Utility of fully automated real-time PCR for the rapid diagnosis of implant infection

• Project/Area Number: 18K16669
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 56020:Orthopedics-related
• Research Institution: Yokohama City University
• Principal Investigator: CHOE Hyonmin 横浜市立大学, 医学部, 講師 (20760888)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥130,000 (Direct Cost: ¥100,000、Indirect Cost: ¥30,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 全自動リアルタイムPCR / メチシリン耐性遺伝子 / 16sRNA / 遺伝子診断 / インプラント周囲感染 / 人工関節周囲感染 / 整形外科感染 / 骨軟部組織感染症 / MRS感染 / 関節液 / 全自動リアルタイムPCRの精度の検証 / リアルタイムPCR / 細菌性DNA / 全自動

Outline of Final Research Achievements

Fully automated real-time PCR: GENE QUBE was used to evaluate its usefulness for rapid diagnosis of periprosthetic infection. Fully automatic real-time PCR included MRS-PCR targeting methicillin resistance genes and 16sRNA-PCR targeting 16sRNA genes possessed by all bacteria. They were used for the joint fluid and tissues collected from patients with periprosthetic infection. We demonstrated that the fully automatic MRS-PCR and 16sRNA-PCR provides rapid and accurate diagnosis compared with microbiological culture result. As a new finding, uncertain substances in synovial fluid and tissues has possibility to inhibit the PCR reaction and is possibly responsible to the false negative PCR results.

Academic Significance and Societal Importance of the Research Achievements

全自動PCRは迅速性と精度にすぐれ、遺伝子検査にて懸念される偽陽性の可能性も提言することが期待できる。全自動PCRを用いた細菌感染の診断は、整形外科感染症領域における診断・治療において患者へ負担提言が、治療の適正化、耐性菌における医療負担の低減といった面での良い効果が期待できる

Research Products

• [Journal Article] Detection of mecA and 16S rRNA Genes Using Real-Time PCR Can Be Useful in Diagnosing Iliopsoas Abscess, Especially in Culture-Negative Cases: RT-PCR for Iliopsoas Abscess (2022)
  • Author(s): Hyonmin Choe, Naomi Kobayashi, Yohei Ito, Hiroyuki Ike, Taro Tezuka, Masanobu Takeyama, Yusuke Kawabata, Yutaka Inaba
  • Journal Title: Biomed Research International Volume: なし Pages: 1-7
  • DOI: 10.1155/2022/2209609
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] An automated real‐time PCR assay for synovial fluid improves the preoperative etiological diagnosis of periprosthetic joint infection and septic arthritis (2020)
  • Author(s): Yang Fan、Choe Hyonmin、Kobayashi Naomi、Tezuka Taro、Oba Masatoshi、Miyamae Yushi、Morita Akira、Abe Koki、Inaba Yutaka
  • Journal Title: Journal of Orthopaedic Research Volume: 39 Issue: 2 Pages: 348-355
  • DOI: 10.1002/jor.24959
  • Peer Reviewed / Int'l Joint Research
• [Presentation] PJIにおけるBiofilmの重要性 (2021)
  • Author(s): 崔賢民, 小林直実, 稲葉裕.
  • Organizer: 日本整形外科学会総会, シンポジウム
• [Presentation] 関節液に対する全自動リアルタイムPCRを用いたメチシリン耐性ブドウ球菌感染の術前診断 (2021)
  • Author(s): 崔賢民, 池裕之, 手塚太郎, 稲葉裕.
  • Organizer: 日本整形外科学会総会, シンポジウム
• [Presentation] 整形外科感染の診断における関節液中の細菌DNA検出の有用性 (2020)
  • Author(s): 崔賢民
  • Organizer: 日本整形外科基礎学術集会