A fluorometric assay for rapid enrichment and determination of bacteria by using zirconium-metal organic frameworks as both capture surface and signal amplification tag
A fluorometric assay for rapid enrichment and determination of bacteria by using zirconium-metal organic frameworks as both capture surface and signal amplification tag
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使用锆金属有机框架作为捕获表面和信号放大标签来快速富集和测定细菌的荧光测定法
DOI：
10.1007/s00604-020-4136-8
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发表时间：
2020-02
影响因子：
5.7
通讯作者：
Xie Guoming
中科院分区：
文献类型：
--
作者：
Yang Shuangshuang;Guo Yongcan;Fan Jingchuan;Yang Yujun;Zuo Chen;Bai Shulian;Sheng Shangchun;Li Junjie;Xie Guoming
采用荧光法测定鲍曼不动杆菌(A。利用Zr-mMOF作为磁性fe3o4纳米颗粒的功能涂层提供修饰表面（Zr-mMOF），并作为荧光素载体产生荧光信号（F@UIO-66-NH2）。通过强Zr-O-P键合，两个不同的末端磷酸盐标记了eda。将鲍曼特异性适配体和脂多糖（LPS）特异性适配体分别附着在zr - mof上，制备磁核-壳捕获探针（标记为zr - mof -p- ab - apt）和信号探针（标记为F@UIO-66-NH2-p-LPS-Apt）。在a。鲍曼蛋白血样与磁分离，采用夹心式复合捕获探针/A。用高浓度的阴离子磷酸盐离子处理鲍曼细胞/信号探针，破坏信号探针中uio -66- nh2的纳米结构，快速释放荧光素，产生放大的荧光信号。由于zr - mof -p- ab - apt具有较高的适体修饰效率（高达93%）和对a的强亲和力。捕获探针对鲍曼尼虫的富集效率达到96.7%。此外，由于uio -66- nh2的高荧光素加载效率和我们的新型扩增策略破坏F@UIO-66-NH2-p-LPS-Apt在高浓度阴离子磷酸盐离子存在下在512 nm释放和放大荧光素信号，该方法的灵敏度达到10 cfu mL−1。该方法允许富集和测定a。a的检出限为~2.5 h。鲍曼球蛋白血样品浓度为10cfu mL - 1，线性范围为101-105cfu mL - 1。这表明该试验在早期血流感染诊断中的潜力。图示：采用捕获探针（zr - mof -p- ab - apt）和信号探针（F@UIO-66-NH2-p-LPS-Apt）对鲍曼不动杆菌血液样品进行三明治式荧光检测的示意图。检测限低至10 cfu mL-1，线性范围为101-105cfu mL-1。
A fluorometric assay was introduced to determineAcinetobacter baumannii(A.baumannii) in blood samples by utilizing Zr-MOFs both as functional coating for magnetic Fe3O4nanoparticles to provide modification surface (Zr-mMOF) and as fluorescein carrier to produce fluorescence signals (F@UIO-66-NH2). Through strong Zr-O-P bonding, two distinct terminal phosphate-labeledA.baumanniiand lipopolysaccharide (LPS) specific aptamers were attached onto Zr-MOFs to fabricate the magnetic core-shell capture probe (denoted as Zr-mMOF-p-Ab-Apt) and signal probe (denoted as F@UIO-66-NH2-p-LPS-Apt), respectively. After successive incubation withA.baumanniiin blood samples and magnetic separation, the sandwich-type composite of capture probe/A.baumanniicells/signal probe was treated with high concentration of anionic phosphate ions to destroy the nano-structure of UIO-66-NH2in the signal probe and fast release of fluorescein to produce amplified fluorescence signals. Due to the high aptamer modification efficiency of Zr-mMOF-p-Ab-Apt (up to 93%) and its strong affinity toA.baumannii, the enrichment efficiency of this capture probe has reached to 96.7%. Further, due to the high fluorescein loading efficiency of UIO-66-NH2and our novel amplification strategy to destroy F@UIO-66-NH2-p-LPS-Apt to release and amplify fluorescein signals at 512 nm in the presence of high concentration of anionic phosphate ions, the sensitivity of this method has reached 10 cfu mL−1. This method allows enrichment and determination ofA.baumanniiwithin ~2.5 h. The limit of detection ofA.baumanniiin blood samples is 10 cfu mL−1with a linear range of 101–105cfu mL−1. This indicates the potential of this assay for diagnosis of bloodstream infection in early stage.Graphical abstractSchematic representation of sandwich–type fluorometric assay forAcinetobacter baumanniiin blood samples with the capture probe (Zr-mMOF-p-Ab-Apt) and signal probe (F@UIO-66-NH2-p-LPS-Apt). The limit of detection is down to 10 cfu mL-1with a linear range of 101-105cfu mL-1.
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