Isolation of reovirus type 3 from foals.
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Biomedical subjects
Publications and source records attributed to M Matumoto.
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Japanese sera were shown by enzyme-linked immunosorbent assay (ELISA) to contain frequently antibodies against allantoic fluid, confirming the results obtained in our previous study on ELISA for the detection of mumps antibodies. Chromatographic analysis with Sephacryl S-300 of the allantoic fluid antigen used for ELISA suggested the antigens for these antibodies in Japanese sera to be macromolecular proteins. Comparative absorption tests with allantoic fluid and ovomacroglobulin were therefore carried out on serum samples positive in ELISA with allantoic fluid antigen, and ovomacroglobulin was identified to be the antigenic component for antibodies to allantoic fluid antigen in Japanese sera.
Transplacental transmission was demonstrated in pregnant mice subcutaneously inoculated with Getah virus. Viremia was shown in the infected dams, and high-titered virus was detected in the placenta and later in the fetus, suggesting virus invasion of the fetus through hematogenous infection of the placenta. High-titered virus was shown in the fetal brain and muscle and in the brain of the young dying soon after birth. Intrauterine infection resulted in a reduction of the litter size, number of young born alive and survival rate to 1 week of age. These results were further corroborated by necropsy performed several days after virus inoculation. The stage of gestation at the time of virus inoculation greatly influenced these results. Dams inoculated at 12 days of gestation delivered all dead babies, whereas virus inoculation at 5 days of gestation had no effect on the number of young born alive. The dams inoculated at 8 days of gestation had reduced litter sizes and those inoculated at 16 days of gestation produced slightly fewer live babies. Gestational stage at the time of virus inoculation also influenced viral growth in fetuses and placentas. The infection rate was low in dams inoculated at 5 days of gestation, high in dams inoculated at 8 or 16 days of gestation and 100% in dams inoculated at 12 days of gestation. High-titered virus was shown in placentas and fetuses of the dams inoculated at 8, 12 or 16 days of gestation. These results suggest that Getah virus may readily cross the placental barrier through hematogenous infection of the placenta in mice.
An enzyme-linked immunosorbent assay (ELISA) was developed for detection of antibody against porcine parvovirus in swine sera. The antigen used for the assay was partially-purified virus treated with fluorocarbon and shown to contain 7 proteins by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Of these proteins 83-, 64- and 60-K proteins reacted in Western immunoblotting with swine serum after infection with porcine parvovirus. Antibody responses were demonstrated by ELISA in pigs subcutaneously-infected with porcine parvovirus as by hemagglutination-inhibition (HI) test and Western immunoblotting reaction with the 83-, 64- and 60-K viral proteins. The results of ELISA on random swine-serum samples were well-correlated with those of the HI test. These findings indicate the usefulness of the ELISA as a serological tool for porcine parvovirus infection.
A new method for the detection of pseudorabies antibody in swine sera was developed by use of the indirect immunoperoxidase plaque staining test. The indirect immunoperoxidase plaque staining test demonstrated serologic responses in pigs that were subcutaneously infected with pseudorabies virus. The test had a sensitivity and a specificity comparable to those of the complement-dependent neutralization test and appeared to be somewhat more sensitive than the neutralization test and the enzyme-linked immunosorbent assay. The test was simple to perform, and the results could be read with the unaided eye under ordinary light, making the test particularly suitable for the routine testing of large numbers of serum samples.
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Mouse myeloma cells (SP2/O) were fused with spleen cells from BALB/c mice immunized with detergent-solubilized antigen of purified virus, and 21 monoclonal (MC) antibodies reactive in enzyme-linked immunosorbent assay with the TO-163 strain of porcine transmissible gastroenteritis (TGE) virus were obtained. Of these MC antibodies, 14, 6 and 1 were IgG1, IgG2a and IgM, respectively. All of the MC antibodies contained light chains of the kappa type. Of these MC antibodies, 8 were found to have neutralization (NT) activity against the TO-163 strain. Comparison of 7 strains of TGE virus by NT tests using our panel of MC antibodies confirmed their close antigenic relationships, but also revealed the occurrence of distinct antigenic differences. These results suggest that there may be at least 6 different epitopes involved in NT reaction on the virion of the TO-163 strain. This notion was confirmed by the competitive binding assay.
