| Co-Investigator(Kenkyū-buntansha) |
YOKOSUKA Osamu Chiba University, Medical School, assistant, 医学部, 助手 (90182691)
TAGAWA Masami Chiba University, Medical School, research fellow, 医学部, 医員
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1988: ¥900,000 (Direct Cost: ¥900,000)
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| Research Abstract |
We have tried to identify the enteric non-A, non-B hepatitis virus and their proteins by western blotting, immune-electron microscopy, and genetic engineering techniques.
1 . Fecal materials from patients with enteric non-A, non-B hepatitis were filtered and the filtrants were ultracentrifuged. Then both supernatents and pellets were electrophoresed through 10% acridamide gel. By protein staining with Coomassie blue, bands were observed at size of 100, 64, and 26 kilodalton. However, no specific bands were identified by western blotting using patients' serum at their convalescent stage.
2. By light microscopic examination, histological findings of enteric non-A, non-B hepatitis were almost identical with those of other acute hepatitis. By electron microscopic examination, virus-like particles were observed in the cyteplasm of degenerated hepatocytes. However, we could not identify them as enteric non-A, non-B hepatitis virus particles by immune-electron microscopic examination.
3. Ribonucleic acids (RNA) were extracted from patients fecal materials, then reverse transcribed to cDNA, and ligated with lambda gt 11 vector. The lambda gt 11 library were screened with patients convalescent phase serum, but no positive clone was obtained. Although we examined these RNA by polymerase chain reaction methods, no positive findings were obtained as well.
The reason why we could not obtain the positive finding may be because either the materials were not good; enough to get viral RNA or because antibody production is too weak to detect viral proteins. Further examination is now underway to identify the virus nucleic acids and viral proteins with fecal materials and convalescent phase sera from other patients.
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