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. 2021 Dec 17;11:24191. doi: 10.1038/s41598-021-03638-5

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Comparison of distribution of Usutu virus (USUV) specific antigen and double-stranded ribonucleic acid (dsRNA) expression in the spleen between two animals with qRT-PCR confirmed USUV infection with (A-D) and without (E–H) immunohistochemical USUV antigen detection. (A) Moderate amounts of USUV antigen-labeled cells in the spleen of an animal (Table 2, #1) with USUV confirmed infection by qRT-PCR. (B) In contrast, only few cells labeled positive for dsRNA by applying the 9D5 antibody (arrowheads). Inset: 9D5-labeled cells at higher magnification. A moderate number of cells showed positive reaction with dsRNA-specific antibodies K1 (C, arrowheads) and J2 (D, arrowheads) similar to the USUV antigen distribution with respect to both, amount and distribution of immunopositive cells. Inset: K1- and J2-labeled cells at higher magnification. (E) Despite lack of viral antigen the spleen of a blackbird with qRT-PCR-confirmed USUV infection (Table 2, #7) was tested positive for dsRNA by the application of antibodies directed against dsRNA. Although, the amount of immunopositive cells varied substantially: Detection level of dsRNA by using the 9D5 antibody (F, arrowheads) remained low in comparison to K1 (G, arrowhead) and J2 (H, arrowhead) antibody, respectively. Insets: 9D5-, K1- and J2-labeled cells at higher magnification. (scale bar: 50 µm; immunohistochemistry).