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      Antimicrobial susceptibility profile of Neisseria gonorrhoeae from patients attending a medical laboratory, Institut Pasteur de Madagascar between 2014 and 2020: phenotypical and genomic characterisation in a subset of Neisseria gonorrhoeae isolates

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          Abstract

          Objectives

          Antimicrobial-resistant Neisseria gonorrhoeae (NG) is a concern. Little is known about antimicrobial susceptibility profiles and associated genetic resistance mechanisms of NG in Madagascar. We report susceptibility data of NG isolates obtained by the medical laboratory (CBC) of the Institut Pasteur de Madagascar, Antananarivo, Madagascar, during 2014–2020. We present antimicrobial resistance mechanisms data and phenotype profiles of a subset of isolates.

          Methods

          We retrieved retrospective data (N=395) from patients with NG isolated during 2014−2020 by the CBC. We retested 46 viable isolates including 6 found ceftriaxone and 2 azithromycin resistant, as well as 33 isolated from 2020. We determined minimal inhibitory concentrations for ceftriaxone, ciprofloxacin, azithromycin, penicillin, tetracycline and spectinomycin using Etest. We obtained whole-genome sequences and identified the gene determinants associated with antimicrobial resistance and the sequence types (STs).

          Results

          Over the study period, ceftriaxone-resistant isolates exceeded the threshold of 5% in 2017 (7.4% (4 of 54)) and 2020 (7.1% (3 of 42)). All retested isolates were found susceptible to ceftriaxone, azithromycin and spectinomycin, and resistant to ciprofloxacin. The majority were resistant to penicillin (83% (38 of 46)) and tetracycline (87% (40 of 46)). We detected chromosomal mutations associated with antibiotic resistance in gyrA, parC, penA, ponA, porB and mtrR genes. None of the retested isolates carried the mosaic penA gene. The high rate of resistance to penicillin and tetracycline is explained by the presence of bla TEM (94.7% (36 of 38)) and tetM (97.5% (39 of 40)). We found a high number of circulating multilocus STs. Almost half of them were new types, and one new type was among the four most predominant.

          Conclusions

          Our report provides a detailed dataset obtained through phenotypical and genotypical methods which will serve as a baseline for future surveillance of NG. We could not confirm the occurrence of ceftriaxone-resistant isolates. Our results highlight the importance of implementing quality-assured gonococcal antimicrobial resistance surveillance in Madagascar.

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          Most cited references36

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          Trimmomatic: a flexible trimmer for Illumina sequence data

          Motivation: Although many next-generation sequencing (NGS) read preprocessing tools already existed, we could not find any tool or combination of tools that met our requirements in terms of flexibility, correct handling of paired-end data and high performance. We have developed Trimmomatic as a more flexible and efficient preprocessing tool, which could correctly handle paired-end data. Results: The value of NGS read preprocessing is demonstrated for both reference-based and reference-free tasks. Trimmomatic is shown to produce output that is at least competitive with, and in many cases superior to, that produced by other tools, in all scenarios tested. Availability and implementation: Trimmomatic is licensed under GPL V3. It is cross-platform (Java 1.5+ required) and available at http://www.usadellab.org/cms/index.php?page=trimmomatic Contact: usadel@bio1.rwth-aachen.de Supplementary information: Supplementary data are available at Bioinformatics online.
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            Prokka: rapid prokaryotic genome annotation.

            T Seemann (2014)
            The multiplex capability and high yield of current day DNA-sequencing instruments has made bacterial whole genome sequencing a routine affair. The subsequent de novo assembly of reads into contigs has been well addressed. The final step of annotating all relevant genomic features on those contigs can be achieved slowly using existing web- and email-based systems, but these are not applicable for sensitive data or integrating into computational pipelines. Here we introduce Prokka, a command line software tool to fully annotate a draft bacterial genome in about 10 min on a typical desktop computer. It produces standards-compliant output files for further analysis or viewing in genome browsers. Prokka is implemented in Perl and is freely available under an open source GPLv2 license from http://vicbioinformatics.com/. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.
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              QUAST: quality assessment tool for genome assemblies.

              Limitations of genome sequencing techniques have led to dozens of assembly algorithms, none of which is perfect. A number of methods for comparing assemblers have been developed, but none is yet a recognized benchmark. Further, most existing methods for comparing assemblies are only applicable to new assemblies of finished genomes; the problem of evaluating assemblies of previously unsequenced species has not been adequately considered. Here, we present QUAST-a quality assessment tool for evaluating and comparing genome assemblies. This tool improves on leading assembly comparison software with new ideas and quality metrics. QUAST can evaluate assemblies both with a reference genome, as well as without a reference. QUAST produces many reports, summary tables and plots to help scientists in their research and in their publications. In this study, we used QUAST to compare several genome assemblers on three datasets. QUAST tables and plots for all of them are available in the Supplementary Material, and interactive versions of these reports are on the QUAST website. http://bioinf.spbau.ru/quast . Supplementary data are available at Bioinformatics online.
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                Author and article information

                Journal
                Sex Transm Infect
                Sex Transm Infect
                sextrans
                sti
                Sexually Transmitted Infections
                BMJ Publishing Group (BMA House, Tavistock Square, London, WC1H 9JR )
                1368-4973
                1472-3263
                February 2024
                9 November 2023
                : 100
                : 1
                : 25-30
                Affiliations
                [1 ] departmentExperimental Bacteriology , Ringgold_294567Institut Pasteur de Madagascar , Antananarivo, Madagascar
                [2 ] departmentCentre de Biologie Clinique , Ringgold_294567Institut Pasteur de Madagascar , Antananarivo, Madagascar
                [3 ] departmentCentre for Enteric Diseases , Ringgold_70687National Institute for Communicable Diseases , Johannesburg, South Africa
                [4 ] departmentDepartment of Medical Microbiology , Ringgold_56410University of Pretoria , Pretoria, South Africa
                Author notes
                [Correspondence to ] Dr Lala Fanomezantsoa Rafetrarivony, Experimental Bacteriology, Institut Pasteur de Madagascar, B.P. 1274, Ambatofotsikely Avaradoha 101 Antananarivo, Madagascar; fetra@ 123456pasteur.mg
                Author information
                http://orcid.org/0000-0003-1140-613X
                http://orcid.org/0000-0002-3637-7155
                http://orcid.org/0000-0002-2235-6038
                Article
                sextrans-2023-055878
                10.1136/sextrans-2023-055878
                10850657
                37945345
                e51ea00d-54d8-49f7-a31a-604b8594c325
                © Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

                This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.

                History
                : 26 May 2023
                : 08 October 2023
                Funding
                Funded by: Department of Health and Social Care’s Fleming Fund using UK aid;
                Award ID: N/A
                Categories
                Original Research
                1506
                Custom metadata
                unlocked

                Sexual medicine
                neisseria gonorrhoeae,africa,drug resistance, bacterial,molecular typing
                Sexual medicine
                neisseria gonorrhoeae, africa, drug resistance, bacterial, molecular typing

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