Establishment of highly sensitive ELISA to determine specific IgM antibody for diagnosis of acute cat scratch disease and elucidation of its pathology
| Project/Area Number |
17K09013
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Yamaguchi University |
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Principal Investigator |
TSUNEOKA Hidehiro 山口大学, 大学院医学系研究科, 教授(特命) (40437629)
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| Co-Investigator(Kenkyū-buntansha) |
大津山 賢一郎 山口大学, 大学院医学系研究科, 助教 (10432741)
徳田 信子 獨協医科大学, 医学部, 教授 (70227578)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 猫ひっかき病 / Bartonella henselae / ELISA-IgM / ELISA / ウエスタンブロット / IgM-WB / ウエスタンブロット法 / ＩｇＭ-ＥＬＩＳＡ |
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| Outline of Final Research Achievements |
This study aimed to establish a highly sensitive IgM ELISA using recombinant protein antigens of Bartonella henselae for serodiagnosis of cat scratch disease (CSD). We used seven types of candidate genus as antigens found at 8-10kDa of the major protein in the B. henselae IgM antibody-positive patients and examined the antigenicity of these proteins by two methods: cell-free protein synthesis and protein synthesis introduced into Escherichia coli.
As a result, genus protein expression was confirmed in all the genera in both methods, while responsiveness to CSD patients’ sera was not confirmed and no antigenicity was clarified in the Western blot assay. Further investigation will be required to establish the optimal IgM ELISA for the detection of CSD.
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| Academic Significance and Societal Importance of the Research Achievements |
猫ひっかき病（CSD）の病原菌Bartonella henselae は培養困難なため、血清学的診断法が有用であり高感度ELISAの開発が求められている。本研究は未だ存在しない“特異性の高い組換え蛋白質抗原を使用した高感度ELISA-IgM抗体価測定法の確立”を目指し、研究代表者らの一連の研究よりB. henselae菌体の8-10kDaの解析から研究を進めた。その結果、遺伝子解明とその組換え蛋白質生成合成はできたものの、それら蛋白質の抗原性は認められず、更なる検討が求められた。本ELISA確立時には一層のCSDの診断率向上と病態の解明が期待される。
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Report
(4 results)
Research Products
(6 results)
