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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Lippincott Williams & Wilkins
2020
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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. |
format | Online Article Text |
id | pubmed-7253184 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Lippincott Williams & Wilkins |
record_format | MEDLINE/PubMed |
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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