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2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori

BACKGROUND: Helicobacter pylori is associated with peptic ulcer disease and gastric malignancy. Antimicrobial susceptibility testing (AST) is often requested for patients who fail eradication therapy. The CLSI reference method, agar dilution (AD), is not performed in most laboratories and maintainin...

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Autores principales: Shakir, Salika M, Otiso, Joshua, Keller, George, Heule, Hillary Van, Kohner, Peggy, Cole, Nicolynn, Schuetz, Audrey N, Richter, Sandra S, Couturier, Marc Roger
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6254197/
http://dx.doi.org/10.1093/ofid/ofy210.1728
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author Shakir, Salika M
Otiso, Joshua
Keller, George
Heule, Hillary Van
Kohner, Peggy
Cole, Nicolynn
Schuetz, Audrey N
Richter, Sandra S
Couturier, Marc Roger
author_facet Shakir, Salika M
Otiso, Joshua
Keller, George
Heule, Hillary Van
Kohner, Peggy
Cole, Nicolynn
Schuetz, Audrey N
Richter, Sandra S
Couturier, Marc Roger
author_sort Shakir, Salika M
collection PubMed
description BACKGROUND: Helicobacter pylori is associated with peptic ulcer disease and gastric malignancy. Antimicrobial susceptibility testing (AST) is often requested for patients who fail eradication therapy. The CLSI reference method, agar dilution (AD), is not performed in most laboratories and maintaining organism viability during transit to a reference laboratory is difficult. We assessed the performance of the Etest (bioMerieux) as a method for H. pylori AST in comparison to AD. METHODS: Frozen stocks of 82 H. pylori isolates with AD results previously reported by Mayo Clinic were prepared from the same plate for distribution to participating laboratories. Etest was performed at ARUP Laboratories and Cleveland Clinic (CC). For Etest, isolates were incubated for 72 hours in a microaerobic atmosphere. Aged Mueller–Hinton agar with 5% sheep blood plates were inoculated with a three McFarland suspension prepared in brain heart infusion broth. Etest strips were applied and MICs read after 72 hours of microaerobic incubation. Results were interpreted by applying CLSI and EUCAST breakpoints. Categorical agreement (CA), very major, major and minor errors (VME, ME, and mE) were determined for Etest using AD as the reference method. Isolates with errors were repeat tested in duplicate by Etest to determine the final results summarized below. RESULTS: For clarithromycin, 65% of isolates were resistant (R) by AD; Etest results at each laboratory showed 97.5% CA (1 mE and 1 ME). For tetracycline, only 2.5% of isolates were R by AD; a single VME occurred at both ARUP and CC (98.8% CA) with the same isolate. The AD dilutions tested for amoxicillin prevented interpretation with EUCAST breakpoints. With one exception, amoxicillin Etest results were susceptible (S, ≤0.125 mg/L) at both laboratories (98.8% of MICs ± one dilution). Applying levofloxacin EUCAST breakpoints (S, ≤1 mg/L) to interpret ciprofloxacin results, 57.8% of isolates were R by AD. ARUP CA was 97.5% (1 ME, 1 VME) and CC CA was 96.3% (1 ME, 2 VMEs). CONCLUSION: Clarithromycin, tetracycline, and ciprofloxacin Etest results for H. pylori showed acceptable CA (>95%) at both testing sites compared with the AD reference method. The comparative ease of performance and reproducibility of the Etest may help standardize it as an AST method for H. pylori. DISCLOSURES: S. S. Richter, bioMerieux: Grant Investigator, Research grant; BD Diagnostics: Grant Investigator, Research grant; Roche: Grant Investigator, Research grant; Hologic: Grant Investigator, Research grant; Diasorin: Grant Investigator, Research grant; Accelerate: Grant Investigator, Research grant; Biofire: Grant Investigator, Research grant.
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spelling pubmed-62541972018-11-28 2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori Shakir, Salika M Otiso, Joshua Keller, George Heule, Hillary Van Kohner, Peggy Cole, Nicolynn Schuetz, Audrey N Richter, Sandra S Couturier, Marc Roger Open Forum Infect Dis Abstracts BACKGROUND: Helicobacter pylori is associated with peptic ulcer disease and gastric malignancy. Antimicrobial susceptibility testing (AST) is often requested for patients who fail eradication therapy. The CLSI reference method, agar dilution (AD), is not performed in most laboratories and maintaining organism viability during transit to a reference laboratory is difficult. We assessed the performance of the Etest (bioMerieux) as a method for H. pylori AST in comparison to AD. METHODS: Frozen stocks of 82 H. pylori isolates with AD results previously reported by Mayo Clinic were prepared from the same plate for distribution to participating laboratories. Etest was performed at ARUP Laboratories and Cleveland Clinic (CC). For Etest, isolates were incubated for 72 hours in a microaerobic atmosphere. Aged Mueller–Hinton agar with 5% sheep blood plates were inoculated with a three McFarland suspension prepared in brain heart infusion broth. Etest strips were applied and MICs read after 72 hours of microaerobic incubation. Results were interpreted by applying CLSI and EUCAST breakpoints. Categorical agreement (CA), very major, major and minor errors (VME, ME, and mE) were determined for Etest using AD as the reference method. Isolates with errors were repeat tested in duplicate by Etest to determine the final results summarized below. RESULTS: For clarithromycin, 65% of isolates were resistant (R) by AD; Etest results at each laboratory showed 97.5% CA (1 mE and 1 ME). For tetracycline, only 2.5% of isolates were R by AD; a single VME occurred at both ARUP and CC (98.8% CA) with the same isolate. The AD dilutions tested for amoxicillin prevented interpretation with EUCAST breakpoints. With one exception, amoxicillin Etest results were susceptible (S, ≤0.125 mg/L) at both laboratories (98.8% of MICs ± one dilution). Applying levofloxacin EUCAST breakpoints (S, ≤1 mg/L) to interpret ciprofloxacin results, 57.8% of isolates were R by AD. ARUP CA was 97.5% (1 ME, 1 VME) and CC CA was 96.3% (1 ME, 2 VMEs). CONCLUSION: Clarithromycin, tetracycline, and ciprofloxacin Etest results for H. pylori showed acceptable CA (>95%) at both testing sites compared with the AD reference method. The comparative ease of performance and reproducibility of the Etest may help standardize it as an AST method for H. pylori. DISCLOSURES: S. S. Richter, bioMerieux: Grant Investigator, Research grant; BD Diagnostics: Grant Investigator, Research grant; Roche: Grant Investigator, Research grant; Hologic: Grant Investigator, Research grant; Diasorin: Grant Investigator, Research grant; Accelerate: Grant Investigator, Research grant; Biofire: Grant Investigator, Research grant. Oxford University Press 2018-11-26 /pmc/articles/PMC6254197/ http://dx.doi.org/10.1093/ofid/ofy210.1728 Text en © The Author(s) 2018. 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 Abstracts
Shakir, Salika M
Otiso, Joshua
Keller, George
Heule, Hillary Van
Kohner, Peggy
Cole, Nicolynn
Schuetz, Audrey N
Richter, Sandra S
Couturier, Marc Roger
2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori
title 2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori
title_full 2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori
title_fullStr 2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori
title_full_unstemmed 2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori
title_short 2072. Multicenter Evaluation of the Etest vs. Agar Dilution for Susceptibility Testing of Helicobacter pylori
title_sort 2072. multicenter evaluation of the etest vs. agar dilution for susceptibility testing of helicobacter pylori
topic Abstracts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6254197/
http://dx.doi.org/10.1093/ofid/ofy210.1728
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