用于甘蔗黄叶病毒飞摩尔检测的磁性纳米酶增强型快速免疫流式检测法

IF 2.3 Q1 AGRICULTURE, MULTIDISCIPLINARY ACS agricultural science & technology Pub Date : 2024-06-28 DOI:10.1021/acsagscitech.4c00274
Raja Muthuramalingam Thangavelu*, Janiga PK, Nithyanantham Ramasamy and Viswanathan Rasappa, 
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引用次数: 0

摘要

植物病害严重威胁全球粮食安全,历史上曾多次发生过毁灭性的流行病。这种风险在甘蔗等主要农作物和粮食作物中尤为严重。尽管分子诊断技术的最新进展提高了甘蔗病毒的检测水平,但这些方法由于依赖于复杂、昂贵的设备,在很大程度上仅限于实验室环境。为了克服这一局限性,我们开发了一种更容易获得且成本效益更高的解决方案:磁性纳米酶增强比色免疫流式检测法,专为超灵敏检测甘蔗黄叶卷曲病毒(ScYLV)而设计。这项创新技术可对低至飞摩尔水平的病毒浓度进行清晰的光学鉴定。该检测方法采用阳离子磁性纳米粒子进行病毒分离,并使用比色免疫标记进行诊断,从而提高了灵敏度,并可提供即时结果,其灵敏度可与定量实时聚合酶链反应(qRT-PCR)和酶联免疫吸附测定(ELISA)等成熟方法相媲美。我们的检测方法提供一步检测过程和两步半定量分析,标志着植物病毒诊断领域的重大突破。我们对检测方法的设计进行了广泛的研究,包括传感平台、阻断剂、抗体连接化学、灵敏度、定量、多路复用潜力和现场适用性。这项诊断研究在流式分析法中使用了磁性酶,是植物病害诊断中快速灵敏诊断的开创性方法。它为传统的分子诊断引入了一种前景广阔的替代方法,有可能改变植物病害管理,提高全球粮食安全。
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Magnetic Nanozyme-Enhanced Rapid ImmunoFlow-Through Assay for the Femtomolar Detection of Sugarcane Yellow Leaf Virus

Plant diseases significantly threaten global food security, with numerous historical instances of devastating epidemics. This risk is particularly acute in key agricultural and food crops, such as sugarcane. Although recent advancements in molecular diagnostics have improved the detection of sugarcane viruses, these methods are largely confined to lab settings due to their reliance on sophisticated, costly equipment. To overcome this limitation, we have developed a more accessible and cost-effective solution: a magnetic nanozyme-enhanced colorimetric ImmunoFlow-through assay designed for the ultrasensitive detection of sugarcane yellow leaf curl virus (ScYLV). This innovative technique allows for clear optical identification of viral concentrations as low as femtomolar levels. The assay employs cationic magnetic nanoparticles for virus isolation and colorimetric immunolabels for diagnosis, enhancing sensitivity and providing immediate results, comparable to those of established methods like quantitative real-time-polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Our assay offers a one-step detection process and a two-step semiquantitative analysis, marking a major breakthrough in plant virus diagnostics. Extensive research into the assay’s design, including its sensing platform, blocking agents, antibody conjugation chemistries, sensitivity, quantification, potential for multiplexing, and field applicability, was carried out. This diagnostic research utilizing Magnetozyme in a flow-through assay represents a pioneering approach to rapid and sensitive diagnosis within plant disease diagnostics. It introduces a promising alternative to traditional molecular diagnostics, potentially transforming plant disease management and enhancing food security globally.

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