Abstract
The Federation of European Laboratory Animal Science Association (FELASA) recommends screening of laboratory rodents and biological materials for a broad variety of bacterial agents, viruses, and parasites. Methods commonly used to date for pathogen detection are neither cost-effective nor time- and animal-efficient or uniform. However, an infection even if silent alters experimental results through changing the animals’ physiology and increases inter-individual variability. As a consequence higher numbers of animals and experiments are needed for valid and significant results. We developed a novel high-throughput multiplex assay, called rodent DNA virus finder (rDVF) for the simultaneous identification of 24 DNA viruses infecting mice and rats. We detected all 24 DNA viruses with high specificity and reproducibility. Detection limits for the different DNA viruses varied between 10 and 1000 copies per PCR. The validation of rDVF was done with DNA isolated from homogenised organs amplified by pathogen specific primers in one multiplex PCR. The biotinylated amplicons were detected via hybridisation to specific oligonucleotide probes coupled to spectrally distinct sets of fluorescent Luminex beads. In conclusion, rDVF may have the potential to replace conventional testing and may simplify and improve routine detection of DNA viruses infecting rodents.
Highlights
Even silent infections of laboratory rodents with a broad variety of DNA viruses can affect research results
Animal Material Since laboratory animals are rarely infected with DNA viruses, mice and 16 rats with a potentially higher infection rate were bought from 12 pet shops in Germany in order to validate the rodent DNA virus finder (rDVF) assay
Detection Limit (DL) Ten-fold endpoint dilution series of plasmids containing the target virus sequence were prepared in the presence of 100 ng/mL murine DNA and analysed by rDVF
Summary
Even silent infections of laboratory rodents with a broad variety of DNA viruses can affect research results. The impact of infections on experimental data may lead to misinterpretation of results and may be responsible for lack of reproducibility. An increased number of animals must be used per experiment to obtain valid and significant data [4]. Standardised laboratory animal health monitoring is a prerequisite and recommended by international organisations such as FELASA [5]. The first FELASA recommendations have been published already 19 years ago [6], there is no uniform program available nor performed in each institute. A comparison of new assays can only be done with established tests used in an institute performing routine health monitoring
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
