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826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods

BACKGROUND: Epidemiological (EPI) analyses of bacterial pathogens play an important role in infection control practices during suspected outbreaks. Pulsed field gel electrophoresis (PFGE) is the gold standard for EPI typing of most bacterial organisms, but this method has been slowly replaced by seq...

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Autores principales: Deshpande, Lalitagauri M, Collingsworth, Timothy, Mendes, Rodrigo E, Castanheira, Mariana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7777629/
http://dx.doi.org/10.1093/ofid/ofaa439.1015
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author Deshpande, Lalitagauri M
Collingsworth, Timothy
Mendes, Rodrigo E
Castanheira, Mariana
author_facet Deshpande, Lalitagauri M
Collingsworth, Timothy
Mendes, Rodrigo E
Castanheira, Mariana
author_sort Deshpande, Lalitagauri M
collection PubMed
description BACKGROUND: Epidemiological (EPI) analyses of bacterial pathogens play an important role in infection control practices during suspected outbreaks. Pulsed field gel electrophoresis (PFGE) is the gold standard for EPI typing of most bacterial organisms, but this method has been slowly replaced by sequencing-based methods, such as multilocus sequence typing (MLST) and core genome (cg)MLST that uses whole genome sequencing (WGS) data. We evaluated the utility of WGS ad hoc schemas to predict relatedness among Serratia marcescens (SM) clinical strains. METHODS: A total of 19 SM clinical isolates collected as part of the SENTRY Program and JMI worldwide collections were selected. Isolates were typed by PFGE and analyzed using GelCompar II (100% similarity scored as identical, > 85% and < 100% as genetically related and < 85% as unrelated). WGS was performed using MiSeq (Illumina) and contigs generated using SPAdes. Raw reads and assembled contigs were used to generate phylogenetic trees using kWIP and progressiveMauve (pMauve), respectively. Similarity matrices and dendrograms created by the three protocols were compared. RESULTS: Based on PFGE analysis, 19 isolates were classified into 10 pulsotypes, A through J, and 1 subtype, A1 (Fig.1A). Among dendrograms generated based on WGS, analysis of short k-mers (Fig.1B) inaccurately showed phylogenetic separation among isolates in A/A1, C and G types, while analysis of much longer contigs (Fig.1C) accurately clustered isolates according to groups defined using PFGE. One isolate in the A/A1 group (517323) was separated from the group using kWIP; however, using the more sophisticated pMauve, this isolate was accurately clustered within the A/A1 isolates. CONCLUSION: Concordance was observed between PFGE- and WGS-based phylogenetic analysis of SM for genetic relatedness. Based on this analysis, WGS data can be used to predict EPI of this bacterial species on an ad hoc basis. This methodology can be expanded to other species. Figure 1 [Image: see text] DISCLOSURES: Rodrigo E. Mendes, PhD, A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Allergan (Research Grant or Support)Allergan (Research Grant or Support)Basilea Pharmaceutica International, Ltd (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Department of Health and Human Services (Research Grant or Support)GlaxoSmithKline (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Merck (Research Grant or Support)Merck (Research Grant or Support)Pfizer (Research Grant or Support) Mariana Castanheira, PhD, 1928 Diagnostics (Research Grant or Support)A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Allergan (Research Grant or Support)Allergan (Research Grant or Support)Amplyx Pharmaceuticals (Research Grant or Support)Cidara Therapeutics (Research Grant or Support)Cidara Therapeutics (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Fox Chase Chemical Diversity Center (Research Grant or Support)GlaxoSmithKline (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Merck (Research Grant or Support)Merck (Research Grant or Support)Merck & Co, Inc. (Research Grant or Support)Merck & Co, Inc. (Research Grant or Support)Paratek Pharma, LLC (Research Grant or Support)Pfizer (Research Grant or Support)Qpex Biopharma (Research Grant or Support)
