Decision Applications Division, Mail Stop K551, Los Alamos National Laboratory, Los Alamos, NM 87545, United States.
J Microbiol Methods. 2010 Feb;80(2):155-63. doi: 10.1016/j.mimet.2009.12.001. Epub 2009 Dec 16.
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.
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。我们称之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检测方法不同，我们的方案由单个管反应组成，然后杂交到 Luminex 微球阵列进行检测。我们证明了该检测方法能够在单个多重反应中同时检测多种核酸特征（例如，独特序列、单核苷酸多态性）。检测探针由模块化组件组成，能够实现目标检测、探针扩增，以及随后的微球阵列捕获。为了证明我们的检测方法的实用性，我们将其应用于检测三种生物威胁剂，炭疽杆菌、鼠疫耶尔森菌和土拉弗朗西斯菌。结合该检测方法的简便性和稳健性，这里呈现的结果表明我们的检测方法在诊断和监测方面具有很大的应用潜力。
