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      Molecular epidemiology and antimicrobial resistance patterns of carbapenem-resistant Acinetobacter baumannii isolates from patients admitted at ICUs of a teaching hospital in Zunyi, China

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          Abstract

          Background

          Carbapenem-resistant Acinetobacter baumannii (CRAB) has emerged as a predominant strain of healthcare-associated infections worldwide, particularly in intensive care units (ICUs). Therefore, it is imperative to study the molecular epidemiology of CRAB in the ICUs using multiple molecular typing methods to lay the foundation for the development of infection prevention and control strategies. This study aimed to determine the antimicrobial susceptibility profile, the molecular epidemiology and conduct homology analysis on CRAB strains isolated from ICUs.

          Methods

          The sensitivity to various antimicrobials was determined using the minimum inhibitory concentration (MIC) method, Kirby-Bauer disk diffusion (KBDD), and E-test assays. Resistance genes were identified by polymerase chain reaction (PCR). Molecular typing was performed using multilocus sequence typing (MLST) and multiple-locus variable-number tandem repeat analysis (MLVA).

          Results

          Among the 79 isolates collected, they exhibited high resistance to various antimicrobials but showed low resistance to levofloxacin, trimethoprim-sulfamethoxazole, and tetracyclines. Notably, all isolates of A. baumannii were identified as multidrug-resistant A. baumannii (MDR-AB). The bla OXA-51-like, adeJ, and adeG genes were all detected, while the detection rates of bla OXA-23-like (97.5%), adeB (93.67%), bla ADC (93.67%), qacEΔ1-sul1 (84.81%) were higher; most of the Ambler class A and class B genes were not detected. MLST analysis on the 79 isolates identified five sequence types (STs), which belonged to group 3 clonal complexes 369. ST1145 Ox was the most frequently observed ST with a count of 56 out of 79 isolates (70.89%). MLST analysis for non-sensitive tigecycline isolates, which were revealed ST1145 Ox and ST1417 Ox as well. By using the MLVA assay, the 79 isolates could be grouped into a total of 64 distinct MTs with eleven clusters identified in them. Minimum spanning tree analysis defined seven different MLVA complexes (MCs) labeled MC1 to MC6 along with twenty singletons. The locus MLVA-AB_2396 demonstrated the highest Simpson’s diversity index value at 0.829 among all loci tested in this study while also having one of the highest variety of tandem repeat species.

          Conclusion

          The molecular diversity and clonal affinities within the genomes of the CRAB strains were clearly evident, with the identification of ST1144 Ox, ST1658 Ox, and ST1646 Oxqaq representing novel findings.

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          Most cited references56

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          Global optimal eBURST analysis of multilocus typing data using a graphic matroid approach

          Background Multilocus Sequence Typing (MLST) is a frequently used typing method for the analysis of the clonal relationships among strains of several clinically relevant microbial species. MLST is based on the sequence of housekeeping genes that result in each strain having a distinct numerical allelic profile, which is abbreviated to a unique identifier: the sequence type (ST). The relatedness between two strains can then be inferred by the differences between allelic profiles. For a more comprehensive analysis of the possible patterns of evolutionary descent, a set of rules were proposed and implemented in the eBURST algorithm. These rules allow the division of a data set into several clusters of related strains, dubbed clonal complexes, by implementing a simple model of clonal expansion and diversification. Within each clonal complex, the rules identify which links between STs correspond to the most probable pattern of descent. However, the eBURST algorithm is not globally optimized, which can result in links, within the clonal complexes, that violate the rules proposed. Results Here, we present a globally optimized implementation of the eBURST algorithm – goeBURST. The search for a global optimal solution led to the formalization of the problem as a graphic matroid, for which greedy algorithms that provide an optimal solution exist. Several public data sets of MLST data were tested and differences between the two implementations were found and are discussed for five bacterial species: Enterococcus faecium, Streptococcus pneumoniae, Burkholderia pseudomallei, Campylobacter jejuni and Neisseria spp.. A novel feature implemented in goeBURST is the representation of the level of tiebreak rule reached before deciding if a link should be drawn, which can used to visually evaluate the reliability of the represented hypothetical pattern of descent. Conclusion goeBURST is a globally optimized implementation of the eBURST algorithm, that identifies alternative patterns of descent for several bacterial species. Furthermore, the algorithm can be applied to any multilocus typing data based on the number of differences between numeric profiles. A software implementation is available at .
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            Multilocus sequence typing of bacteria.

