The reactivity of native bovine conglutinin (Kg) with antibody against recombinant Kg (rKg), with deletion of the N-terminal and collagen-like regions of the native Kg molecule, was studied by ...sandwich enzyme-linked immunosorbent assay. With anti-recombinant Kg antibody as the coating antibody, rKg reacted with biotinylated homologous anti-rKg and heterologous anti-Kg antibodies as probing antibodies, while native Kg did not. With anti-native Kg antibody as coating antibody, native Kg reacted with biotinylated homologous antibody as probing antibody, while recombinant Kg reacted weakly with both biotinylated homologous and heterologous antibodies. Consequently the N-terminal and collagen-like regions of native Kg molecule are essential to express the complete immunogenicity and/or antigenicity of the native Kg molecule
We isolated porcine cytomegaloviruses (PCMVs) from lung samples of fattening pigs collected in the slaughter houses of 4 prefectures of Japan. Seven isolates were obtained and used for a comparison ...of serological characteristics by ELISA. J1, the first field isolate in Japan, and B6 which was isolated in the UK were also used in the study. The serological relationships between the isolates were analysed by the method of Archetti and Horsfall. OF1 showed serological differences with Chiba2 and Hiroshima. Differences were also observed between Chiba2 and ChibaC, ChibaC and Kagawa. B6 showed differences with OF2, Chiba3, ChibaC and Hiroshima.
A monoclonal antibody (B9) was generated by using a rat malignant fibrous histiocytoma (MFH)-derived cloned cell line (MT-8) as the immunogen. Immunohistochemically, B9 reacted specifically with a ...cytoplasmic antigen of MT-8 cells. Furthermore, B9 immunolabeled another MFH-derived cloned cells (MT-9) and histiocytic sarcoma cells, as well as macrophages/histiocytes in normal and diseased tissues of rats. These findings suggest the presence of a common antigen recognized by B9 between MFH cells and macrophages/histiocytes. This suggests that MFH cells may express histiocytic nature.
Sandwich enzyme-liked immunosorbent assay (ELISA) was developed for the quantitative estimation of Clostridium perfringens enterotoxin (CPE) with monoclonal and polyclonal antibodies as capturing and ...detecting antibodies, respectively. The dose-dependent relationship between absorbance 405 nm and concentration of purified CPE was obtained over the range of 0.64-400 ng/ml. The sandwich ELISA was found to detect crude CPE in culture and CPE in 10% fecal extracts. This method is convenient, rapi
To develop a rapid and specific method to detect and/or identify enterohemorrhagic Escherichia coli O157:H7, two mouse monoclonal antibodies (MAbs) were prepared. Specificities of these two MAbs (1D9 ...and 3E8) were determined by flow cytometry method (FCM). MAbs 3E8 and 1D9 were found to react with E. coli O157:H7, Citrobacter freundii and Salmonella group N (O:30), but not with Escherichia hermannii. With a mixture containing strains of E. coli O157:H7 and E. coli O6:H1, two different peaks appeared in FCM with MAbs, whereas a single peak appeared with polyclonal rabbit antiserum. From these findings, FCM with MAb is suggested to be a rapid, specific, and useful method to detect and identify strains(s) of E. coli O157:H7 in food ingredients
A monoclonal antibody (MAb) reactive with 36 field isolates and 2 laboratory strains of feline calicivirus (FCV) was produced by immunizing mice with the mixture of FCVs. The MAb (4D7) reacted with ...FCVs in an enzyme-linked immunosorbent assay (ELISA), but had no neutralizing activity against the F4 strain of FCV. MAb 8G1, previously produced against the FCV F4 strain, also reacted in ELISA with all FCVs used in the present study. However, the epitopes recognized by 4D7 and 8G1 were different. Using these two MAbs and a polyclonal rabbit antibody, we attempted to develop a sandwich ELISA for detection of FCV antigen. The combination of 4D7 and the polyclonal rabbit IgG was most sensitive. Using this system, all the field isolates of FCV cultured in vitro were detected. However, among the 36 swab samples, from which FCV was isolated, 4 were negative.
Enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against porcine cytomegalovirus (PCMV) was developed using the OF-1 strain of PCMV as an antigen. Results of the ELISA were ...compared to those of indirect fluorescent antibody (IFA) and serum neutralization (SN) tests. ELISA and IFA test were found to be extremely sensitive more than SN test. All of 11 tested sera were highly reactive in both ELISA and IFA test, but 6 of them were antibody-negative in the SN test. The retrospective survery of 436 fattening pig sera collected in Japan in 1981 showed that 433 (99.4%) of them were highly antibody-positive in ELISA.
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