Reovirus 3 PCR test for rodents

Reovirus type 3 (Reo-3) can infect many different rodent species and is prevalent in laboratory rodent colonies. It is a common contaminant of transplantable tumor cell lines and other biological materials.

Infection of neonatal mice with Reo-3 can lead to a multi-systematic disease that is characterized by necrotizing hepatitis, myocarditis, pancreatitis, and meningoencephalitis. Steatorrhoea often develops secondary to liver disease and results in “oily skin” appearance.

Reo-3 virus can be transmitted between animals by aerosol and fecal–oral routes, and fomite transmission is possible because the virus is environmentally stable. One infected animal can easily lead to rapid outbreak of the disease and therefore rapid diagnosis is essential to prevent transmission through an animal facility.

Several methods are currently employed to detect Reo-3 infection in rodents and contaminated biological materials. Serological detection of anti-viral antibodies has typically been used to diagnose Reo-3 infection in rodents. However, serological assays cannot detect virus infections directly in immunodeficient strains of rodents that do not generate a humoral immune response, and even in immunocompetent rodents the time needed for seroconversion is too long for rapid diagnosis of the disease.

Molecular detection by real time PCR, on the other hand, has proven to be a useful technique for detection of Reo-3 (Steele, 1995), as it is highly sensitive and also very specific. Unlike serology testing, there is no cross-reactivity with other similar viruses. Real time PCR is also an attractive alternative to the rodent antibody production (RAP) test for detection of Reo-3 contamination in biological materials (Compton & Riley 2001). Compared to RAP testing, real time PCR offers faster turnaround time and lower cost.

Utilities:

Confirm the disease causing agent
Shorten the time required to confirm a clinical diagnosis of Reo-3
Ensure that vivariums are free of Reo-3
Early prevention of spread of this virus among a vivarium
Minimize personnel exposure to this virus
Safety monitoring of biological products that derive from mice

References:
Compton, S.R. and Riley, L.K. (2001) Detection of infectious agents in laboratory rodents: traditional and molecular techniques. Comparative Medicine 51:113–119.
Steele, M.I. (1995) Reovirus 3 not detected by reverse transcriptase-mediated polymerase chain reaction analysis of preserved tissue from infants with cholestatic liver disease. Hepatology 21: 697–702.

Specimen requirements: Nasopharyngeal or tracheal swab, or fecal pellet, or fresh or paraffin-embedded tissue, shipped overnight at room temperature; or tissue shipped frozen.

For specimen types other than those listed here, please call to confirm specimen acceptability and shipping instructions.

For all specimen types, if there will be a delay in shipping, or during very warm weather, refrigerate specimens until shipped and ship with a cold pack unless more stringent shipping requirements are specified. Frozen specimens should be shipped so as to remain frozen in transit. See shipping instructions for more information.

Turnaround time: 2 business days

Methodology: Ultrasensitive qualitative reverse transcription real time polymerase chain reaction

Normal range: Nondetected