| Project/Area Number |
17590486
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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 | University of Toyama |
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Principal Investigator |
KITAJIMA Isao Univ.of Toyama, Clin.Molecular Pathology, Professor, 大学院医学薬学研究部, 教授 (50214797)
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| Co-Investigator(Kenkyū-buntansha) |
KISHI Hiroyuki Univ.of Toyama, immunology, Associate Professor, 大学院医学薬学研究部, 助教授 (60186210)
YASUOKA Akira Univ.of Nagasaki, Clin.Infect Diseases, Professor, 医学部, 教授 (80242113)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | systemic inflammatory response syndrome (SIRS) / inflammatory cytokine / rapid testing / transcriptional factor / Nuclear factor-kappa B / sepsis / ELISA / surface plasmon resonance (SPR) / 1分子計測 |
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| Research Abstract |
The nuclear factor-kappa B (NF-κB) family of transcription factors is known to play an important role in the regulation of the immune system. NF-κB is activated by bacterial and viral antigens, which leads to the production of proinflammatory cytokines and chemokines. The rapid detection of activated NF-κB activation by the systemic inflammatory response syndrome (SIRS) is considered to be crucial for the treatment of patients with septicemia. The aim of the present study was to evaluate the sensitivity of two methods, electrophoretic mobility shift assay (EMSA) and transcription factor enzyme-linked immunoassay (TF-ELISA). TF-ELISA detected 25 ng of recombinant human NF-κB p50 (rhp50) and 5 mg of TNFα-stimulated HeLa nuclear protein, while EMSA detected approximately 100 ng of rhp50 and 10 mg of HeLa nuclear protein. We found that the TF-ELISA was more sensitive than EMSA in detecting NF-κB. However, it was judged that the 3-6 hour measuring time required in TF-ELISA was excessively long for patients with SIRS. Therefore, the development of new analytical methods with improved sensitivity and, measurement time was necessary for the detection and quantification of activated NF-κB protein in the hospital laboratory. Consequently, we have developed a new NF-κB analyzer based on surface plasmon resonance (SPR), which is recognized as one of the most sensitive direct optical detection methods. This method can detect nanomolar concentrations of NF-κB within 15 minutes. At present, we are carrying out clinical trials using this new transcription factor analysis apparatus for SIRS patients.
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