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      Identification of Antibiotic Resistance Genes in Orofacial Abscesses Using a Metagenomics-based Approach: A Pilot Study

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      https://www.riss.kr/link?id=A108650244

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      다국어 초록 (Multilingual Abstract)

      Purpose: Culture-based methods for microbiological diagnosis and antibiotic susceptibility tests have limitations in the management of orofacial infections. We aimed to profile pus microbiota and identify antibiotic resistance genes (ARGs) using a culture-independent approach.
      Materials and Methods: Genomic DNA samples extracted from the pus specimens of two patients with orofacial abscesses were subjected to shotgun sequencing on the NovaSeq system. Taxonomic profiling and prediction of ARGs were performed directly from the metagenomic raw reads.
      Result: Taxonomic profiling revealed obligate anaerobic polymicrobial communities associated with infections of odontogenic origins: the microbial community of Patient 1 consisted of one predominant species (Prevotella oris 74.6%) with 27 minor species, while the sample from Patient 2 contained 3 abundant species (Porphyromonas endodontalis 33.0%; P. oris 31.6%; and Prevotella koreensis 13.4%) with five minor species. A total of 150 and 136 putative ARGs were predicted in the metagenome of each pus sample. The coverage of most predicted ARGs was less than 10%, and only the CfxA2 gene identified in Patient 1 was covered 100%. ARG analysis of the seven assembled genome/metagenome datasets of P. oris revealed that strain C735 carried the CfxA2 gene.
      Conclusion: A metagenomics-based approach is useful to profile predominantly anaerobic polymicrobial communities but needs further verification for reliable ARG detection.
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      Purpose: Culture-based methods for microbiological diagnosis and antibiotic susceptibility tests have limitations in the management of orofacial infections. We aimed to profile pus microbiota and identify antibiotic resistance genes (ARGs) using a cul...

      Purpose: Culture-based methods for microbiological diagnosis and antibiotic susceptibility tests have limitations in the management of orofacial infections. We aimed to profile pus microbiota and identify antibiotic resistance genes (ARGs) using a culture-independent approach.
      Materials and Methods: Genomic DNA samples extracted from the pus specimens of two patients with orofacial abscesses were subjected to shotgun sequencing on the NovaSeq system. Taxonomic profiling and prediction of ARGs were performed directly from the metagenomic raw reads.
      Result: Taxonomic profiling revealed obligate anaerobic polymicrobial communities associated with infections of odontogenic origins: the microbial community of Patient 1 consisted of one predominant species (Prevotella oris 74.6%) with 27 minor species, while the sample from Patient 2 contained 3 abundant species (Porphyromonas endodontalis 33.0%; P. oris 31.6%; and Prevotella koreensis 13.4%) with five minor species. A total of 150 and 136 putative ARGs were predicted in the metagenome of each pus sample. The coverage of most predicted ARGs was less than 10%, and only the CfxA2 gene identified in Patient 1 was covered 100%. ARG analysis of the seven assembled genome/metagenome datasets of P. oris revealed that strain C735 carried the CfxA2 gene.
      Conclusion: A metagenomics-based approach is useful to profile predominantly anaerobic polymicrobial communities but needs further verification for reliable ARG detection.

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      참고문헌 (Reference)

      1 김이석 ; 박재홍 ; 전수성 ; 한동열 ; 김정은 ; 이병돈, "심경부 감염의 임상분석" 대한이비인후과학회 53 (53): 627-631, 2010

      2 Drawz SM, "Three decades of betalactamase inhibitors" 23 : 160-201, 2010

      3 Li H, "The sequence alignment/map format and SAMtools" 25 : 2078-2079, 2009

      4 Weise H, "Severe odontogenic infections with septic progress-a constant and increasing challenge : a retrospective analysis" 19 : 173-, 2019

      5 Rowe W, "Search engine for antimicrobial resistance : a cloud compatible pipeline and web interface for rapidly detecting antimicrobial resistance genes directly from sequence data" 10 : e0133492-, 2015

      6 De Angelis G, "Rapid molecular tests for detection of antimicrobial resistance determinants in Gram-negative organisms from positive blood cultures : a systematic review and meta-analysis" 26 : 271-280, 2020

      7 Clausen PTLC, "Rapid and precise alignment of raw reads against redundant databases with KMA" 19 : 307-, 2018

      8 Könönen E, "Prevotella species as oral residents and infectious agents with potential impact on systemic conditions" 14 : 2079814-, 2022

      9 Park SN, "Prevotella koreensis sp. nov., isolated from human subgingival dental plaque of periodontitis lesion" 76 : 1055-1060, 2019

      10 Rooney AM, "Performance characteristics of next-generation sequencing for the detection of antimicrobial resistance determinants in Escherichia coli genomes and metagenomes" 7 : e0002222-, 2022

      1 김이석 ; 박재홍 ; 전수성 ; 한동열 ; 김정은 ; 이병돈, "심경부 감염의 임상분석" 대한이비인후과학회 53 (53): 627-631, 2010

      2 Drawz SM, "Three decades of betalactamase inhibitors" 23 : 160-201, 2010

      3 Li H, "The sequence alignment/map format and SAMtools" 25 : 2078-2079, 2009

      4 Weise H, "Severe odontogenic infections with septic progress-a constant and increasing challenge : a retrospective analysis" 19 : 173-, 2019

