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基于碳点作为光学纳米探针的比率荧光共振能量转移适体传感器用于尿液样本中鲍曼不动杆菌的高灵敏和选择性检测

Ratiometric fluorescence resonance energy transfer aptasensor for highly sensitive and selective detection of Acinetobacter baumannii bacteria in urine sample using carbon dots as optical nanoprobes.

作者信息

Bahari Delnia, Babamiri Bahareh, Salimi Abdollah, Salimizand Himen

机构信息

Department of Chemistry, University of Kurdistan, 66177-15175, Sanandaj, Iran.

Department of Chemistry, University of Kurdistan, 66177-15175, Sanandaj, Iran; Research Center for Nanotechnology, University of Kurdistan, 66177-15175, Sanandaj, Iran.

出版信息

Talanta. 2021 Jan 1;221:121619. doi: 10.1016/j.talanta.2020.121619. Epub 2020 Sep 6.

Abstract

Development of sensitive and selective analytical method for accurate diagnosis of Acinetobacter baumannii (Ab) bacteria in biological samples is a challenge. Herein, we developed an ingenious ratiometric fluorescent aptasensor for sensitive and selective detection of (Ab) bacteria based on fluorescence resonance energy transfer (FRET) between ortho-phenylenediamines carbon dot (o-CD), nitrogen-doped carbon nanodots (NCND) as donor's species and graphene oxide (GO) as acceptor. NCND that assembled onto the edge of graphene oxide (GO) exhibited quenched photoluminescence emission, and with the absorption of the modified o-CD with aptamer (o-CD-ssDNA) onto the graphene oxide surface the fluorescence of o-CD was efficiently quenched. The aptamer (ssDNA) as a biorecognition element is bound with A. baumannii specifically which releases the o-CD-ssDNA from GO and the recovery of the fluorescence signal of o-CD, while the fluorescence intensity of NCND only slightly altered and acted as the reference signal in ratiometric fluorescence assay. The fluorescence intensity ratio (I nm/Inm) varied from 2.0 to 10.0 with the concentration of bacteria changing from 2.0 × 10 to 4.5 × 10 cfu/mL and the low detection limit of 3.0 × 10 cfu/mL (S/N = 3). The feasibility of the developed aptasensor for selective detection of A. baumannii in urine sample with satisfactory results was also demonstrated.

摘要

开发用于准确诊断生物样本中鲍曼不动杆菌(Ab)细菌的灵敏且具选择性的分析方法是一项挑战。在此,我们基于邻苯二胺碳点(o-CD)、氮掺杂碳纳米点(NCND)作为供体物种与氧化石墨烯(GO)作为受体之间的荧光共振能量转移(FRET),开发了一种用于灵敏且选择性检测(Ab)细菌的巧妙比率荧光适配体传感器。组装在氧化石墨烯(GO)边缘的NCND表现出光致发光发射猝灭,随着用适配体修饰的o-CD(o-CD-ssDNA)吸附到氧化石墨烯表面,o-CD的荧光被有效猝灭。作为生物识别元件的适配体(ssDNA)与鲍曼不动杆菌特异性结合,从而使o-CD-ssDNA从GO上释放出来,o-CD的荧光信号得以恢复,而NCND的荧光强度仅略有变化,并在比率荧光测定中作为参考信号。随着细菌浓度从2.0×10变为4.5×10 cfu/mL,荧光强度比(I nm/Inm)从2.0变化到10.0,检测下限低至3.0×10 cfu/mL(S/N = 3)。还证明了所开发的适配体传感器用于尿液样本中鲍曼不动杆菌选择性检测的可行性,结果令人满意。

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