Develoment of MRSA rapid detection method using phage derived ligand protein

• Project/Area Number: 15K21360
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine; Infectious disease medicine
• Research Institution: Kitasato University
• Principal Investigator: Matsui Hidehito 北里大学, 北里生命科学研究所, 研究員 (80503797)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: MRSA / ファージ / 迅速診断 / 診断 / イムノクロマト

Outline of Final Research Achievements

Bacteriophages infect with a narrow range of host cells by binding on the specific cell-surface receptor. This high host-specificity is largely relies on the specific interaction of the ligand protein and the receptor molecule. This project aimed to develop a simple and rapid Staphylococcus aureus diagnosis test employing the phage ligand protein in the lateral flow assay. The ligand protein was expressed by the gene cloning technique and purified by the affinity chromatography. The test was able to detect 32 clinical isolates of S. aureus tested without exception and showed no cross reactivity with other microorganisms including 31 bacterial species and one strain of yeast so far tested. We previously developed a MRSA detection tool using a monoclonal antibody raised against PBP2’ in the immunochromatogaphy. The combination of the present method and the PBP2’ detection tool enabled to distinguish MRSA and MSSA.

Research Products

• [Presentation] ラテラルフローアッセイによるMRSA検出方法の開発 (2017)
  • Author(s): 松井秀仁, 津田愛美, 内山淳平, 花木秀明
  • Organizer: MRSAフォーラム2017
• [Presentation] PBP2’イムノクロマトによるOS-MRSAの検出 (2016)
  • Author(s): 松井秀仁, 津田愛美, 花木秀明
  • Organizer: 第86回日本感染症学会 西日本地方会学術集会
  • Place of Presentation: 沖縄コンベンションセンター(沖縄県, 宜野湾市）
  • Year and Date: 2016-11-25
• [Presentation] Development of a phage protein-mediated Staphylococcus aureus detection tool (2016)
  • Author(s): Hidehito Matsui, Jumpei Uchiyama, Megumi Tsuda, Shigenobu Matsuzaki, Taiji Nakae and Hideaki Hanaki
  • Organizer: The 116th General Meeting American Sociaty of Microbiology
  • Place of Presentation: Boston Convention and Exhibition Center(Boston, Massachusetts, USA)
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] イムノクロマト法を用いたメチシリン耐性コアグラーゼ陰性ブドウ球菌の検出 (2016)
  • Author(s): 松井秀仁、津田愛美、花木秀明
  • Organizer: 第64回日本化学療法学会総会
  • Place of Presentation: 神戸国際会議場(兵庫県, 神戸市)
  • Year and Date: 2016-06-10
• [Presentation] ファージを利用した細菌検出 ～黄色ブドウ球菌検出ファージクロマトの開発～ (2016)
  • Author(s): 松井秀仁, 内山淳平, 津田愛美, 松崎茂展, 花木秀明
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪国際交流センター(大阪府大阪市)
  • Year and Date: 2016-03-25
  • Invited
• [Presentation] MRSA感染症と 迅速診断方法の開発 (2016)
  • Author(s): 松井秀仁
  • Organizer: 第61回防衛衛生学会 臨床検査技師研修会 三宿地区
  • Place of Presentation: 陸上自衛隊三宿駐屯地(東京都世田谷区)
  • Year and Date: 2016-02-03
  • Invited
• [Presentation] 細菌感染症における診断法と治療法の開発：バクテリオファージの利用 (2015)
  • Author(s): 内山淳平、松井秀仁、阪口義彦、花木秀明、松崎茂展、阪口雅弘
  • Organizer: 第158回日本獣医学会学術集会
  • Place of Presentation: 北里大学獣医学部(青森県十和田市)
  • Year and Date: 2015-09-08
  • Invited
• [Presentation] ファージ尾部タンパクを用いた黄色ブドウ球菌検出方法の開発 (2015)
  • Author(s): 松井秀仁、崔龍洙、花木秀明
  • Organizer: 第63回日本化学療法学会総会
  • Place of Presentation: 京王プラザホテル(東京都新宿区)
  • Year and Date: 2015-06-06