2001 Fiscal Year Final Research Report Summary
Development of assays detecting antibodies to parasite proteinases for immunodiagnosis of parasitic diseases
| Project/Area Number |
11670255
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
寄生虫学(含医用動物学)
|
|---|
| Research Institution | Kanazawa Medical University |
|---|
Principal Investigator |
IKEDA Teruaki Kanazawa Medical University, Medical Zoology, Associate Professor, 医学部, 助教授 (70064607)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
OIKAWA Yosaburo Kanazawa Medical University, Medical Zoology, Research Assistant, 医学部, 助手 (10139785)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Keywords | paragonimiasis / fascioliasis / immunodiagnosis / cysteine proteinase / cystatin / Protein A / IgE / immunocapture |
|---|
| Research Abstract |
Enzyme-linked immunosorbent assays (ELISAs) which detect specific antibodies to fluke cysteine proteinases have provided good sensitivity and specificity for immunodiagnosis of trematode diseases. To detect the specific antibodies without the need for purified proteinase antigens, three immunocapture assays using cystatin (protein inhibitor of cysteine proteinase), Protein A, or anti-human IgE monoclonal antibody were applied for the immunodiagnosis of paragonimiasis and fascioliasis. In a cystatin capture ELISA, cystatin-sensitized plate wells were incubated with a preparation containing fluke cysteine proteinases and with diluted patient sera, and standard ELISA procedures were then followed. In Protein A and anti human IgE antibody capture assays, sensitized wells were incubated with diluted patient sera, the fluke proteinase preparation, then fluorogenic peptidyl substrate for fluke cysteine proteinases. These two assays detected specific IgG and IgE, respectively, to fluke cysteine proteinases by measuring proteinase activity on the plate wells. The three immunocapture assays, which detected IgG and IgE specific for fluke cysteine proteinases, provided a high sensitivity and specificity for immunodiagnosis of paragonimiasis and fascioliasis. The detection of specific IgE is superior to that of specific IgG in evaluating the efficacy of chemotherapy.
|
