A1 Refereed original research article in a scientific journal
Use of antisera directed against dsRNA to detect viral infections in formalin-fixed paraffin-embedded tissue
Authors: Richardson SJ, Willcox A, Hilton DA, Tauriainen S, Hyoty H, Bone AJ, Foulis AK, Morgan NG
Publisher: ELSEVIER SCIENCE BV
Publication year: 2010
Journal: Journal of Clinical Virology
Journal name in source: JOURNAL OF CLINICAL VIROLOGY
Journal acronym: J CLIN VIROL
Volume: 49
Issue: 3
First page : 180
Last page: 185
Number of pages: 6
ISSN: 1386-6532
DOI: https://doi.org/10.1016/j.jcv.2010.07.015
Abstract
Background: The detection of viral infection in paraffin-embedded, formalin-fixed tissue is notoriously
difficult and often requires inherent knowledge about the specific virus being sought. For this reason,
there is an ongoing need for reagents and methods which can identify a range of different virus types in
paraffin embedded tissue.
Objectives: The aim of this study was to optimise and validate the use of antisera directed against dsRNA
(>50 bp in length) in paraffin-embedded formalin-fixed tissue samples.
Study design: dsRNA antisera were optimised for use in a range of virally-infected tissue culture cells,
Coxsackie-infected mice and human tissues. The specificity of labelling was confirmed by pre-adsorption
of antisera with poly-IC and by digestion of dsRNA with RNaseIII.
Results: Two different polyclonal dsRNA antisera (J2 and K1) were capable of recognising dsRNA encoded
by all the multiple different viral types (including (+) ssRNA viruses, dsRNA viruses and DNA viruses)
tested in paraffin-embedded formalin fixed infected cells and tissues. In contrast, the enteroviral vp1
antisera detected only a subset of the (+) ssRNA viruses tested. Staining was not seen in uninfected
cells or in uninfected control tissues. Positive staining was ablated following incubation of antisera with
poly-IC or by pre-treating sections with RNaseIII prior to staining.
Conclusions: The dsRNA antisera J2 and K1 are useful for the detection of viral infection in formalin-fixed,
paraffin-embedded, human tissue samples.. (C) 2010 Elsevier B. V. All rights reserved.
Background: The detection of viral infection in paraffin-embedded, formalin-fixed tissue is notoriously
difficult and often requires inherent knowledge about the specific virus being sought. For this reason,
there is an ongoing need for reagents and methods which can identify a range of different virus types in
paraffin embedded tissue.
Objectives: The aim of this study was to optimise and validate the use of antisera directed against dsRNA
(>50 bp in length) in paraffin-embedded formalin-fixed tissue samples.
Study design: dsRNA antisera were optimised for use in a range of virally-infected tissue culture cells,
Coxsackie-infected mice and human tissues. The specificity of labelling was confirmed by pre-adsorption
of antisera with poly-IC and by digestion of dsRNA with RNaseIII.
Results: Two different polyclonal dsRNA antisera (J2 and K1) were capable of recognising dsRNA encoded
by all the multiple different viral types (including (+) ssRNA viruses, dsRNA viruses and DNA viruses)
tested in paraffin-embedded formalin fixed infected cells and tissues. In contrast, the enteroviral vp1
antisera detected only a subset of the (+) ssRNA viruses tested. Staining was not seen in uninfected
cells or in uninfected control tissues. Positive staining was ablated following incubation of antisera with
poly-IC or by pre-treating sections with RNaseIII prior to staining.
Conclusions: The dsRNA antisera J2 and K1 are useful for the detection of viral infection in formalin-fixed,
paraffin-embedded, human tissue samples.. (C) 2010 Elsevier B. V. All rights reserved.
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