Development of an in situ hybridization assay using an AS1 probe for detection of bovine leukemia virus in BLV-induced l
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ORIGINAL ARTICLE
Development of an in situ hybridization assay using an AS1 probe for detection of bovine leukemia virus in BLV‑induced lymphoma tissues Kiyohiko Andoh1 · Kumiko Kimura2 · Asami Nishimori1 · Shinichi Hatama1 Received: 3 August 2020 / Accepted: 7 September 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Enzootic bovine leukosis (EBL) is a malignant B cell lymphoma caused by infection with bovine leukemia virus (BLV). Histopathological examination is commonly used for diagnosis of the disease, but observation of lymphoma alone does not confirm EBL because cattle may be affected by sporadic forms of lymphoma that are not associated with BLV. Detection of BLV in tumor cells can be definitive evidence of EBL, but currently, there is no technique available for such a purpose. In this study, we focused on a viral non-coding RNA, AS1, and developed a novel in situ hybridization assay for the detection of BLV from formalin-fixed paraffin-embedded (FFPE) tissues. RNA-seq analysis revealed that all examined B lymphocytes derived from clinical EBL abundantly expressed AS1 RNA, indicating a possible target for detection. The in situ hybridization assay using an AS1 probe clearly detected AS1 RNA in fetal lamb kidney cells persistently infected with BLV. The utility of this assay in clinical samples was assessed using three EBL-derived lymph node specimens and one BLV-negative specimen, and AS1 RNA was detected specifically in the EBL-derived tissues. These results suggest that AS1 RNA is a useful target for the detection of BLV from FFPE specimens of tumor tissues. This technique is expected to become a powerful tool for EBL diagnosis.
Introduction Handling Editor: William G Dundon. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00705-020-04837-7) contains supplementary material, which is available to authorized users. * Kumiko Kimura [email protected] * Shinichi Hatama [email protected] Kiyohiko Andoh [email protected] Asami Nishimori [email protected] 1
Division of Viral Disease and Epidemiology, National Institute of Animal Health, National Agriculture and Food Research Organization, 3‑1‑5 Kannondai, Tsukuba, Ibaraki 305‑0856, Japan
Division of Pathology and Pathophysiology, National Institute of Animal Health, National Agriculture and Food Research Organization, 3‑1‑5 Kannondai, Tsukuba, Ibaraki 305‑0856, Japan
2
Bovine lymphoma is a fatal neoplastic disease of lymphoid cells in cattle and is divided into two types, enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL). EBL is a malignant B cell lymphoma caused by infection with bovine leukemia virus (BLV), a member of family Retroviridae, genus Deltaretrovirus. Although the majority of BLV-infected cattle remain healthy and show no clinical symptoms, one-third of infected cattle develop persistent lymphocytosis characterized by non-malignant polyclonal B cell expansion, and a small fraction (1-5%) of infected cattle develop EBL [1, 3, 6, 16]
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