Cargando…

Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry

The assessment of B-cell clonality is a critical component of the evaluation of suspected lymphoproliferative disorders, but analysis from formalin fixed paraffin embedded tissues can be challenging if fresh tissue is not available for flow cytometry. Immunohistochemical and conventional bright fiel...

Descripción completa

Detalles Bibliográficos
Autores principales: Guo, Ling, Wang, Zhen, Anderson, Courtney M., Doolittle, Emerald, Kernag, Siobhan, Cotta, Claudiu V., Ondrejka, Sarah L., Ma, Xiao-Jun, Cook, James R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5843495/
https://www.ncbi.nlm.nih.gov/pubmed/29052600
http://dx.doi.org/10.1038/modpathol.2017.142
_version_ 1783305094420234240
author Guo, Ling
Wang, Zhen
Anderson, Courtney M.
Doolittle, Emerald
Kernag, Siobhan
Cotta, Claudiu V.
Ondrejka, Sarah L.
Ma, Xiao-Jun
Cook, James R.
author_facet Guo, Ling
Wang, Zhen
Anderson, Courtney M.
Doolittle, Emerald
Kernag, Siobhan
Cotta, Claudiu V.
Ondrejka, Sarah L.
Ma, Xiao-Jun
Cook, James R.
author_sort Guo, Ling
collection PubMed
description The assessment of B-cell clonality is a critical component of the evaluation of suspected lymphoproliferative disorders, but analysis from formalin fixed paraffin embedded tissues can be challenging if fresh tissue is not available for flow cytometry. Immunohistochemical and conventional bright field in situ hybridization stains for kappa and lambda are effective for evaluation of plasma cells, but are often insufficiently sensitive to detect the much lower abundance of light chains present in B cells. We describe an ultrasensitive RNA in situ hybridization assay which has been adapted for use on an automated immunohistochemistry platform and compare results with flow cytometry in 203 consecutive tissues and 104 consecutive bone marrows. Overall, in 203 tissue biopsies, RNA in situ hybridization identified light chain restricted B-cells in 85 (42%) vs. 58 (29%) by flow cytometry. Within 83 B-cell non-Hodgkin lymphomas, RNA in situ hybridization identified a restricted B-cells in 74 (89%) vs. 56 (67%) by flow cytometry. B-cell clonality could be evaluated in only 23/104 (22%) bone marrow cases due to poor RNA preservation, but evaluable cases showed 91% concordance with flow cytometry. RNA in situ hybridization allowed for recognition of biclonal/composite lymphomas not identified by flow cytometry, and highlighted unexpected findings, such as coexpression of kappa and lambda RNA in 2 cases and the presence of lambda light chain RNA in a T lymphoblastic lymphoma. Automated RNA in situ hybridization showed excellent interobserver reproducibility for manual evaluation (average K=0.92), and an automated image analysis system showed high concordance (97%) with manual evaluation. Automated RNA in situ hybridization staining, which can be adopted on commonly utilized immunohistochemistry instruments, allows for the interpretation of clonality in the context of the morphologic features in formalin fixed, paraffin embedded tissues with a clinical sensitivity similar or superior to flow cytometry.
format Online
Article
Text
id pubmed-5843495
institution National Center for Biotechnology Information
language English
publishDate 2017
record_format MEDLINE/PubMed
spelling pubmed-58434952018-04-20 Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry Guo, Ling Wang, Zhen Anderson, Courtney M. Doolittle, Emerald Kernag, Siobhan Cotta, Claudiu V. Ondrejka, Sarah L. Ma, Xiao-Jun Cook, James R. Mod Pathol Article The assessment of B-cell clonality is a critical component of the evaluation of suspected lymphoproliferative disorders, but analysis from formalin fixed paraffin embedded tissues can be challenging if fresh tissue is not available for flow cytometry. Immunohistochemical and conventional bright field in situ hybridization stains for kappa and lambda are effective for evaluation of plasma cells, but are often insufficiently sensitive to detect the much lower abundance of light chains present in B cells. We describe an ultrasensitive RNA in situ hybridization assay which has been adapted for use on an automated immunohistochemistry platform and compare results with flow cytometry in 203 consecutive tissues and 104 consecutive bone marrows. Overall, in 203 tissue biopsies, RNA in situ hybridization identified light chain restricted B-cells in 85 (42%) vs. 58 (29%) by flow cytometry. Within 83 B-cell non-Hodgkin lymphomas, RNA in situ hybridization identified a restricted B-cells in 74 (89%) vs. 56 (67%) by flow cytometry. B-cell clonality could be evaluated in only 23/104 (22%) bone marrow cases due to poor RNA preservation, but evaluable cases showed 91% concordance with flow cytometry. RNA in situ hybridization allowed for recognition of biclonal/composite lymphomas not identified by flow cytometry, and highlighted unexpected findings, such as coexpression of kappa and lambda RNA in 2 cases and the presence of lambda light chain RNA in a T lymphoblastic lymphoma. Automated RNA in situ hybridization showed excellent interobserver reproducibility for manual evaluation (average K=0.92), and an automated image analysis system showed high concordance (97%) with manual evaluation. Automated RNA in situ hybridization staining, which can be adopted on commonly utilized immunohistochemistry instruments, allows for the interpretation of clonality in the context of the morphologic features in formalin fixed, paraffin embedded tissues with a clinical sensitivity similar or superior to flow cytometry. 2017-10-20 2018-03 /pmc/articles/PMC5843495/ /pubmed/29052600 http://dx.doi.org/10.1038/modpathol.2017.142 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Guo, Ling
Wang, Zhen
Anderson, Courtney M.
Doolittle, Emerald
Kernag, Siobhan
Cotta, Claudiu V.
Ondrejka, Sarah L.
Ma, Xiao-Jun
Cook, James R.
Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry
title Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry
title_full Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry
title_fullStr Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry
title_full_unstemmed Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry
title_short Ultrasensitive Automated RNA in situ Hybridization for Kappa and Lambda Light Chain mRNA Detects B-cell Clonality in Tissue Biopsies with Performance Comparable or Superior to Flow Cytometry
title_sort ultrasensitive automated rna in situ hybridization for kappa and lambda light chain mrna detects b-cell clonality in tissue biopsies with performance comparable or superior to flow cytometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5843495/
https://www.ncbi.nlm.nih.gov/pubmed/29052600
http://dx.doi.org/10.1038/modpathol.2017.142
work_keys_str_mv AT guoling ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT wangzhen ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT andersoncourtneym ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT doolittleemerald ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT kernagsiobhan ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT cottaclaudiuv ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT ondrejkasarahl ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT maxiaojun ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry
AT cookjamesr ultrasensitiveautomatedrnainsituhybridizationforkappaandlambdalightchainmrnadetectsbcellclonalityintissuebiopsieswithperformancecomparableorsuperiortoflowcytometry