2012 Fiscal Year Final Research Report
Tecnological development of endotoxin assay with leukocyte-rich plasma and plasma in sepsis
| Project/Area Number |
22390339
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Emergency medicine
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| Research Institution | Iwate Medical University |
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Principal Investigator |
ENDO Shigeatsu 岩手医科大学, 医学部救急医学講座, 教授 (30160394)
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| Co-Investigator(Renkei-kenkyūsha) |
SUZUKI Yasushi 岩手医科大学, 医学部救急医学, 講師 (90306019)
KOJIKA Masahiro 岩手医科大学, 医学部救急医学, 助教 (40347878)
KIKKAWA Tomohiro 岩手医科大学, 医学部救急医学, 助教 (90468314)
TAKAHASHI Gaku 岩手医科大学, 医学部救急医学, 助教 (60453304)
SHIBATA Shigehiro 岩手医科大学, 医学部救急医学, 助教 (10326671)
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| Project Period (FY) |
2010 – 2012
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| Keywords | 集中治療医学 / 敗血症 |
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| Research Abstract |
For the National Health Insurance-covered endotoxin assay method for the determination of plasma endotoxin, namely, turbidimetry-gelation time assay, platelet-rich plasma (PRP) prepared by centrifugation of the patient ’s blood is used, as a rule. However, the diagnostic sensitivity of this assay can hardly be considered to be satisfactory (sensitivity, 58.6%; specificity, 97.0%). In sepsis, endotoxin recognized by leukocytes and bound to the surface of leukocytes as a bacterial cell constituent and endotoxin liberated from bacteria and bound to the CD14 receptor, toll-like receptor (TLR) 4, etc., on the leukocyte membrane surface are known; in addition endotoxin is also considered to be taken up by the cells besides being present in the circulating plasma. Endotoxin thereby activates leukocytes, which in turn, is associated with activation of humoral factors, such as cytokines. We focused our attention on the endotoxin occurring within and on the surface of white blood cells, and have reported the possibility of the sensitivity of the test being improved by simultaneously using both white blood cells and plasma as assay samples. This study was undertaken to devise and examine a new method of sample preparation for endotoxin assay in leukocyte-rich plasma (LRP), prepared taking advantage of the property of hydroxyethyl starch (HES) as an erythrocyte aggregating agent. Furthermore, we comparatively assessed the assay results obtained by the conventional method using PRP and the turbidimetry-gelation time method using LRP. The results showed significantly higher endotoxin levels in LRP, as compared to the endotoxin levels in PRP determined by the conventional method, in patients with sepsis, and a significantly higher sensitivity of the former method. We consider that our newly devised assay method may contribute to improvement of the diagnosis rate of sepsis.
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