title: Reverse transcription-polymerase chain reaction detection and nucleic acid sequence confirmation of reovirus infection in laboratory mice with discordant serologic indirect immunofluorescence assay and enzyme-linked immunosorbent assay results.
abstract: Reovirus infections are typically subclinical in weaned mice, and are best detected using serologic tests. After exposure to the soiled bedding of some mice obtained from various sources, numerous sentinel mice tested reovirus seropositive by use of indirect immunofluorescence assays (IIFA) in our institution. A major commercial rodent pathogen testing laboratory verified our IIFA results, but since the same samples were reovirus seronegative using their "more specific" enzyme-linked immunosorbent assay (ELISA), the IIFA results were reported as "false positives." As past in-house observations suggested transmission of the virus to sentinel and other animals, we sought to determine whether the IIFA results were always "false positives." An opportunity to test this notion arose after receipt of reovirus IIFA-positive transgenic mice from an academic source. Using reverse transcriptase-polymerase chain reaction (RT-PCR) assays, the presence of reovirus RNA was detected in fecal specimens taken from some sentinel animals that subsequently seroconverted from IIFA-negative to IIFA-positive for reovirus. The virus could not be isolated by use of tissue culturing methods. Nucleotide sequence analysis established the presence of unique reovirus sequences. These results indicate that contemporary reovirus infections may not be detected by use of some serologic tests, and that RT-PCR analysis may be useful for confirmation of active reovirus infection in certain situations.
authors.affiliation: Department of Comparative Medicine, Loyola University Medical Center, Maywood, Illinois 60153, USA.
authors.forename: Mina Hicks
authors.initials: MH
authors.lastname: Wright
authors.forename: Lee M
authors.initials: LM
authors.lastname: Cera
authors.forename: Nicolene A
authors.initials: NA
authors.lastname: Sarich
authors.forename: John A
authors.initials: JA
authors.lastname: Lednicky
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005189:False Positive Reactions; D005243:Feces / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D006799:Housing, Animal; D012089:Mammalian orthoreovirus 3 / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D012088:Reoviridae Infections / Q000097:blood / Q000635:transmission*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018571:Sentinel Surveillance; D016415:Sequence Alignment