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腎移植後のBKウイルスおよびJCウイルス感染検出のためのCRISPRベースのアッセイ

  • role： Contribute equally, First author同等贡献者, 第一作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Yu Liu1,  
  • role： Contribute equally同等贡献者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Jing-Song Xu1,  
  • role： Contribute equally同等贡献者
  • Affiliation：

    Department of Pathology, the Fifth People’s Hospital of Shanghai, Fudan University, Shanghai 200240, China

Li Cao2,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Shuang Yang1,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Tian-Ming Li1,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Hai-Qian Huang1,  
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Jun-Heng Zhang1,  
  • Affiliation：

    Central Laboratory, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

Xue Zhao3,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

  • Email：qq2005011@163.com
  • Introduction：* Qian Liu qq2005011@163.com
Qian Liu1*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China

  • Email：lishun86@126.com
  • Introduction：Shun Li lishun86@126.com
Shun Li4*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

  • Email：rjlimin@shsmu.edu.cn
  • Introduction：Min Li rjlimin@shsmu.edu.cn
Min Li1*,  
  • role： Corresponding author通信作者
  • Affiliation：

    Department of Laboratory Medicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China

  • Email：renjijywh@shsmu.edu.cn
  • Introduction：Hua Wang renjijywh@shsmu.edu.cn
Hua Wang1*

サマリー

臓器移植の受容者は、移植片の生存を脅かす急性または慢性の感染による重大なリスクに直面しています。BKウイルス（BKV）とJCウイルス（JCV）は二つの主要な日和見感染ウイルスであり、腎移植後に免疫抑制状態にある患者に多腫瘍ウイルス関連腎症および移植腎の喪失を引き起こす可能性があります。したがって、ウイルス量の適時検出および継続的なモニタリングが不可欠です。しかし、現在の診断方法は依然として制限があり、新しい分子検出技術の開発は非常に緊急を要します。CRISPR RNA（crRNA）配列および濃度、Cas13aの濃度、および組換えポリメラーゼ増幅（RPA）のプライマーは、BKVおよびJCV検出のために最適化されました。次に、新しいマイクロ流体二重滴チップが設計および製造され、CRISPR（ddCRISPR）と統合されてBKVおよびJCVを同時に定性的に検出しました。その後、ddCRISPRアッセイは臨床試料を用いて検証されました。次に、資源が限られた環境でのBKVおよびJCV検出のために、CRISPRと組み合わせた側流試験紙（LFCRISPR）が開発されました。BKVおよびJCV検出のためにワンポットRPA-CRISPR反応システムが確立および最適化されました。ddCRISPRは高感度（BKVは10コピー/ml、JCVは1コピー/ml）でBKVおよびJCVを同時かつ迅速に検出でき、絶対定量を提供し、ウイルス量の検出に適しており、臓器移植受容者の個別化および精密治療に役立ちます。LFCRISPRは簡単な視覚的読み取りにより操作プロセスを簡素化し、臓器移植後のウイルススクリーニングを促進します。これらのプラットフォームは分子検査を移植治療モデルに組み込み、コストを削減し、移植片の生存期間を延ばし、腎移植の術後管理における臨床成績を改善し、患者の生活の質を向上させます。

キーワード

BKウイルス（BKV）；JCウイルス（JCV）；腎移植；クラスター規則的に間隔をあけた短い回文配列（CRISPR）；マイクロ流体；側流アッセイ（LFA）

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