Slow-reacting complement-requiring neutralizing (NT) antibody was detected in sera from cattle 2 weeks after infection with Akabane virus. Bovine sera obtained 3 or 4 weeks after infection contained slow-reacting noncomplement-requiring NT antibody. The slow-reacting complement-requiring NT antibody was sensitive to 2-mercaptoethanol (2-ME), whereas the slow-reacting noncomplement-requiring NT antibody was resistant to 2-ME. The initial phase may represent the IgM response and the later phase a change to IgG. A NT test was developed in which virus-serum mixtures were incubated at 4 degrees C for 48 h and then with complement at 37 degrees C for 60 min; this gave an improved sensitivity over the previous incubation at 37 degrees C for 60 min.
The Lincoln strain of bovine rotavirus was found to replicate with cytopathic effects in cultures of GBK cells, a stable cell line derived from bovine kidney, when the cultures were maintained in the presence of trypsin. The virus was readily passaged and the infected cells were shown to contain specific viral antigen by indirect immunofluorescent staining. The virus formed plaques in GBK cell monolayers, when trypsin was incorporated in the agar overlay medium. The plaque count increased about twofold when diethylaminoethyl dextran was further included in the overlay medium. Plaque assay in GBK cells was more sensitive than that in MA-104 cells previously reported by Matsuno et al. The specificity of plaques was confirmed by specific inhibition with antiserum against the Lincoln strain.
Akabane virus was isolated from the biting midge, Culicoides oxystoma, collected in a cowshed in Kagoshima on Kyushu Island of Japan. This is the first report on the isolation of Akabane virus from biting midges of the genus Culicoides in Japan. Two calves kept as bait in the cowshed seroconverted to Akabane virus. These results strongly suggest that C. oxystoma may be a vector of Akabane virus.
An enzyme-linked immunosorbent assay (ELISA) using a detergent-solubilized antigen of purified virus was developed for detection of antibody against porcine transmissible gastroenteritis (TGE) virus in swine serum. The ELISA demonstrated antibody responses in pigs immunized intramuscularly with the attenuated TO-163 strain of TGE virus and in pigs orally infected with the virulent Shizuoka strain of the virus. The results of the ELISA were well correlated with those of the neutralization test. These results indicate the usefulness of the ELISA as a serological tool for TGE virus antibody.
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An outbreak of neonatal calf diarrhea was studied on a breeding farm of Japanese indigenous beef cattle. During the breeding season of 1982, 43 calves were born over the period 27 February-28 April. All but one of the calves suffered from neonatal diarrhea and 5 died. Bovine rotavirus was isolated in cell cultures from fecal specimens of 39 (90.7%) of the 43 calves during the outbreak, strongly suggesting that this was the causative agent; the virus was readily isolated from 81 (83.5%) of 97 specimens of diarrhea. Rotavirus was subsequently isolated from the feces of 7 of the calves in early May, more than one month after the initial virus isolation in early March. Two of these calves were again rotavirus-positive in early June, 41 days after the second virus isolation. Diarrhea had ceased in all 7 calves in March. Some antigenic differences were shown by the neutralization test between the early and later isolates from one of these calves, suggesting either re-infection with a serologically different virus, or persistent infection with the original virus following antigenic modulation.
High-performance liquid chromatography was successfully used to separate hemagglutination-inhibiting immunoglobulin M (IgM) rubella virus antibody from IgG rubella virus antibody in human serum. The fractionation by high-performance liquid chromatography was as effective as sucrose density gradient centrifugation in separating IgM antibody from IgG antibody.