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通过 DNA zyme 催化扩增和原位聚集诱导发射光增敏剂生成的同步敏感免疫测定法及活 Zika 病毒的有效灭活。

Synchronously Sensitive Immunoassay and Efficient Inactivation of Living Zika Virus via DNAzyme Catalytic Amplification and In Situ Aggregation-Induced Emission Photosensitizer Generation.

机构信息

School of Public Health, Suzhou Medical College of Soochow University, Soochow University, Suzhou 215123, China.

Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.

出版信息

Anal Chem. 2024 Jun 4;96(22):9244-9253. doi: 10.1021/acs.analchem.4c01500. Epub 2024 May 21.

Abstract

Sensitive identification and effective inactivation of the virus are paramount for the early diagnosis and treatment of viral infections to prevent the risk of secondary transmission of viruses in the environment. Herein, we developed a novel two-step fluorescence immunoassay using antibody/streptavidin dual-labeled polystyrene nanobeads and biotin-labeled G-quadruplex/hemin DNAzymes with peroxidase-mimicking activity for sensitive quantitation and efficient inactivation of living Zika virus (ZIKV). The dual-labeled nanobeads can specifically bind ZIKV through E protein targeting and simultaneously accumulate DNAzymes, leading to the catalytic oxidation of Amplex Red indicators and generation of intensified aggregation-induced emission fluorescence signals, with a detection limit down to 66.3 PFU/mL and 100% accuracy. Furthermore, robust reactive oxygen species generated in situ by oxidized Amplex Red upon irradiation can completely kill the virus. This sensitive and efficient detection-inactivation integrated system will expand the viral diagnostic tools and reduce the risk of virus transmission in the environment.

摘要

敏感识别和有效失活病毒对于病毒感染的早期诊断和治疗至关重要,可防止病毒在环境中二次传播的风险。在此,我们开发了一种新颖的两步荧光免疫分析方法,使用抗体/链霉亲和素双重标记聚苯乙烯纳米珠和具有过氧化物酶模拟活性的生物素标记 G-四链体/血红素 DNA 酶,用于灵敏定量和有效灭活活 Zika 病毒(ZIKV)。双标记纳米珠可以通过靶向 E 蛋白特异性结合 ZIKV,同时聚集 DNA 酶,导致 Amplex Red 指示剂的催化氧化和产生增强的聚集诱导发射荧光信号,检测限低至 66.3 PFU/mL,准确率为 100%。此外,辐照时氧化 Amplex Red 原位产生的强活性氧可以完全杀死病毒。这种灵敏高效的检测-灭活集成系统将扩展病毒诊断工具,并降低环境中病毒传播的风险。

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