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Highly sensitive visual detection of Avian Influenza A (H7N9) virus based on the enzyme-induced metallization

机译:基于酶诱导的金属化的禽流感A(H7N9)病毒的高灵敏视觉检测

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摘要

Development of convenient but sensitive method for influenza detection is highly important in immediate and effective clinical treatment. In this study, an ultrasensitive colorimetric approach combining the advantages of the convenience of the enzyme-induced metallization and the high specificity of enzyme-linked immunosorbent assay for the detection of influenza virus A (H7N9 as model) has been developed. Two rounds of amplification are utilized to enhance the detection sensitivity. The amplification of enzymatic reaction combines with the specific optical properties of gold nanoparticles causing the enhancing of the optical signal immensely. In addition, the increased surface area and the magnetic enrichment effect also enable the magnetic bead (MB) to catch a large number of alkaline phosphatase (ALP) and detection antibody (Ab(2)), thus very small amounts of the virus can be easily detected. Compared with conventional method, this approach exhibits outstanding sensitivity for ALP detection, 0.2 U/L of ALP can be distinguished with a spectrometer and 2 U/L with the naked eye. And as low as 25 pg/mL of H7N9 can be detected by the naked eye. This approach shows an extensive horizon for bioassays and is available in clinical diagnosis with the advances of simplification, effectiveness, low cost and sensitive readout. (C) 2016 Elsevier B.V. All rights reserved.
机译:开发便捷而灵敏的流感检测方法对于立即有效的临床治疗至关重要。在这项研究中,已经开发了一种结合酶诱导的金属化便利性和酶联免疫吸附法的高特异性检测流感病毒A(H7N9作为模型)的优点的超灵敏比色方法。利用两轮扩增来增强检测灵敏度。酶促反应的放大与金纳米颗粒的特定光学性质相结合,极大地增强了光信号。此外,增加的表面积和磁富集效应还使磁珠(MB)能够捕获大量的碱性磷酸酶(ALP)和检测抗体(Ab(2)),因此可以携带非常少量的病毒容易检测到。与常规方法相比,该方法对ALP检测具有出色的灵敏度,可以用分光光度计区分0.2 U / L的ALP和用肉眼区分2 U / L。肉眼可检测到低至25 pg / mL的H7N9。这种方法显示了生物测定的广阔前景,并且随着简化,有效性,低成本和灵敏读数的发展,可用于临床诊断。 (C)2016 Elsevier B.V.保留所有权利。

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