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Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET

Diagnostic assays for molecular alterations highly correlated with prognosis, predictive efficacy or safety of therapeutics are valuable clinical tools and in some cases approved as companion diagnostics (CDx) by the Federal Food and Drug Administration. For example, assays that determine echinoderm...

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Autores principales: Miller, Rachel, Thorne-Nuzzo, Trish, Loftin, Isabell, McElhinny, Abigail, Towne, Penny, Clements, June
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Lippincott Williams & Wilkins 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7253184/
https://www.ncbi.nlm.nih.gov/pubmed/30724750
http://dx.doi.org/10.1097/PAI.0000000000000730
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author Miller, Rachel
Thorne-Nuzzo, Trish
Loftin, Isabell
McElhinny, Abigail
Towne, Penny
Clements, June
author_facet Miller, Rachel
Thorne-Nuzzo, Trish
Loftin, Isabell
McElhinny, Abigail
Towne, Penny
Clements, June
author_sort Miller, Rachel
collection PubMed
description Diagnostic assays for molecular alterations highly correlated with prognosis, predictive efficacy or safety of therapeutics are valuable clinical tools and in some cases approved as companion diagnostics (CDx) by the Federal Food and Drug Administration. For example, assays that determine echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) translocation status have been approved as CDx assay for therapies that target this molecular alteration. Characterizing the parameters that may compromise diagnostic accuracy for molecular biomarkers is critical for optimal patient care. To investigate the impact of pre-analytical handling and processing of tumor tissue on commonly used diagnostic immunohistochemistry-based assays for ALK and mesenchymal epithelial transition protein [c-mesenchymal epithelial transition (c-MET)], we investigated the effects of cold ischemia, fixative type, fixation time, and cut-slide age on staining consistency and intensity using human lung xenograft tumor tissue. Cold ischemia times for up to 5 to 6 hours for c-MET or ALK, respectively had minimal impact on staining. The optimal fixation conditions for both assays were found to be at least 6 hours and up to 48 hours for c-MET or 72 hours for ALK, in 10% neutral buffered formalin and Zinc formalin. The ALK antigen demonstrated marked staining intensity differences across non-neutral buffered formalin fixative types and times. Finally, cut-slide age influenced assay performance for both ALK and c-MET, with maximum stability observed when cut slides were stored at ambient temperatures (30°C) for no longer than 3, and 5 months, respectively. This study highlights the potential for pre-analytical factors to confound diagnostic test result interpretation.
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spelling pubmed-72531842020-06-29 Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET Miller, Rachel Thorne-Nuzzo, Trish Loftin, Isabell McElhinny, Abigail Towne, Penny Clements, June Appl Immunohistochem Mol Morphol Research Articles Diagnostic assays for molecular alterations highly correlated with prognosis, predictive efficacy or safety of therapeutics are valuable clinical tools and in some cases approved as companion diagnostics (CDx) by the Federal Food and Drug Administration. For example, assays that determine echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) translocation status have been approved as CDx assay for therapies that target this molecular alteration. Characterizing the parameters that may compromise diagnostic accuracy for molecular biomarkers is critical for optimal patient care. To investigate the impact of pre-analytical handling and processing of tumor tissue on commonly used diagnostic immunohistochemistry-based assays for ALK and mesenchymal epithelial transition protein [c-mesenchymal epithelial transition (c-MET)], we investigated the effects of cold ischemia, fixative type, fixation time, and cut-slide age on staining consistency and intensity using human lung xenograft tumor tissue. Cold ischemia times for up to 5 to 6 hours for c-MET or ALK, respectively had minimal impact on staining. The optimal fixation conditions for both assays were found to be at least 6 hours and up to 48 hours for c-MET or 72 hours for ALK, in 10% neutral buffered formalin and Zinc formalin. The ALK antigen demonstrated marked staining intensity differences across non-neutral buffered formalin fixative types and times. Finally, cut-slide age influenced assay performance for both ALK and c-MET, with maximum stability observed when cut slides were stored at ambient temperatures (30°C) for no longer than 3, and 5 months, respectively. This study highlights the potential for pre-analytical factors to confound diagnostic test result interpretation. Lippincott Williams & Wilkins 2020 2019-04-01 /pmc/articles/PMC7253184/ /pubmed/30724750 http://dx.doi.org/10.1097/PAI.0000000000000730 Text en Copyright © 2019 The Author(s). Published by Wolters Kluwer Health, Inc. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/) (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal. http://creativecommons.org/licenses/by-nc-nd/4.0/
spellingShingle Research Articles
Miller, Rachel
Thorne-Nuzzo, Trish
Loftin, Isabell
McElhinny, Abigail
Towne, Penny
Clements, June
Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET
title Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET
title_full Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET
title_fullStr Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET
title_full_unstemmed Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET
title_short Impact of Pre-Analytical Conditions on the Antigenicity of Lung Markers: ALK and MET
title_sort impact of pre-analytical conditions on the antigenicity of lung markers: alk and met
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7253184/
https://www.ncbi.nlm.nih.gov/pubmed/30724750
http://dx.doi.org/10.1097/PAI.0000000000000730
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