Enzyme-linked-immunosorbent assay for the detection of antibodies to Fusobacterium necrophorum in cattle with lesions
| Project/Area Number |
02454103
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
基礎獣医学
|
|---|
| Research Institution | Faculty of Agriculture, Yamaguchi University |
|---|
Principal Investigator |
KANOE Masamitsu Department of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (00035099)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
INOUE Makoto Department of Veterinary Pathology, Faculty of Agriculture, Yamaguchi University, 農学部, 助手 (80035112)
KAI Kazushige Department of Veterinary Microbiology, Faculty of Agriculture, Yamaguchi Univers, 農学部, 助教授 (60085628)
|
|---|
| Project Period (FY) |
1990 – 1992
|
| Project Status |
Completed (Fiscal Year 1992)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1992: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥2,700,000 (Direct Cost: ¥2,700,000)
|
|---|
| Keywords | Necrobacillosis / Anaerobic bacteria / Serodiagnosis / ELISA / Hepatic abscess / Predictive diagnosis / Opportunistic infection / 内臓膿瘍 |
|---|
| Research Abstract |
An enzyme-linked-immunosorbent assay (ELISA) with HCl heat extracted antigen of Fusobacterium necrophorum was developed for the detection of antibody in bovine sera. Optimal conditions for antigen concentration and dilution of bovine serum were established. Pretreatment of positive reference serum with the antigens of different bacteria demonstrated no decrease, whereas the serum pretreated with F. necrophorum antigens revealed a decrease in the ELISA values. The apparent difference in ELISA values was observed between the sera derived from cattle infected and not infected with this bacterium. The ELISA was also employed for the detection of the transition of immunoglobulins G and M in the infected cattle and revealed the increase of specific IgG in the animals. These findings indicated that the use of the ELISA for the immunoglobulin detection may prove to be a useful tool for predictive study of F. necrophorum infection in cattle.
In addition, partial purification and characterization of the HCl antigen was conducted. The antigen was partially purified by CM cellulose column chromatography and treatment of freeze-thawing. The partially purified preparation exhibited one definite and two faint bands in the lane of SDS-PAGE. Molecular weight of the former was estimated as 16 kDa.
|
Report
(4 results)
Research Products
(18 results)
