Smart pH-Regulated Switchable Nanoprobes for Photoelectrochemical Multiplex Detection of Antibiotic Resistance Genes
Smart pH-Regulated Switchable Nanoprobes for Photoelectrochemical Multiplex Detection of Antibiotic Resistance Genes
复制标题
用于抗生素耐药基因光电化学多重检测的智能 pH 调节可切换纳米探针
DOI：
10.1021/acs.analchem.0c02839
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发表时间：
2020-08-18
影响因子：
7.4
通讯作者：
Gong, Jingming
中科院分区：
文献类型：
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作者：
Li, Xin;Lu, Junmiao;Gong, Jingming
由特定基因编码的抗生素耐药性已成为全球主要的健康威胁和医疗保健的重大负担。因此，对抗生素耐药基因(ARGs)的简便、低成本、准确的检测在人类健康和安全领域是至关重要的，尤其是多重传感检测。本文建立了一种灵巧的pH调节可切换光电化学(PEC)生物分析方法，用于超灵敏地检测青霉素耐药基因的两种典型亚型：bla(-ctx-M-1)(靶标1，标记为T-DNA 1)和bla(-tee)(靶标2，标记为T-DNA 2)。PEC生物测定的操作依赖于pH响应型S-DNA-Sbt@SiO_2 NSS络合物在外界pH刺激下的可切换解离，从而启动预先嵌入夹心型DNA纳米组装的离子的pH调节释放。在酸性条件下，SDNAI标签的解离(开启状态)触发嵌入的SBO+的释放。在碱性条件下，S-DNA标签的解离被抑制(关闭状态)。T-DNA2的检测是通过DNA杂交触发的金属离子释放实现的。引入的发夹T-Hg2+-T片段的解旋与第二个锚定信号DNA(S-DNA 2)杂交，点燃了Hg2+的释放。释放出的SbO+或Hg2+离子通过与光敏层的离子交换，触发Sb2S3/ZnS或HgS/ZnS异质结的形成，产生放大的光电流，最终实现对青霉素抗性基因bla(CTX-M-1)和bla(-TEM)的超灵敏检测。构建的pH调节PEC生物检测方法巧妙地集成了pH响应智能单元如S-DNA标签、pH调节嵌入离子的释放以及随后的基于离子交换的信号放大策略，显示出高灵敏度、特异性、低成本和易于使用的多重检测ARGs。它可以成功地用于测量真实大肠杆菌质粒中的bla(-CTX-M-1)和bla(-TEM)，展示了开发新型遗传护理点设备的巨大前景。
Antibiotic resistance, encoded via particular genes, has become a major global health threat and substantial burden on healthcare. Hence, the facile, low-cost, and precise detection of antibiotic resistance genes (ARGs) is crucial in the realm of human health and safety, especially multiplex sensing assays. Here, a smart pH-regulated switchable photoelectrochemical (PEC) bioassay has been created for ultrasensitive detection of two typical subtypes of penicillin resistance genes bla(-CTX-M-1) (target 1, labeled as T-DNA1) and bla(-TEE) (target 2, labeled as T-DNA2), whereby pH-responsive antimony tartrate (SbT) complex-grafted silica nanospheres are ingeniously adopted as signal DNA1 tags (labeled as S-DNA1-SbT@SiO(2)NSs). The operations of the PEC bioassay depend on the switchable dissociation of the pH-responsive S-DNA1-SbT@SiO2 NSs complex under the external pH stimuli, thus initiating the pH-regulated release of ions pre-embedded in sandwich-type DNA nanoassemblies. At acidic conditions, the dissociation of SDNAI tags (ON state) triggers the release of the embedded SbO+. Under alkaline conditions, the dissociation of S-DNA1 tags is inhibited (OFF state). The detection of T-DNA2 was achieved via DNA hybridization-triggered metal ion release. The unwinding of the introduced hairpin T-Hg2+-T fragment, hybridized with the second anchored signal DNA (S-DNA2), ignites the release of Hg2+. The released SbO+ or Hg2+ ions would trigger the formation of Sb2S3/ZnS or HgS/ZnS heterostructure through ion-exchange with the photosensitive ZnS layer, giving rise to the amplified photocurrents and eventually realizing the ultrasensitive detection of penicillin resistance genes subtypes, bla(CTX-M-1) and bla(-TEM). The as-fabricated pH-regulated PEC bioassay, smartly integrating the pH-responsive intelligent unit as S-DNA tags, pH-regulated release of embedded ions, and the subsequent ion-exchange-based signal amplification strategy, exhibits high sensitivity, specificity, low-cost, and ease of use for multiplex detection of ARGs. It can be successfully used for measuring bla(-CTX-M-1) and bla(-TEM) in real E. coli plasmids, demonstrating great promise for developing a new class of genetic point-of-care devices.