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Animal Bioresource in Japan

.: Home > Animal Bioresource in Japan > 2010 > Volume 59 Number 1 > Yota KITAGAWA1), Yukinobu TOHYA2), Fumio IKE3), Ayako KAJITA3), Sang-Jin PARK1), Yoshiyuki ISHII1), Shigeru KYUWA1) and Yasuhiro YOSHIKAWA1)

Indirect ELISA and Indirect Immunofluorescent Antibody Assay for Detecting the Antibody against Murine Norovirus S7 in Mice

Yota KITAGAWA1), Yukinobu TOHYA2), Fumio IKE3), Ayako KAJITA3), Sang-Jin PARK1), Yoshiyuki ISHII1), Shigeru KYUWA1) and Yasuhiro YOSHIKAWA1)
1) Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo 2) Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo 3) Experimental Animal Division, RIKEN BioResource Center
Abstract :

To evaluate murine norovirus (MNV) infection in laboratory mice, we attempted to develop an enzyme-linked immunosorbent assay (ELISA) system and an indirect immunofluorescent antibody (IFA) assay for detecting the anti-MNV-S7 antibody in mice. MNV-S7, which was isolated in Japan, was used in both assays. The antigen for ELISA was prepared by ultracentrifugation of culture supernatants of RAW 264 cells infected with MNV-S7. Positive sera were obtained from 6-week-old, female C57BL/6JJcl mice inoculated orally with MNV-S7. IFA against infected RAW 264 cells was able to discriminate positive sera from negative sera. Indirect ELISA was performed using 96-well ELISA plates coated with formalin-treated MNV-S7 antigen. In this ELISA system, mouse sera obtained 2 weeks after infection or later showed significantly high OD values and were judged positive. An equal level of anti-MNV-S7 antibody response was observed in BALB/cAJcl, C57BL/6JJcl, DBA/2JJcl, and Jcl:ICR mice; whereas, C3H/HeJJcl mice demonstrated slightly lower antibody production 4 weeks after infection. We also used this ELISA system to evaluate 77 murine serum samples obtained from 15 conventional mouse rooms in research facilities in Japan and found that approximately half of the serum samples contained antibody to MNV-S7. We found that some serum samples were negative for antibodies to mouse hepatitis virus and Mycoplasma pulmonis but positive for antibody to MNV-S7. The results suggest that the MNV infection is more prevalent than other infections such as mouse hepatitis virus and Mycoplasma pulmonis in conventional mouse colonies in Japan, as is the case in other areas of the world.

Keywords :
enzyme-linked immunosorbent assay, immunofluorescent antibody assay, murine norovirus

Date Deposited : 21 Jun 2011 12:59

Last Modified : 21 Jun 2011 14:18

Official URL: http://www.jstage.jst.go.jp/browse/expanim/60/1/_contents

Volume 59, Number 1, - 2010

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