2021 Fiscal Year Final Research Report

Construction of a comprehensive sepsis testing system based on the new sepsis definition and its operational deployment in Japan and Southeast Asia.

Research Project

PDF

Project/Area Number	18H02728
Research Category	Grant-in-Aid for Scientific Research (B)
Allocation Type	Single-year Grants
Section	一般
Review Section	Basic Section 52010:General internal medicine-related
Research Institution	University of Toyama
Principal Investigator	Kitajima Isao 富山大学, 大学本部, 理事・副学長 (50214797)
Co-Investigator(Kenkyū-buntansha)	原田健右富山大学, 学術研究部医学系, 特命講師 (30436795) 柳原克紀長崎大学, 医歯薬学総合研究科(医学系), 教授 (40315239) 仁井見英樹富山大学, 学術研究部医学系, 准教授 (50401865) 山本善裕富山大学, 学術研究部医学系, 教授 (70452844)
Project Period (FY)	2018-04-01 – 2022-03-31
Keywords	敗血症 / 迅速細菌検査 / Tmマッピング法 / 敗血症起因菌定量検査 / 敗血症バイオマーカー / ATP測定 / NF-κB
Outline of Final Research Achievements	Vacuum blood collection tubes without cellular DNA contamination were prepared to establish an aseptic bacterial test using an automated nucleic acid extractor. Then, we developed a test method for determining blood bacterial load using real-time qPCR. The comprehensive septicemia testing system was established by combining the aforementioned test method with the Tm-mapping method developed in-house. A total of 1,166 clinical samples collected from adult patients with infectious diseases were evaluated to assess diagnostic accuracy and high rapidity of our test system. Samples collected from 156 pediatric patients with infectious diseases (256 samples) were also evaluated using our test system, which demonstrated its advantage in requiring less amount of blood (<1 mL) for evaluation. Further, evaluation of our test system of blood bacterial load revealed that the most acutely decreasing effective biomarker at 24 h after antibiotic therapy was "bacterial count in the blood."
Free Research Field	臨床検査
Academic Significance and Societal Importance of the Research Achievements	感染症起炎菌同定は迅速性が重要課題である。血液培養が標準検査であるが、2～3日を要している。Tmマッピング法は培養行程が省け、PCR産物の融解解析により、起炎菌が血液から直接同定が可能となり、世界最速レベルで起炎菌を提供できる。また、現時点で血液中の菌数を定量する検査方法は存在しないが、我々は、細菌DNAコピーから検体中の細菌数を算出できる検査法を開発した。敗血症起炎菌迅速同定・定量検査（菌名＆菌数/mL）の実用化にむけた臨床研究を実施し、その有用性を明らかにすることができた。敗血症迅速起炎菌同定と菌数を感染症重症度や治療効果の新たな指標として活用することは、敗血症治療への貢献が期待される