Development of ELISA using recombinant antigens for specific detection of mouse parvovirus infection.
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.
Kunita
Satoshi
S
Laboratory Animal Resource Center, University of Tsukuba, Ibaraki, Japan.
Chaya
Miyuki
M
Hagiwara
Kozue
K
Ishida
Tomoko
T
Takakura
Akira
A
Sugimoto
Tatsuya
T
Iseki
Hiroyoshi
H
Fuke
Kumiko
K
Sugiyama
Fumihiro
F
Yagami
Ken-ichi
K
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D036022:Capsid Proteins / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051379:Mice; D008915:Minute Virus of Mice / Q000276:immunology; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D017992:Parvovirus / Q000276:immunology*; D011994:Recombinant Proteins / Q000032:analysis; D012376:Rodent Diseases / Q000175:diagnosis* / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology
D000956:Antigens, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins