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Updated: Aug 15, 2026
Plaque Assay for Murine Norovirus
Published on: August 22, 2012
Development of a microsphere-based serologic multiplexed fluorescent immunoassay and a reverse transcriptase PCR
Charlie C Hsu1, Christiane E Wobus, Earl K Steffen
1Research Animal Diagnostic Laboratory, Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, E108 Veterinary Medicine Building, 1600 E. Rollins Road, Columbia, MO 65211, USA. hsuc@missouri.edu
Abstract:
Murine norovirus 1 (MNV-1) is a newly recognized pathogen of mice that causes lethal infection in mice deficient in components of the innate immune response but not in wild-type 129 mice. In this study, in vitro-propagated MNV-1 was used as antigen to develop a multiplexed fluorescent immunoassay (MFI) to detect antibodies to MNV-1 in infected mice. The MNV-1 MFI was 100% specific and 100% sensitive in detecting anti-MNV-1 antibody in sera from experimentally infected mice. Testing of a large number of mouse serum samples (n = 12,639) submitted from contemporary laboratory mouse colonies in the United States and Canada revealed that 22.1% of these sera contained antibodies to MNV-1, indicating infection with MNV-1 is widespread in research mice. In addition, a reverse transcriptase PCR primer pair with a sensitivity of 25 virus copies was developed and used to demonstrate that MNV-1 RNA could be detected in the spleen, mesenteric lymph node, and jejunum from some experimentally infected mice 5 weeks postinoculation. These diagnostic assays provide the necessary tools to define the MNV-1 infection status of research mice and to aid in the establishment of laboratory mouse colonies free of MNV-1 infection.
Insights
Murine norovirus 1 (MNV-1) is a common mouse pathogen. New diagnostic tools, including a multiplexed fluorescent immunoassay (MFI) and RT-PCR, can detect MNV-1 infection in research mice.
Area of Science:
- Veterinary Virology
- Immunology
- Infectious Diseases
Background:
- Murine norovirus 1 (MNV-1) is an emerging mouse pathogen.
- MNV-1 causes lethal disease in immunocompromised mice but is often asymptomatic in wild-type mice.
- The prevalence and diagnostic methods for MNV-1 in research settings require further investigation.
Purpose of the Study:
- To develop and validate diagnostic assays for Murine norovirus 1 (MNV-1).
- To assess the prevalence of MNV-1 infection in laboratory mouse colonies.
- To provide tools for establishing MNV-1-free mouse colonies.
Main Methods:
- Development of a multiplexed fluorescent immunoassay (MFI) using in vitro-propagated MNV-1 as antigen.
- Validation of MFI specificity and sensitivity using experimentally infected mouse sera.
- Development of a sensitive reverse transcriptase PCR (RT-PCR) assay for MNV-1 RNA detection.
- Screening of 12,639 mouse serum samples from US and Canadian colonies using MFI.
- Detection of MNV-1 RNA in tissues of experimentally infected mice via RT-PCR.
Main Results:
- The MNV-1 MFI demonstrated 100% specificity and 100% sensitivity for detecting anti-MNV-1 antibodies.
- Antibodies to MNV-1 were detected in 22.1% of 12,639 tested mouse sera, indicating widespread infection.
- MNV-1 RNA was detectable in spleen, mesenteric lymph node, and jejunum up to 5 weeks postinoculation in experimentally infected mice.
Conclusions:
- MNV-1 infection is prevalent in research mouse populations.
- Developed MFI and RT-PCR assays are effective tools for diagnosing MNV-1 infection.
- These assays will aid in establishing and maintaining MNV-1-free research mouse colonies.
