Antemortem detection of mouse parvovirus and mice minute virus by polymerase chain reaction (PCR) of faecal samples
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Published on: July 6, 2013
1Research Animal Diagnostic Laboratory, University of Missouri, Columbia, MO 65211, USA. bauerbe@missouri.edu
Abstract:
Parvoviruses remain one of the most common viral infections seen in laboratory mouse colonies. The purpose of this study was to develop an antemortem polymerase chain reaction (PCR) assay to detect mice infected with mouse parvovirus-1 (MPV) and mice minute virus (MMV) using faecal samples. The MMV PCR assay consistently detected as few as 100 plasmid copies of MMV in faecal samples, while the MPV PCR assay detected as few as 10 plasmid copies of MPV. Faecal pellets from infected mice held at room temperature from 1 to 7 days tested positive by MMV and MPV PCR, respectively. This demonstrates that parvovirus DNA is stable in faecal samples kept at room temperature. PCR assays were also used to follow the length of MMV and MPV shedding in faeces from SENCAR mice, which were endemically infected with multiple agents. MMV faecal shedding was detected in 60-70% of the mice 5-7 weeks old, and by 13 weeks of age, faecal samples from all mice were negative for MMV. MPV faecal shedding was detected in 90-100% of the mice 5-11 weeks old; however, by 19 weeks of age, faecal samples from all mice were negative for MPV. These findings confirm that faecal shedding occurs for a limited time and suggest that 5-9-week-old mice are the most appropriate age group in endemically infected mice for faecal testing by MMV and MPV PCR.
Insights
Develop a sensitive polymerase chain reaction (PCR) assay to detect mouse parvovirus-1 (MPV) and minute virus (MMV) in mouse feces. This assay is stable at room temperature and aids in identifying shedding periods for optimal testing ages.
Area of Science:
- Veterinary Virology
- Laboratory Animal Science
- Molecular Diagnostics
Background:
- Parvoviruses, including mouse parvovirus-1 (MPV) and minute virus of mice (MMV), are prevalent in laboratory mouse colonies.
- Accurate and timely detection of these viruses is crucial for maintaining colony health and research integrity.
Purpose of the Study:
- To develop and validate an antemortem polymerase chain reaction (PCR) assay for detecting MPV and MMV in fecal samples.
- To assess the stability of parvovirus DNA in fecal samples stored at room temperature.
- To determine the duration of fecal shedding for MPV and MMV in endemically infected mice.
Main Methods:
- Development of specific PCR assays for MPV and MMV detection.
- Quantification of viral DNA detection limits using plasmid standards.
- Evaluation of DNA stability in fecal samples stored at room temperature for up to 7 days.
- Longitudinal monitoring of MPV and MMV fecal shedding in naturally infected SENCAR mice.
Main Results:
- The MPV PCR assay detected as few as 10 plasmid copies, and the MMV PCR assay detected as few as 100 plasmid copies.
- Parvovirus DNA remained stable in fecal samples stored at room temperature for 1 to 7 days.
- MMV shedding was observed in 60-70% of mice aged 5-7 weeks, clearing by 13 weeks.
- MPV shedding was detected in 90-100% of mice aged 5-11 weeks, clearing by 19 weeks.
Conclusions:
- A sensitive and stable fecal PCR assay for MPV and MMV has been developed.
- Fecal samples are suitable for parvovirus detection at room temperature for up to 7 days.
- Optimal fecal testing ages for MPV and MMV in endemically infected mice are 5-9 weeks due to limited shedding duration.
