Immunohistochemical detection of double‐stranded RNA in formalin‐fixed paraffin‐embedded tissue

Double‐stranded RNA (dsRNA) is produced during most viral infections, and immunohistochemical detection of dsRNA has been proposed as a potential screening marker for viral replication. The anti‐dsRNA monoclonal antibody clone 9D5 is more sensitive than the established clone J2 but has not been vali...

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Published in:APMIS : acta pathologica, microbiologica et immunologica Scandinavica microbiologica et immunologica Scandinavica, 2023-05, Vol.131 (5), p.197-205
Main Authors: Thomsen, Christian, Røge, Rasmus, Fred, Åsa, Wanders, Alkwin
Format: Article
Language:English
Subjects:
COVID-19
Double-stranded RNA
Epitopes
FFPE
Formaldehyde
formalin
Humans
Immunohistochemistry
Infections
Monoclonal antibodies
Paraffin
Paraffin Embedding
Paraffins
Ribonuclease III
RNA, Double-Stranded
SARS-CoV-2
Severe acute respiratory syndrome coronavirus 2
Staining
Tissues
Viral diseases
Viral infections
virus
Virus Diseases
Viruses
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Summary:Double‐stranded RNA (dsRNA) is produced during most viral infections, and immunohistochemical detection of dsRNA has been proposed as a potential screening marker for viral replication. The anti‐dsRNA monoclonal antibody clone 9D5 is more sensitive than the established clone J2 but has not been validated in formalin‐fixed paraffin‐embedded (FFPE) tissue. This study aimed to test and compare the performance of the anti‐dsRNA monoclonal antibodies, 9D5 and J2, in FFPE tissue using an automated staining platform. Archived clinical tissue samples with viral infections (n = 34) and uninfected controls (n = 30) were examined. Immunohistochemical staining for dsRNA (9D5 and J2) and virus‐specific epitopes was performed. 9D5 provided a similar staining pattern but a higher signal‐to‐noise ratio than J2. The following proportions of virus‐infected tissue samples were dsRNA‐positive: SARS‐CoV‐2 (5/5), HPV (6/6), MCV (5/5), CMV (5/6), HSV (4/6), and EBV (0/6). Also, 18 of 30 uninfected samples were dsRNA positive, and an association between fixation time and intensity was observed. However, signals in all samples were markedly reduced by pretreatment with dsRNA‐specific RNAse‐III, indicating a specific reaction. In conclusion, dsRNA can be demonstrated in most viral infections with immunohistochemistry in FFPE tissue but with low clinical specificity. The antibody clone 9D5 performs better than clone J2.
ISSN:0903-4641
1600-0463
DOI:10.1111/apm.13300