Abstract
Laboratory mice play a tremendous role in biomedical research in studies on immunology, infection, cancer and therapy. In the course of standardization of mice used in animal experiments, health monitoring constitutes an important instrument towards microbiological standardization. Infections with murine astroviruses (MuAstV) were only recently discovered and are, therefore, still relatively unknown in laboratory animal science. In rodent health monitoring viral infections within a population are commonly assessed in terms of specific antibodies by serological testing, as active infection and excretion of virus is often temporary and can easily be missed. So far only ongoing infections with astroviruses can be detected by PCR. The objective of this work was the development of a sensitive and specific MuAstV multiplex serological assay with a high-throughput capability to be used in routine testing of laboratory mice. Four different MuAstV proteins were recombinantly expressed and used as antigens. The best reacting antigen, the capsid spike protein VP27, was selected and tested with a panel of 400 sera of mice from units with a known MuAstV status. Assay sensitivity and specificity resulted in 98.5% and 100%, respectively, compared to RT-PCR results. Eventually this assay was used to test 5529 serum samples in total, during routine diagnostics at the German Cancer Research Center (DKFZ) in Heidelberg between 2015 and 2017. High sero-prevalence rates of up to 98% were detected in units with open cages indicating that the virus is highly infectious and circulates within these populations virtually infecting all animals regardless of the mouse strain. In addition, data collected from 312 mice purchased from commercial breeders and from 661 mice from 58 research institutes in 15 countries worldwide allowed the conclusion that MuAstV is widespread in contemporary laboratory mouse populations.
Highlights
Infections with astroviruses, non-enveloped, single-stranded RNA viruses of the family Astroviridae, are a leading cause of gastroenteritis in humans
Rodent-associated astrovirus was first described by Kjeldsberg and Hem in 1985 [5], but only recently raised awareness in laboratory animal science when detected in fecal samples of wild and laboratory mice by use of metagenomic analysis and PCR [6, 7]
In the course of the maturation process VP90 is cleaved by intracellular caspases at the C-terminus to generate VP70 which in turn is required for the release of immature particles from the host cells [17, 19, 20]
Summary
Infections with astroviruses, non-enveloped, single-stranded RNA viruses of the family Astroviridae, are a leading cause of gastroenteritis in humans. 1975 following an outbreak of diarrhea in children [1, 2] Since their first discovery, astroviruses were isolated from a wide range of mammalian and avian species including mink, chicken, cat, dog, deer, duck, turkey, sheep, cattle and bat [3]. Astroviruses were isolated from a wide range of mammalian and avian species including mink, chicken, cat, dog, deer, duck, turkey, sheep, cattle and bat [3] Based on their host tropism and genetic relatedness astroviruses are grouped in two genera, Mamastrovirus and Avastrovirus, infecting mammals and birds, respectively [4]. As a result of the lack of serological detection methods so far only active infections with virus excretion can be detected by PCR and prevalence and distribution within laboratory mice populations are unclear
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