            Multilocus sequence typing (MLST) was proposed in 1998 as a portable, universal, and definitive method for characterizing bacteria, using the human pathogen Neisseria meningitidis as an example. In addition to providing a standardized approach to data collection, by examining the nucleotide sequences of multiple loci encoding housekeeping genes, or fragments of them, MLST data are made freely available over the Internet to ensure that a uniform nomenclature is readily available to all those interested in categorizing bacteria. At the time of writing, over thirty MLST schemes have been published and made available on the Internet, mostly for pathogenic bacteria, although there are schemes for pathogenic fungi and some nonpathogenic bacteria. MLST data have been employed in epidemiological investigations of various scales and in studies of the population biology, pathogenicity, and evolution of bacteria. The increasing speed and reduced cost of nucleotide sequence determination, together with improved web-based databases and analysis tools, present the prospect of increasingly wide application of MLST.
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              PHYLOViZ: phylogenetic inference and data visualization for sequence based typing methods

              Background With the decrease of DNA sequencing costs, sequence-based typing methods are rapidly becoming the gold standard for epidemiological surveillance. These methods provide reproducible and comparable results needed for a global scale bacterial population analysis, while retaining their usefulness for local epidemiological surveys. Online databases that collect the generated allelic profiles and associated epidemiological data are available but this wealth of data remains underused and are frequently poorly annotated since no user-friendly tool exists to analyze and explore it. Results PHYLOViZ is platform independent Java software that allows the integrated analysis of sequence-based typing methods, including SNP data generated from whole genome sequence approaches, and associated epidemiological data. goeBURST and its Minimum Spanning Tree expansion are used for visualizing the possible evolutionary relationships between isolates. The results can be displayed as an annotated graph overlaying the query results of any other epidemiological data available. Conclusions PHYLOViZ is a user-friendly software that allows the combined analysis of multiple data sources for microbial epidemiological and population studies. It is freely available at http://www.phyloviz.net.
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                Author and article information

                Contributors
                Role: Role: Role: Role:
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                URI : https://loop.frontiersin.org/people/1617707Role: Role:
                URI : https://loop.frontiersin.org/people/222835Role: Role: Role:
                Journal
                Front Cell Infect Microbiol
                Front Cell Infect Microbiol
                Front. Cell. Infect. Microbiol.
                Frontiers in Cellular and Infection Microbiology
                Frontiers Media S.A.
                2235-2988
                01 December 2023
                2023
                : 13
                : 1280372
                Affiliations
                [1] 1 Department of Laboratory Medicine, The First People’s Hospital of Zunyi (The Third Affiliated Hospital of Zunyi Medical University) , Zunyi, China
                [2] 2 Scientific Research Center, The First People’s Hospital of Zunyi (The Third Affiliated Hospital of Zunyi Medical University) , Zunyi, China
                Author notes

                Edited by: Alessandro Russo, Magna Græcia University, Italy

                Reviewed by: Ariadnna Cruz-Córdova, Federico Gómez Children’s Hospital, Mexico; Reem Mostafa Hassan, Cairo University, Egypt

                *Correspondence: Kaifeng Wu, kiphoonwu@ 123456126.com ; He Zha, zhahe666@ 123456126.com

                †These authors have contributed equally to this work

                Article
                10.3389/fcimb.2023.1280372
                10722174
                38106474
                e0ee22a9-febf-4e0c-9a5c-037f77c09a7f
                Copyright © 2023 Xiong, Deng, Yang, Shen, Chen, Tian, Zha and Wu

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 20 August 2023
                : 13 November 2023
                Page count
                Figures: 3, Tables: 3, Equations: 0, References: 56, Pages: 11, Words: 4961
                Funding
                The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The project received support from the Guizhou High-level (BAI) Innovative Talents Project (QIANKehe Platform & Talents-GCC[2022]042-1), the National Natural Science Foundation of China (No.81760475), the Innovation Group Project provided by Guizhou Provincial Department of Education (QianJiaoheKYzi [2021]019), the Key Discipline Project of Clinical Laboratory Diagnostics funded by Guizhou Provincial Health Commission (QianWeijianhan[2021]160), the Key Discipline Project of Clinical Laboratory Diagnostics funded by Zunyi Municipal Health Bureau (2022-1444), the Zunyi United Science and Technology Fund Project (Zunyi Kehe HZzi [2018]159), the Science and Technology Fund Project of Guizhou Provincial Health Commission (gzwkj2021-367), and Guizhou Provincial Basic Research Program(Natural Science) (No.ZK[2023]488).
                Categories
                Cellular and Infection Microbiology
                Original Research
                Custom metadata
                Clinical Microbiology

                Infectious disease & Microbiology
                carbapenem-resistant acinetobacter baumannii (crab),molecular epidemiology,antibiotic resistance,intensive care units (icus),multilocus sequence typing(mlst),multiple-locus variable-number tandem repeat analysis(mlva)

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