Simultaneous Determination of Human Enterovirus 71 and Coxsackievirus B3 by Dual-Color Quantum Dots and Homogeneous Immunoassay
Simultaneous Determination of Human Enterovirus 71 and Coxsackievirus B3 by Dual-Color Quantum Dots and Homogeneous Immunoassay
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双色量子点均相免疫分析同时测定人肠道病毒71型和柯萨奇病毒B3型
DOI：
10.1021/ac203172x
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发表时间：
2012-04-03
影响因子：
7.4
通讯作者：
Wang, Hanzhong
中科院分区：
文献类型：
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作者：
Chen, Lu;Zhang, Xiaowei;Wang, Hanzhong
人类肠道病毒71型（EV71）和柯萨奇病毒B3型（CVB3）具有很高的发病率和死亡率。提出了一种采用两个彩色量子点（QDs）、病毒抗体和氧化石墨烯（GO）的病毒定量免疫测定方法。量子点为链亲和素共轭量子点（SA-QDs），抗体为生物素化抗体。生物素化EV71抗体（Ab1）通过生物素链亲和素相互作用与525 nm的绿色SA-QDs结合形成QDs-Ab1，而生物素化CVB3抗体（Ab2）通过生物素链亲和素相互作用与605 nm的红色SA-QDs结合形成QDs-Ab2。氧化石墨烯对QDs-Ab1和QDs-Ab2的荧光均有较好的猝灭作用。EV71和CVB3的靶点可以破坏QDs-Ab和GO复合物，分别恢复QDs-Ab1和QDs-Ab2的荧光。利用添加目标EV71和CVB3后两种不同颜色的QDs-Ab荧光恢复强度，可以在单一激发光下同时定量测定两种肠道病毒。按3倍信噪比计算，EV71和CVB3的检出限分别为0.42和0.39 ng mL（-1）。更重要的是，该策略可以进一步作为一种通用的方法，通过改变偶联抗体在疾病早期诊断中进行任何蛋白质或病毒的测定，为病毒的鉴别和测定提供了一种快速而有前景的临床方法。总之，建立了一种简便、快速、灵敏、高选择性的EV71和CVB3检测方法。该方法可用于临床样品分析，结果令人满意。值得注意的是，该传感器不仅可以通过荧光强度实现快速、精确的蛋白/病毒定量测定，还可以通过数字可视化技术实现半定量蛋白/病毒测定。
Human Enterovirus 71 (EV71) and Coxsackievirus B3 (CVB3) have high risks for morbidity and mortality. A virus quantitation immunoassay has been proposed by employing two colored quantum dots (QDs), antibodies of the virus, and graphene oxide (GO). The QDs are streptavidin-conjugated quantum dots (SA-QDs), and the antibodies are biotinylated antibodies. Biotinylated EV71 antibody (Ab1) was associated with 525 nm green colored SA-QDs via biotinstreptavidin interaction forming QDs-Ab1, whereas biotinylated CVB3 antibody (Ab2) was associated with 605 nm red colored SA-QDs via biotin streptavidin interaction forming QDs-Ab2. GO was an excellent quencher to the fluorescence of both QDs-Ab1 and QDs-Ab2. The targets of EV71 and CVB3 can break up the complex of QDs-Ab and GO, recovering the fluorescence of QDs-Ab1 and QDs-Ab2, respectively. Using these two different colored QDs-Ab fluorescence recovery intensities upon the addition of targets EV71 and CVB3, the two enteroviruses can be simultaneously quantitatively determined with a single excitation light. The detection limits of EV71 and CVB3 are 0.42 and 0.39 ng mL(-1) based on 3 times signal-to-noise ratio, respectively. More importantly, this strategy can be further used as a universal method for any protein or virus determination by changing the conjugated antibodies in disease early diagnosis, which can provide a fast and promising clinical approach for virus differentiation and determination. In a word, a simple, fast, sensitive, and highly selective assay for EV71 and CVB3 has been developed. It could be applied in clinical sample analysis with a satisfactory result. It was notable that the sensor could not only achieve rapid and precise quantitative determination of protein/virus by fluorescent intensity but also could be applied in semiquantitative protein/virus determination by digital visualization.