A rapid multiplex assay for nucleic acid-based diagnostics
A rapid multiplex assay for nucleic acid-based diagnostics
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DOI：
10.1016/j.mimet.2009.12.001
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发表时间：
2010-02-01
影响因子：
2.2
通讯作者：
White, P. Scott
中科院分区：
文献类型：
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作者：
Deshpande, Alina;Gans, Jason;White, P. Scott
我们开发了一种快速（4 小时内）、多重核酸检测方法，适用于微球阵列检测平台。我们将此测定称为多重寡核苷酸连接 PCR (MOL-PCR)。与其他需要多个步骤的基于连接的测定不同，我们的方案包括单管反应，然后与 Luminex 微球阵列杂交进行检测。我们证明了该测定在单个多重反应中同时检测不同核酸特征（例如，独特序列、单核苷酸多态性）的能力。检测探针由模块化组件组成，可实现目标检测、探针扩增以及随后捕获到微球阵列上。为了证明我们的检测方法的实用性，我们将其应用于三种生物威胁因子的检测：炭疽芽孢杆菌、鼠疫耶尔森菌和土拉热杆菌。结合该测定的简便性和稳健性，此处提供的结果表明我们的测定在诊断和监测中具有强大的潜力。由 Elsevier B.V. 出版
We have developed a rapid (under 4 hours), multiplex, nucleic acid assay, adapted to a microsphere array detection platform. We call this assay multiplex oligonucleotide ligation-PCR (MOL-PCR). Unlike other ligation-based assays that require multiple steps, our protocol consists of a single tube reaction, followed by hybridization to a Luminex microsphere array for detection. We demonstrate the ability of this assay to simultaneously detect diverse nucleic acid signatures (e.g., unique sequences, single nucleotide polymorphisms) in a single multiplex reaction. Detection probes consist of modular components that enable target detection, probe amplification, and subsequent capture onto microsphere arrays. To demonstrate the utility of our assay, we applied it to the detection of three biothreat agents, B. anthracis, Y. pestis, and F. tularensis. Combined with the ease and robustness of this assay, the results presented here show a strong potential of our assay for use in diagnostics and surveillance. Published by Elsevier B.V.