Establishment of rapidly identification of unknown pathogenic microorganisms using melting temperature mapping method in bone joint soft tissue infections

2021 Fiscal Year Final Research Report

• Project/Area Number: 18K09043
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56020:Orthopedics-related
• Research Institution: Kitasato University
• Principal Investigator: Uchiyama Katsufumi 北里大学, 医学部, 教授 (90286310)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Keywords: 人工関節周囲感染 / 感染 / PCR / Tm mapping 法 / 16S-rRNA / 関節液 / 骨関節軟部組織感染症 / 菌数定量的検査

Outline of Final Research Achievements

The melting temperature mapping method is a new rapid identification system for the causative agent of septicemia developed by Dr. Niimi et al.
In this study, the melting temperature mapping method was used for rapid diagnosis of bone joint soft tissue infection. In order to examine the reliability of tests using infected soft tissue other than joint fluid and to establish a diagnostic method for multiple bacterial infections, we confirmed multiple unknown pathogenic microorganisms using a next-generation sequencer.
We also evaluated the usefulness of the quantitative test of "bacterial count" in the diagnosis of periprosthetic joint infection, and examined the cut-off value of the bacterial count.

Free Research Field

整形外科

Academic Significance and Societal Importance of the Research Achievements

Tm mapping 法ではプライマー数を７つに固定して広範な菌種を同定できる。データベースの修正・拡張が容易であり、新たなmutant strainが出現しても直ぐに登録して、同定に反映させることが出来る。塩基配列を読むことなく、PCRとmelting解析のみで同定できるため、迅速・簡便・安価な同定が可能であり、耐性菌の診断や菌数定量が可能であることから治療効果の判定に応用できる可能性がある。