| Project/Area Number |
06672300
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Jichi Medical School (JMS) |
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Principal Investigator |
OBAYASHI Taminori Jichi Medical School, Dept.of Medicine, Assistant Professor, 医学部, 講師 (60102266)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAI Tadashi Jichi Medical School, Dept.of Medicine, Professor, 医学部, 教授 (60048957)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
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| Keywords | deep mycosis / (1*3) -beta-D-glucan / triple helix / single strand / automation / horseshoe crab / (1*3) -beta-D-glucan binding protein / Factor G / カブトガニ血球抽出液 / 抗(1-3)-β-D-グルカン抗体 / ELISA / 三重鎖螺旋構造 / カイネティック・モード |
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| Research Abstract |
Automatic assay of plasma (1*3) -beta-D-glucan with limulus reagent has been materialized with a new pretreating mixture which obviates the necessity of centrifugation. The mixture consists of KOH,KCL,and polybrene. KOH is to remove limulus-test interfering factors and enhance the sensitivity of the test by dissembling the triple helix of the glucan into single strands, which are a more potent activator of factor G.KCL is to keep the dissembled glucan in single strands, and polybrene is to prevent the formation of nonspecific turbidity. The adoption of kinetic mode has broadened the dynamic range up to 500 pg/ml. With a cutoff of 20 pg/ml, the sensitivity and the specificity of the test for invasive deep mycoses were 90% and 100%, respectively.
Recently a new protein was isolated from horseshoe crab hemolymph that specifically binds to (1*3) -beta-D-glucan, thereby inhibiting the activation of factor G by the glucan. Using this new protein and anti- (1*3) -beta-D-glucan antibody, we developed an ELISA for (1*3) -beta-D-glucan determination. Plasma glucan concentrations in invasive deep mycoses showed a good correlation between the two methods, another evidence that the glucan is increased in blood in this conditon.
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