Li Yuanyuan, Xu Fei, Zhang Jinhua, Huang Jinshan, Shen Di, Ma Yunmiao, Wang Xiufeng, Bian Yuan, Chen Qing
Department of Orthopaedics, Zhuji Affiliated Hospital of Shaoxing University, Zhuji 311800, Zhejiang, China.
Department of Urology, Taizhou Hospital of Zhejiang Province, Linhai 317000, China.
ACS Omega. 2021 Apr 30;6(18):12223-12228. doi: 10.1021/acsomega.1c01107. eCollection 2021 May 11.
Rapid and sensitive pathogenic bacterial identification and isolation from complicated clinical specimens are of great importance for the early diagnosis and prevention of osteomyelitis. Herein, we proposed a novel methicillin-resistant (MRSA) detection strategy through two specially designed streptavidin magnetic bead-based probes, including a capture probe and a report probe. In detail, the capture probe takes the responsibility to specially bind with the surface protein of MRSA and leads to the liberation of the promoter which could subsequently initiate report probe-based signal amplification. Afterward, the hybridization of the promoter probe with the report probe could then transform the protruding 3' terminus of template DNA in the report probe into a blunt end. With the assistance of Exo III, the template could be digested to liberate the promoter to form a recycle and to liberate the biprobe to induce the following rolling circle amplification (RCA)-based signal amplification. Through the integration of the Exo III-assisted recycle and RCA-based signal amplification, the proposed method exhibited a favorable detection performance.
从复杂临床标本中快速、灵敏地鉴定和分离致病细菌对于骨髓炎的早期诊断和预防至关重要。在此，我们通过两种特殊设计的基于链霉亲和素磁珠的探针，提出了一种新型耐甲氧西林金黄色葡萄球菌（MRSA）检测策略，包括捕获探针和报告探针。具体而言，捕获探针负责与MRSA的表面蛋白特异性结合，并导致启动子的释放，随后启动基于报告探针的信号放大。之后，启动子探针与报告探针的杂交可将报告探针中模板DNA的突出3'末端转化为平端。在Exo III的协助下，模板可被消化以释放启动子，形成循环，并释放双探针以诱导随后基于滚环扩增（RCA）的信号放大。通过整合Exo III辅助循环和基于RCA的信号放大，该方法表现出良好的检测性能。
