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16766

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A fluorometric assay for rapid enrichment and determination of bacteria by using zirconium-metal organic frameworks as both capture surface and signal amplification tag.

著者 Yang S , Guo Y , Fan J , Yang Y , Zuo C , Bai S , Sheng S , Li J , Xie G
Mikrochim Acta.2020 Feb 24 ; 187(3):188.
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A fluorometric assay was introduced to determine Acinetobacter baumannii (A. baumannii) in blood samples by utilizing Zr-MOFs both as functional coating for magnetic FeO nanoparticles to provide modification surface (Zr-mMOF) and as fluorescein carrier to produce fluorescence signals (F@UIO-66-NH). Through strong Zr-O-P bonding, two distinct terminal phosphate-labeled A. baumannii and 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-NH-p-LPS-Apt), respectively. After successive incubation with A. baumannii in blood samples and magnetic separation, the sandwich-type composite of capture probe/A. baumannii cells/signal probe was treated with high concentration of anionic phosphate ions to destroy the nano-structure of UIO-66-NH in 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 to A. baumannii, the enrichment efficiency of this capture probe has reached to 96.7%. Further, due to the high fluorescein loading efficiency of UIO-66-NH and our novel amplification strategy to destroy F@UIO-66-NH-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. This method allows enrichment and determination of A. baumannii within ~2.5 h. The limit of detection of A. baumannii in blood samples is 10 cfu mL with a linear range of 10-10 cfu mL. This indicates the potential of this assay for diagnosis of bloodstream infection in early stage. Graphical abstractSchematic representation of sandwich-type fluorometric assay for Acinetobacter baumannii in 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 with a linear range of 10-10 cfu mL.
PMID: 32095939 [PubMed - in process]
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