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Development of ELISA using recombinant antigens for specific detection of mouse parvovirus infection
Satoshi Kunita1, Miyuki Chaya, Kozue Hagiwara
1Laboratory Animal Resource Center, University of Tsukuba, Ibaraki, Japan.
Abstract:
Nucleotide sequences of mouse parvovirus (MPV) isolate, named MPV/UT, and mouse minute virus (MMV) were analyzed and used for expressing recombinant proteins in E. coli. ELISA tests using recombinant major capsid protein (rVP2) and recombinant major non-structural protein (rNS1) as antigens were developed and their performance in serologic detection of rodent parvovirus infection was assessed. MPV-rVP2 and MMV-rVP2 ELISAs reacted specifically with anti-MPV and anti-MMV mouse sera, respectively. MMV-rNS1 antigen had a wide reaction range with antisera to rodent parvoviruses including MPV, MMV, Kilham rat virus (KRV) and H-1 virus. All mice oronasally infected with MPV were seropositive at 4 weeks post-infection in screening by ELISAs using MPV-rVP2 and MMV-rNS1 antigens, but were negative by conventional ELISA using whole MMV antigen. A contact transmission experiment revealed that transmission of MPV occurred up to 4 weeks post-infection, and all cage mates were seropositive in screening with MPV-rVP2 and MMV-rNS1 ELISAs. These results indicate that MPV-rVP2 and MMV-rVP2 are specific ELISA antigens which distinguish between MPV and MVM infection, while MMV-rNS1 antigen can be used in generic ELISA for a variety of rodent parvoviruses. The higher sensitivity of MPV-rVP2 ELISA than conventional ELISA for detecting seroconversion to MPV in oronasally infected mice as well as in cage mates suggests the usefulness of MPV-rVP2 ELISA in quarantine and microbiological monitoring of MPV infection in laboratory mice.
Insights
New ELISAs using recombinant proteins detect mouse parvovirus (MPV) and mouse minute virus (MMV) infections. MPV-rVP2 ELISA specifically detects MPV, while MMV-rNS1 detects multiple rodent parvoviruses, improving laboratory mouse monitoring.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Rodent parvoviruses, including mouse parvovirus (MPV) and mouse minute virus (MMV), pose challenges in laboratory animal health.
- Accurate serological detection methods are crucial for monitoring and controlling parvovirus infections in research settings.
Purpose of the Study:
- To develop and evaluate novel Enzyme-Linked Immunosorbent Assays (ELISAs) using recombinant viral proteins for the specific and broad detection of rodent parvoviruses.
- To assess the sensitivity and specificity of these new ELISAs compared to conventional methods for diagnosing MPV infection.
Main Methods:
- Nucleotide sequences of MPV and MMV were analyzed for recombinant protein expression in E. coli.
- Recombinant major capsid protein (rVP2) and non-structural protein (rNS1) were produced and used as antigens in ELISA development.
- ELISA performance was assessed using specific and cross-reactive rodent parvovirus antisera and experimentally infected mice.
Main Results:
- MPV-rVP2 and MMV-rVP2 ELISAs demonstrated specificity for MPV and MMV infections, respectively.
- MMV-rNS1 ELISA showed broad reactivity against various rodent parvoviruses, including MPV, MMV, Kilham rat virus (KRV), and H-1 virus.
- ELISAs utilizing MPV-rVP2 and MMV-rNS1 antigens detected MPV infection in experimentally infected mice and their cage mates more effectively than conventional ELISA.
Conclusions:
- Recombinant MPV-rVP2 and MMV-rVP2 proteins serve as specific antigens for differentiating MPV and MMV infections.
- Recombinant MMV-rNS1 is a valuable antigen for a generic ELISA detecting a range of rodent parvoviruses.
- The high sensitivity of MPV-rVP2 ELISA makes it a powerful tool for MPV detection in quarantine and microbiological monitoring of laboratory mice.
