title: Development of a microsphere-based serologic multiplexed fluorescent immunoassay and a reverse transcriptase PCR assay to detect murine norovirus 1 infection in mice.
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.
authors.affiliation: Research 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
authors.forename: Charlie C
authors.initials: CC
authors.lastname: Hsu
authors.forename: Christiane E
authors.initials: CE
authors.lastname: Wobus
authors.forename: Earl K
authors.initials: EK
authors.lastname: Steffen
authors.forename: Lela K
authors.initials: LK
authors.lastname: Riley
authors.forename: Robert S
authors.initials: RS
authors.lastname: Livingston
mesh_terms: D000818:Animals; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000097:blood; D017250:Caliciviridae Infections / Q000175:diagnosis*; D002170:Canada; D015200:Fluoroimmunoassay / Q000379:methods*; D051379:Mice / Q000821:virology*; D008863:Microspheres; D029322:Norovirus / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014018:Tissue Distribution; D014481:United States
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D012367:RNA, Viral