A novel fluorescence biosensor based on double-stranded DNA branch migration-induced HCR and DNAzyme feedback circuit for sensitive detection of Pseudomonas aeruginosa (clean version).
A novel fluorescence biosensor based on double-stranded DNA branch migration-induced HCR and DNAzyme feedback circuit for sensitive detection of Pseudomonas aeruginosa (clean version).
复制标题
DOI：
10.1016/j.aca.2022.340449
复制
发表时间：
2022-09
影响因子：
6.2
通讯作者：
Yaxing Xie;G. Xie;Jinshan Yuan;Jianhong Zhang;Yujun Yang;Yuan Yao;You Wu;Dan Bai;Kena Chen;Baiying Li;Lin Song;Hui Chen
中科院分区：
文献类型：
--
作者：
Yaxing Xie;G. Xie;Jinshan Yuan;Jianhong Zhang;Yujun Yang;Yuan Yao;You Wu;Dan Bai;Kena Chen;Baiying Li;Lin Song;Hui Chen
铜绿假单胞菌（P. aeruginosa）是医院感染中最常见的细菌之一。本文基于双链DNA分支迁移诱导杂交链反应（HCR）和DNA酶反馈电路，构建了一种新型的荧光生物传感器，用于敏感检测P.铜绿。P. aerodiosa与其适配体在DNA三向连接结构上引发双链DNA分支迁移，形成两个DNA“Y”连接结构。一个DNA“Y”连接结构打开荧光标记的DNA发夹并触发HCR。另一个DNA“Y”连接结构形成双链DNA酶并切割特异性核糖核苷酸位点，产生新的触发探针以开始双链DNA分支迁移的下一个循环。最终产生大量DNA“Y”型连接结构，极大地促进了信号放大。在优化的条件下，该生物传感器的检测线性范围为102- 107 CFU mL−1，检测限为37 CFU mL−1（S/N = 3）。回收率测试结果表明，该生物传感器具有良好的临床应用前景。由于HCR和DNAzyme通过双链DNA分支迁移同时启动反馈回路，所构建的生物传感器为病原菌检测提供了一个理想的平台，无需蛋白酶和复杂的信号放大过程。
Pseudomonasaeruginosa(P. aeruginosa) is one of the most common bacteria in nosocomial infection. Here, a novel fluorescence biosensor based on double-stranded DNA branch migration-induced hybridization chain reaction (HCR) and DNAzyme feedback circuit was constructed for sensitive detection ofP. aeruginosa. The binding ofP. aeruginosawith its aptamer on a DNA three-way junction structure initiated the double-stranded DNA branch migration to form two DNA “Y” junction structures. One DNA “Y” junction structure opened the fluorescence-labelled DNA hairpins and triggered the HCR. The other DNA “Y” junction structure formed a double-stranded DNAzyme and cleaved the specific ribonucleotide site, producing new triggering probes to start the next cycle of the double-stranded DNA branch migration. Ultimately, a large number of DNA “Y” junction structures were produced, which greatly promoted signal amplification. Under optimized conditions, the proposed biosensor detected a wide linearity range of 102–107CFU mL−1, and the limit of detection was 37 CFU mL−1(S/N = 3). The recovery test results indicated that the biosensor has promising clinical application potential. Because of the simultaneous initiation of the HCR and the DNAzyme feedback circuit through the double-stranded DNA branch migration, the constructed biosensor provided an ideal platform for pathogenic bacteria detection without protein enzymes and complex signal amplification procedures.