AbstractMouse hepatitis virus (MHV) is a common infection in a large number of laboratory mouse colonies and is known to interfere with research results. Serology testing is a good method of detecting the historical presence of MHV, however, it cannot detect when the mouse is currently shedding the virus.
The purpose of this study was to develop an in-house Polymerase Chain Reaction (PCR) assay to detect MHV in naturally infected mice using faecal samples. PCR is a highly sensitive test that can detect few copies of virus in a given sample. The PCR was also used to detect the length of time MHV was shedding in C57BL6/J mice.
A number of problems with cross-contamination were encountered and largely overcome. Standard conditions for extracting RNA from faecal pellets, reverse-transcribing it into cDNA and detecting the presence of viral sequences by PCR were developed.
The pattern of viral shedding from “naïve” mice introduced into the animal facility was found to be variable, even between animals housed in the same cage. Not all animals appeared to shed virus at all and, of those that did, some showed more than one cycle of shedding within the observation period.
Shedding was more consistent from post-weaning animals born from matings between members of the introduced cohort, but still with some variability. Not all animals had apparently cleared the virus at the end of the 30-day observation period.
There was generally good correlation between the detection of viral sequences and the ultimate serological status of sample mice, with some notable exceptions.
These results are discussed in the context of using PCR to evaluate the current “viral “load” in an animal facility and in developing strategies for elimination of the organism.
|Date of Award||2015|
|Supervisor||Colin Henderson (Supervisor) & Edward Newman (Supervisor)|