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spelling pubmed-77776292021-01-07 826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods Deshpande, Lalitagauri M Collingsworth, Timothy Mendes, Rodrigo E Castanheira, Mariana Open Forum Infect Dis Poster Abstracts BACKGROUND: Epidemiological (EPI) analyses of bacterial pathogens play an important role in infection control practices during suspected outbreaks. Pulsed field gel electrophoresis (PFGE) is the gold standard for EPI typing of most bacterial organisms, but this method has been slowly replaced by sequencing-based methods, such as multilocus sequence typing (MLST) and core genome (cg)MLST that uses whole genome sequencing (WGS) data. We evaluated the utility of WGS ad hoc schemas to predict relatedness among Serratia marcescens (SM) clinical strains. METHODS: A total of 19 SM clinical isolates collected as part of the SENTRY Program and JMI worldwide collections were selected. Isolates were typed by PFGE and analyzed using GelCompar II (100% similarity scored as identical, > 85% and < 100% as genetically related and < 85% as unrelated). WGS was performed using MiSeq (Illumina) and contigs generated using SPAdes. Raw reads and assembled contigs were used to generate phylogenetic trees using kWIP and progressiveMauve (pMauve), respectively. Similarity matrices and dendrograms created by the three protocols were compared. RESULTS: Based on PFGE analysis, 19 isolates were classified into 10 pulsotypes, A through J, and 1 subtype, A1 (Fig.1A). Among dendrograms generated based on WGS, analysis of short k-mers (Fig.1B) inaccurately showed phylogenetic separation among isolates in A/A1, C and G types, while analysis of much longer contigs (Fig.1C) accurately clustered isolates according to groups defined using PFGE. One isolate in the A/A1 group (517323) was separated from the group using kWIP; however, using the more sophisticated pMauve, this isolate was accurately clustered within the A/A1 isolates. CONCLUSION: Concordance was observed between PFGE- and WGS-based phylogenetic analysis of SM for genetic relatedness. Based on this analysis, WGS data can be used to predict EPI of this bacterial species on an ad hoc basis. This methodology can be expanded to other species. Figure 1 [Image: see text] DISCLOSURES: Rodrigo E. Mendes, PhD, A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Allergan (Research Grant or Support)Allergan (Research Grant or Support)Basilea Pharmaceutica International, Ltd (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Department of Health and Human Services (Research Grant or Support)GlaxoSmithKline (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Merck (Research Grant or Support)Merck (Research Grant or Support)Pfizer (Research Grant or Support) Mariana Castanheira, PhD, 1928 Diagnostics (Research Grant or Support)A. Menarini Industrie Farmaceutiche Riunite S.R.L. (Research Grant or Support)Allergan (Research Grant or Support)Allergan (Research Grant or Support)Amplyx Pharmaceuticals (Research Grant or Support)Cidara Therapeutics (Research Grant or Support)Cidara Therapeutics (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Cipla Ltd. (Research Grant or Support)Fox Chase Chemical Diversity Center (Research Grant or Support)GlaxoSmithKline (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Merck (Research Grant or Support)Merck (Research Grant or Support)Merck & Co, Inc. (Research Grant or Support)Merck & Co, Inc. (Research Grant or Support)Paratek Pharma, LLC (Research Grant or Support)Pfizer (Research Grant or Support)Qpex Biopharma (Research Grant or Support) Oxford University Press 2020-12-31 /pmc/articles/PMC7777629/ http://dx.doi.org/10.1093/ofid/ofaa439.1015 Text en © The Author 2020. Published by Oxford University Press on behalf of Infectious Diseases Society of America. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Poster Abstracts
Deshpande, Lalitagauri M
Collingsworth, Timothy
Mendes, Rodrigo E
Castanheira, Mariana
826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods
title 826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods
title_full 826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods
title_fullStr 826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods
title_full_unstemmed 826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods
title_short 826. Comparative Epidemiological Analysis of Serratia marcescens using PFGE and Whole Genome Sequence Methods
title_sort 826. comparative epidemiological analysis of serratia marcescens using pfge and whole genome sequence methods
topic Poster Abstracts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7777629/
http://dx.doi.org/10.1093/ofid/ofaa439.1015
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