Multiplex polymerase chain reaction assay for the detection of minute virus of mice and mouse parvovirus infections in laboratory mice

Abstract

Mouse parvoviruses are among the most prevalent infectious pathogens in contemporary mouse colonies. To improve the efficiency of routine screening for mouse parvovirus infections, a multiplex polymerase chain reaction (PCR) assay targeting the VP gene was developed. The assay detected minute virus of mice (MVM), mouse parvovirus (MPV) and a mouse housekeeping gene (a-actin) and was able to specifically detect MVM and MPV at levels as low as 50 copies. Co-infection with the two viruses with up to 200-fold differences in viral concentrations can easily be detected. The multiplex PCR assay developed here could be a useful tool for monitoring mouse health and the viral contamination of biological materials.

Non-Invasive Fecal Based PCR Assays for Detection of Mouse Parvo Virus and Minute Virus of Mice in Laboratory Mice

“…as 0.4 fg of the plasmid DNA template in the presence of negative mouse fecal DNA (Fig 1). Bauer & Riley (2006) and Wang et al (2013) stated that the PCR assays detected as low as 100 copies of MVM and 10 copies of MPV and 10 copies of MVM and 50 copies of MPV respectively. In the current study, Plasmid DNA was used for the detection of the sensitivity of each set of primers; hence sensitivity assay was limited to the concentration of Plasmid DNA to 0.4fg, to avoid over-estimating the efficiency of primers.…”

“…Direct screening with PCR detects viruses even in early infections, in immunocompromised mice (Besselsen et al 1995) and cell lines (Wang et al 2013) but employing PCR as a post-mortem testing doesn't justify it as an alternative to serology in health monitoring.…”

“…Fecal based PCR assays offer the advantages of easy sample collection and antemortem testing of valuable animals (Bauer and Riley, 2006). On considering 3R's principle, non-invasive fecal based diagnosis of viral excretion of MPV and MVM is a very promising tool after considering the time frame of sample collection (Janus et al 2008;Macy et al 2009;Wang et al 2013).…”