      5 Rowe W, "Search engine for antimicrobial resistance : a cloud compatible pipeline and web interface for rapidly detecting antimicrobial resistance genes directly from sequence data" 10 : e0133492-, 2015

      6 De Angelis G, "Rapid molecular tests for detection of antimicrobial resistance determinants in Gram-negative organisms from positive blood cultures : a systematic review and meta-analysis" 26 : 271-280, 2020

      7 Clausen PTLC, "Rapid and precise alignment of raw reads against redundant databases with KMA" 19 : 307-, 2018

      8 Könönen E, "Prevotella species as oral residents and infectious agents with potential impact on systemic conditions" 14 : 2079814-, 2022

      9 Park SN, "Prevotella koreensis sp. nov., isolated from human subgingival dental plaque of periodontitis lesion" 76 : 1055-1060, 2019

      10 Rooney AM, "Performance characteristics of next-generation sequencing for the detection of antimicrobial resistance determinants in Escherichia coli genomes and metagenomes" 7 : e0002222-, 2022

      11 Warnke PH, "Penicillin compared with other advanced broad spectrum antibiotics regarding antibacterial activity against oral pathogens isolated from odontogenic abscesses" 36 : 462-467, 2008

      12 Kamiński B, "Oral and maxillofacial infections-a bacterial and clinical cross-section" 11 : 2731-, 2022

      13 Bahl R, "Odontogenic infections : microbiology and management" 5 : 307-311, 2014

      14 Beka D, "Microorganisms involved in deep neck infection(DNIs)in Greece : detection, identification and susceptibility to antimicrobials" 19 : 850-, 2019

      15 Siqueira JF Jr, "Microbiology and treatment of acute apical abscesses" 26 : 255-273, 2013

      16 Alauzet C, "Metronidazole resistance and nim genes in anaerobes : a review" 55 : 40-53, 2019

      17 Antelo V, "Metagenomic strategies identify diverse integron-integrase and antibiotic resistance genes in the Antarctic environment" 10 : e1219-, 2021

      18 Jevon P, "Management of odontogenic infections and sepsis : an update" 229 : 363-370, 2020

      19 Lakin SM, "MEGARes : an antimicrobial resistance database for high throughput sequencing" 45 : D574-80, 2017

      20 Yoon SH, "Introducing EzBioCloud : a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies" 67 : 1613-1617, 2017

      21 Chalita M, "Improved metagenomic taxonomic profiling using a curated core gene-based bacterial database reveals unrecognized species in the genus Streptococcus" 9 : 204-, 2020

      22 Wood DE, "Improved metagenomic analysis with Kraken 2" 20 : 257-, 2019

      23 Berglund F, "Identification and reconstruction of novel antibiotic resistance genes from metagenomes" 7 : 52-, 2019

      24 Lakin SM, "Hierarchical Hidden Markov models enable accurate and diverse detection of antimicrobial resistance sequences" 2 : 294-, 2019

      25 Mohd Asri NA, "Global prevalence of nosocomial multidrugresistant Klebsiella pneumoniae: a systematic review and meta-analysis" 10 : 1508-, 2021

      26 Leekha S, "General principles of antimicrobial therapy" 86 : 156-167, 2011

      27 Wybo I, "Fourth Belgian multicentre survey of antibiotic susceptibility of anaerobic bacteria" 69 : 155-161, 2014

      28 Langmead B, "Fast gapped-read alignment with Bowtie 2" 9 : 357-359, 2012

      29 Buchfink B, "Fast and sensitive protein alignment using DIAMOND" 12 : 59-60, 2015

      30 Hillmann B, "Evaluating the information content of shallow shotgun metagenomics" 3 : e00069-, 2018

      31 Horcajada JP, "Epidemiology and treatment of multidrug-resistant and extensively drug-resistant Pseudomonas aeruginosa infections" 32 : e00031-, 2019

      32 Leão-Vasconcelos LS, "Enterobacteriaceae isolates from the oral cavity of workers in a Brazilian oncology hospital" 57 : 121-127, 2015

      33 Colombo AP, "Comparisons of subgingival microbial profiles of refractory periodontitis, severe periodontitis, and periodontal health using the human oral microbe identification microarray" 80 : 1421-1432, 2009

      34 Böttger S, "Clinical relevance of the microbiome in odontogenic abscesses" 10 : 916-, 2021

      35 Alcock BP, "CARD 2020 : antibiotic resistome surveillance with the comprehensive antibiotic resistance database" 48 : D517-25, 2020

      36 Quinlan AR, "BEDTools : a flexible suite of utilities for comparing genomic features" 26 : 841-842, 2010

      37 Badr MT, "Antimicrobial susceptibility patterns and wild-type MIC distributions of anaerobic bacteria at a German University Hospital : a five-year retrospective study(2015-2019)" 9 : 823-, 2020

      38 Meinen A, "Antimicrobial resistance and the spectrum of pathogens in dental and oral-maxillofacial infections in hospitals and dental practices in Germany" 12 : 676108-, 2021

      39 Ramirez MS, "Amikacin : uses, resistance, and prospects for inhibition" 22 : 2267-, 2017

      40 Yang Y, "ARGs-OAP : online analysis pipeline for antibiotic resistance genes detection from metagenomic data using an integrated structured ARG-database" 32 : 2346-2351, 2016

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