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2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?

BACKGROUND: As part of active surveillance in our NICU for methicillin-resistant Staphylococcus aureus (MRSA), two isolates representing modified S. aureus (MODSA), which are Methicillin resistant but lacking mecA or C were identified. Our current microbiology laboratory workflow for screening for M...

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Autores principales: Gitman, Melissa R, Obla, Ajay, Van de Guchte, Adriana, Sordillo, Emilia Mia, Polanco, Jose, Chung, Marilyn, Oussenko, Irina, Smith, Melissa L, Altman, Deena R, Van Bakel, Harm
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6810657/
http://dx.doi.org/10.1093/ofid/ofz360.1809
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author Gitman, Melissa R
Obla, Ajay
Van de Guchte, Adriana
Sordillo, Emilia Mia
Polanco, Jose
Chung, Marilyn
Oussenko, Irina
Smith, Melissa L
Altman, Deena R
Van Bakel, Harm
author_facet Gitman, Melissa R
Obla, Ajay
Van de Guchte, Adriana
Sordillo, Emilia Mia
Polanco, Jose
Chung, Marilyn
Oussenko, Irina
Smith, Melissa L
Altman, Deena R
Van Bakel, Harm
author_sort Gitman, Melissa R
collection PubMed
description BACKGROUND: As part of active surveillance in our NICU for methicillin-resistant Staphylococcus aureus (MRSA), two isolates representing modified S. aureus (MODSA), which are Methicillin resistant but lacking mecA or C were identified. Our current microbiology laboratory workflow for screening for MRSA involves plating isolates on chromoID agar (bioMérieux, Marcy-l’Etoile, France) as well as on sheep blood agar (SBA). Β hemolytic colonies on SBA that are catalase and coagulase positive are set up for confirmation and antimicrobial susceptibility testing on the Vitek 2 (bioMérieux, Marcy-l’Etoile, France). METHODS: These 2 isolates (from Baby 1 and Baby 2) tested positive for green colonies on the chromoagar plates. The Vitek 2 subsequently identified both these isolates as MRSA. However, for research purposes, all positive NICU MRSA isolates are tested via whole-genome sequencing (WGS). Both isolates were identified by WGS as methicillin-susceptible Staphylococcus aureus (MSSA). We subsequently went back and performed additional workup on these isolates. Isolates were plated on SBA and chromagar again and incubated for 24 hours. 2 colonies of different morphologies from the chromagar plates and 3 from the SBA were randomly selected and subcultured to chromagar and SBA for a total of 5 subcultures. Each of the subcultures was tested using staphaurex, mannitol salt agar and the Cepheid Xpert MRSA assay and all were confirmed to be Staphylococcus aureus. RESULTS: All 10 isolates tested negative by the Cepheid Xpert MRSA assay for MRSA. Phenotypic testing was set up again for all ten isolates using the vitek GP panel, as well as cefoxitin disk, and oxacillin E test using Mueller–Hinton agar supplemented with 2% NaCl as per CLSI methods. See table attached for results. CONCLUSION: In conclusion, these two cases highlight the difficulty in identifying non-MecA, non-MecC-mediated MRSA isolates in the clinical microbiology laboratory. This is particularly important as more laboratories rely on testing for MecA by PCR for surveillance testing. These 2 cases were further complicated by heterogeneous sub-populations of Staphylococcus aureus. Failure to recognize these variant forms of MRSA can lead to difficulties in implementing appropriate therapy and infection control measures. Improved methodologies are needed. [Image: see text] DISCLOSURES: All authors: No reported disclosures.
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spelling pubmed-68106572019-10-28 2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus? Gitman, Melissa R Obla, Ajay Van de Guchte, Adriana Sordillo, Emilia Mia Polanco, Jose Chung, Marilyn Oussenko, Irina Smith, Melissa L Altman, Deena R Van Bakel, Harm Open Forum Infect Dis Abstracts BACKGROUND: As part of active surveillance in our NICU for methicillin-resistant Staphylococcus aureus (MRSA), two isolates representing modified S. aureus (MODSA), which are Methicillin resistant but lacking mecA or C were identified. Our current microbiology laboratory workflow for screening for MRSA involves plating isolates on chromoID agar (bioMérieux, Marcy-l’Etoile, France) as well as on sheep blood agar (SBA). Β hemolytic colonies on SBA that are catalase and coagulase positive are set up for confirmation and antimicrobial susceptibility testing on the Vitek 2 (bioMérieux, Marcy-l’Etoile, France). METHODS: These 2 isolates (from Baby 1 and Baby 2) tested positive for green colonies on the chromoagar plates. The Vitek 2 subsequently identified both these isolates as MRSA. However, for research purposes, all positive NICU MRSA isolates are tested via whole-genome sequencing (WGS). Both isolates were identified by WGS as methicillin-susceptible Staphylococcus aureus (MSSA). We subsequently went back and performed additional workup on these isolates. Isolates were plated on SBA and chromagar again and incubated for 24 hours. 2 colonies of different morphologies from the chromagar plates and 3 from the SBA were randomly selected and subcultured to chromagar and SBA for a total of 5 subcultures. Each of the subcultures was tested using staphaurex, mannitol salt agar and the Cepheid Xpert MRSA assay and all were confirmed to be Staphylococcus aureus. RESULTS: All 10 isolates tested negative by the Cepheid Xpert MRSA assay for MRSA. Phenotypic testing was set up again for all ten isolates using the vitek GP panel, as well as cefoxitin disk, and oxacillin E test using Mueller–Hinton agar supplemented with 2% NaCl as per CLSI methods. See table attached for results. CONCLUSION: In conclusion, these two cases highlight the difficulty in identifying non-MecA, non-MecC-mediated MRSA isolates in the clinical microbiology laboratory. This is particularly important as more laboratories rely on testing for MecA by PCR for surveillance testing. These 2 cases were further complicated by heterogeneous sub-populations of Staphylococcus aureus. Failure to recognize these variant forms of MRSA can lead to difficulties in implementing appropriate therapy and infection control measures. Improved methodologies are needed. [Image: see text] DISCLOSURES: All authors: No reported disclosures. Oxford University Press 2019-10-23 /pmc/articles/PMC6810657/ http://dx.doi.org/10.1093/ofid/ofz360.1809 Text en © The Author(s) 2019. 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
Gitman, Melissa R
Obla, Ajay
Van de Guchte, Adriana
Sordillo, Emilia Mia
Polanco, Jose
Chung, Marilyn
Oussenko, Irina
Smith, Melissa L
Altman, Deena R
Van Bakel, Harm
2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?
title 2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?
title_full 2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?
title_fullStr 2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?
title_full_unstemmed 2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?
title_short 2129. When Is Methicillin-resistant Staphylococcus aureus not Methicillin-resistant Staphylococcus aureus?
title_sort 2129. when is methicillin-resistant staphylococcus aureus not methicillin-resistant staphylococcus aureus?
topic Abstracts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6810657/
http://dx.doi.org/10.1093/ofid/ofz360.1809
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