이전|다음

신장이식 후 BK 바이러스 및 JC 바이러스 감염 검출을 위한 CRISPR 기반 검사

  • 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*

추상적인

장기 이식 수혜자는 이식편 생존을 위협하는 급성 또는 만성 감염으로 인한 상당한 위험에 직면합니다. BK 바이러스(BKV)와 JC 바이러스(JCV)는 두 가지 주요 기회 감염 바이러스로, 신장이식 후 면역 억제 상태에 있는 환자에서 폴리오마바이러스 관련 신병증과 이식 신장의 손실을 유발할 수 있습니다. 따라서 바이러스 부하의 적시 감지와 지속적인 모니터링이 필수적입니다. 그러나 현재 진단 방법은 제한적이며 새로운 분자 진단 기술 개발이 매우 시급합니다. BKV 및 JCV 검출을 위해 CRISPR RNA(crRNA) 서열 및 농도, Cas13a 농도, 그리고 재조합효소 중합효소 증폭(RPA) 프라이머가 최적화되었습니다. 다음으로, 새로운 마이크로플루이딕 이중 방울 칩이 설계 및 제작되었으며 CRISPR(ddCRISPR)와 통합되어 BKV와 JCV를 동시에 정성적으로 검출할 수 있습니다. 이후 ddCRISPR 검사는 임상 샘플을 이용해 검증되었습니다. 그리고 자원이 제한된 환경에서도 BKV와 JCV를 검출할 수 있는 CRISPR 결합 측면 유동 스트립(LFCRISPR)이 개발되었습니다. BKV 및 JCV 검출을 위해 원팟 RPA-CRISPR 반응 시스템이 구축되고 최적화되었습니다. ddCRISPR는 높은 민감도(BKV 10 복제본/ml, JCV 1 복제본/ml)로 BKV 및 JCV를 동시에 빠르게 검출할 수 있으며 절대 정량화가 가능하여 바이러스 부하 검출에 적합하고 장기 이식 수혜자의 맞춤형 정밀 치료에 유리합니다. LFCRISPR는 간단한 시각적 판독을 통해 작업 과정을 단순화하여 이식 후 바이러스 선별을 용이하게 합니다. 이러한 플랫폼은 분자 검사를 이식 치료 모델에 통합하여 비용을 절감하고 이식편 생존 시간을 연장하며 신장이식 후 수술 후 관리의 임상 결과를 개선하고 환자의 삶의 질을 향상시킵니다.

키워드

BK 바이러스 (BKV); JC 바이러스 (JCV); 신장 이식; 정기적으로 간격을 둔 짧은 회문 반복 서열 (CRISPR); 마이크로플루이딕; 측면 유동 검사 (LFA)

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