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Updated: Mar 24, 2026
Detection of Viral RNA by Fluorescence in situ Hybridization FISH
Published on: May 5, 2012
Specific Detection of Two Divergent Simian Arteriviruses Using RNAscope In Situ Hybridization
Shu Qìng Yú1, Yíngyún Caì1, Cassandra Lyons1
1Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick, Frederick, Maryland, United States of America.
Abstract:
Simian hemorrhagic fever (SHF) is an often lethal disease of Asian macaques. Simian hemorrhagic fever virus (SHFV) is one of at least three distinct simian arteriviruses that can cause SHF, but pathogenesis studies using modern methods have been scarce. Even seemingly straightforward studies, such as examining viral tissue and cell tropism in vivo, have been difficult to conduct due to the absence of standardized SHFV-specific reagents. Here we report the establishment of an in situ hybridization assay for the detection of SHFV and distantly related Kibale red colobus virus 1 (KRCV-1) RNA in cell culture. In addition, we detected SHFV RNA in formalin-fixed, paraffin-embedded tissues from an infected rhesus monkey (Macaca mulatta). The assay is easily performed and can clearly distinguish between SHFV and KRCV-1. Thus, if further developed, this assay may be useful during future studies evaluating the mechanisms by which a simian arterivirus with a restricted cell tropism can cause a lethal nonhuman primate disease similar in clinical presentation to human viral hemorrhagic fevers.
Insights
A new in situ hybridization assay detects Simian hemorrhagic fever virus (SHFV) RNA in cell cultures and infected rhesus monkey tissues. This method aids in studying lethal nonhuman primate diseases caused by simian arteriviruses.
Area of Science:
- Veterinary Virology
- Nonhuman Primate Pathology
- Molecular Diagnostics
Background:
- Simian hemorrhagic fever (SHF) is a lethal disease in Asian macaques, often caused by Simian hemorrhagic fever virus (SHFV).
- Pathogenesis studies for SHFV are limited by a lack of standardized reagents for detecting viral tropism.
- Understanding SHFV pathogenesis is crucial for comparative studies with human viral hemorrhagic fevers.
Purpose of the Study:
- To establish a reliable in situ hybridization (ISH) assay for detecting SHFV RNA.
- To validate the assay's ability to differentiate SHFV from other simian arteriviruses, such as Kibale red colobus virus 1 (KRCV-1).
- To demonstrate the assay's utility in detecting SHFV in infected nonhuman primate tissues.
Main Methods:
- Development of an in situ hybridization assay for SHFV and KRCV-1 RNA detection.
- Application of the assay to detect viral RNA in cultured cells.
- Testing the assay on formalin-fixed, paraffin-embedded tissues from a SHFV-infected rhesus macaque.
Main Results:
- The established ISH assay successfully detected SHFV RNA in cell culture.
- The assay accurately distinguished between SHFV and KRCV-1 RNA.
- SHFV RNA was successfully detected in formalin-fixed, paraffin-embedded rhesus macaque tissues.
Conclusions:
- The developed ISH assay is a valuable tool for detecting SHFV and KRCV-1 RNA.
- This assay facilitates future research into the pathogenesis of SHFV and related simian arteriviruses.
- The assay's ability to detect viral RNA in fixed tissues supports in vivo tropism studies for lethal nonhuman primate diseases.
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