| Project/Area Number |
04670733
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General surgery
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
YOKOYAMA Takashi Hiroshima University, Medical Hospital, Professor, 医学部・附属病院, 教授 (60034104)
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| Co-Investigator(Kenkyū-buntansha) |
HIYAMA Eiko Hiroshima University, Medical Hospital, Research associate, 医学部・附属病院, 助手 (00218744)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Mec A / MRSA / Staphylococcal enterotoxin / TSST-1 / Molecular biology / Polymerase chain reaction / Cytokine / Tumor necrosis factor / PCR / TNFα / DNA / enterotoxin / mecA / 黄色ブドウ球菌 / 遺伝子診断 |
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| Research Abstract |
1. We have established the method for detection of genes for staphylococcal enterotoxins and TSST-1 by the polymelase chain reaction (PCR). Five pairs (ent A - D and tst) of oligonucleotid primers were made and tested with clinical isolated strains of S.aureus. The results using PCR and immunological method are almost same within ent A, B, D and tst. But ent C genes were detected by PCR with the strain of S.aureus whose enterotoxin C were not detected by immunological method. The toxical type of most causative strains of post operative MRSA enteritis was ent A, C and tst.
2. Next, PCR for mec A and tst was performed with directly extracted staphylococcal DNA from clinical samples (sputum and gastric juice). The sensibilitys for the detection of minimal CFU of MRSA were about 800 CFU for mec A and about 200 CFU for tst.
3. New Zealand white Rabbits were injected with TSST-1. Their arterial blood pressure were monitored and their white blood cell count was examined. The decline of the arterial blood pressure and decrease of white blood cell count were observed. But we could not detect serum TNF and IL-1 with bioassay.
4. We examined TNF production from human isolated monocyte (Mo) with stimulation of TSST-1 or LPS under various conditions (Mo, Mo + Tcell, Mo + IL-2, Mo + INF gamma, mono nuclear cells). There was no significant deferenciation of TNF level between each culture supernatant.
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