intergenic consensus sequences, were used to investigate this apparent outbreak. Biotyping, antibiograms, plasmid analysis, and enterobacterial repetitive intergenic consensus PCR were not useful epidemiologically. Repetitive element PCR-mediated DNA fingerprinting using repetitive extragenic palindromic primers discriminated betwe...
Genome-wide recombination drives diversification of epidemic strains of Acinetobacter baumannii
Proceedings of the National Academy of Sciences of the United States of America, 2011
Acinetobacter baumannii is an emerging human pathogen and a significant cause of nosocomial infections among hospital patients worldwide. The enormous increase in multidrug resistance among hospital isolates and the recent emergence of pandrug-resistant strains underscores the urgency to understand how A. baumannii evolves in hospital environments. To this end, we undertook a genomic study of a polyclonal outbreak of multidrugresistant A. baumannii at the research-based National Institutes of Health Clinical Center. Comparing the complete genome sequences of the three dominant outbreak strain types enabled us to conclude that, despite all belonging to the same epidemic lineage, the three strains diverged before their arrival at the National Institutes of Health. The simultaneous presence of three divergent strains from this lineage supports its increasing prevalence in international hospitals and suggests an ongoing adaptation to the hospital environment. Further genomic comparisons uncovered that much of the diversification that occurred since the divergence of the three outbreak strains was mediated by homologous recombination across 20% of their genomes. Inspection of recombinant regions revealed that several regions were associated with either the loss or swapping out of genes encoding proteins that are exposed to the cell surface or that synthesize cell-surface molecules. Extending our analysis to a larger set of international clinical isolates revealed a previously unappreciated ability of A. baumannii to vary surface molecules through horizontal gene transfer, with subsequent intraspecies dissemination by homologous recombination. These findings have immediate implications in surveillance, prevention, and treatment of A. baumannii infections.
Journal of Medical Microbiology, 2012
We have investigated the reproducibility of DiversiLab rep-PCR fingerprints between two laboratories with the aim of determining if the fingerprints and clustering are laboratory-specific or portable. One-hundred non-duplicate Acinetobacter baumannii isolates were used in this study. DNA isolation and rep-PCR were each performed separately in two laboratories and rep-PCR patterns generated in laboratory A were compared with those from laboratory B. Twelve A. baumannii isolates processed in laboratory A showed ≥98 % pattern similarity with the corresponding 12 isolates tested in laboratory B and were considered identical. Sixty-four isolates showed 95-97.9 % similarity with their corresponding isolates. Twenty-three isolates showed 90-94 % similarity with the corresponding isolates, while one isolate showed only 87.4 % similarity. However, intra-laboratory clustering was conserved: isolates that clustered in laboratory A also clustered in laboratory B. While clustering was conserved and reproducible at two different laboratories, demonstrating the robustness of rep-PCR, interlaboratory comparison of individual isolate fingerprints showed more variability. This comparison allows conclusions regarding clonality to be reached independent of the laboratory where the analysis is performed.
Journal of Clinical Microbiology, 2014
Single-locus bla OXA-51-like sequence-based typing (SBT) was evaluated for its ability to determine correctly sequence types (STs) in Acinetobacter baumannii clinical isolates, in comparison with the Pasteur's multilocus sequence typing (MLST) reference method and 3-locus sequence typing (3-LST). The comparative study was performed in 585 multidrug-resistant (MDR) A. baumannii clinical isolates recovered from 21 hospitals located throughout Greece, Italy, Lebanon, and Turkey. The isolates belonged to nine clonal complexes (CCs) that correspond to 12 distinct sequence types (STs) and to one singleton ST. These clonal lineages predominate worldwide among nosocomial MDR A. baumannii strains. The most common clone was CC2 (ST2 and ST45; n ؍ 278 isolates) followed by CC1 (ST1 and ST20; n ؍ 155), CC25 (n ؍ 65), ST78 (n ؍ 62), CC15 (ST15 and ST84; n ؍ 9), CC10 (n ؍ 4), CC3 (n ؍ 4), CC6 (n ؍ 3), CC54 (n ؍ 3), and CC83 (n ؍ 2). Using the bla OXA-51-like SBT method, all 585 isolates of the study were typed and assigned correctly to the nine CCs and the singleton ST78. The 3-LST method was not able to classify isolates belonging to CC6, CC10, CC54, and CC83, which are not yet characterized in its database. The low-cost and convenient bla OXA-51-like SBT method, compared with 3-LST and MLST, discriminated all epidemic and sporadic lineages of our collection and could be effectively applied to type rapidly A. baumannii strains.
Journal of Clinical Microbiology, 2006
Members of the genus Acinetobacter are ubiquitous in soil and water and are an important cause of nosocomial infections. A rapid method is needed to genotype Acinetobacter isolates to determine epidemiology and clonality during infectious outbreaks. Multilocus PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) is a method that uses the amplicon base compositions to genotype bacterial species. In order to identify regions of the Acinetobacter genome useful for this method, we sequenced regions of six housekeeping genes (trpE, adk, efp, mutY, fumC, and ppa) from 267 isolates of Acinetobacter. Isolates were collected from infected and colonized soldiers and civilians involved in an outbreak in the military health care system associated with the conflict in Iraq, from previously characterized outbreaks in European hospitals, and from culture collections. Most of the isolates from the Iraqi conflict were Acinetobacter baumannii (189 of 216 isolates). Among these, 111 isolates had genotypes identical or very similar to those associated with well-characterized A. baumannii isolates from European hospitals. Twenty-seven isolates from the conflict were found to have genotypes representing different Acinetobacter species, including 8 representatives of Acinetobacter genomospecies 13TU and 13 representatives of Acinetobacter genomospecies 3. Analysis by the PCR/ESI-MS method using nine primer pairs targeting the most information-rich regions of the trpE, adk, mutY, fumC, and ppa genes distinguished 47 of the 48 A. baumannii genotypes identified by sequencing and identified at the species level at least 18 Acinetobacter species. Results obtained with our genotyping method were essentially in agreement with those obtained by pulse-field gel electrophoresis analysis. The PCR/ ESI-MS genotyping method required 4 h of analysis time to first answer with additional samples subsequently analyzed every 10 min. This rapid analysis allows tracking of transmission for the implementation of appropriate infection control measures on a time scale previously not achievable.