Anal Chem. 2011 Feb 1;83(3):690-5. doi: 10.1021/ac102858j. Epub 2010 Dec 10.
Multiplex gene assay is a valuable molecular tool not only in academic science but also in clinical diagnostics. Multiplex PCR assays, DNA microarrays, and various nanotechnology-based methods are examples of major techniques developed for analyzing multiple genes; none of these, however, are suitable for point-of-care diagnostics, especially in resource-limited settings. In this report, we describe an octopus-like multiplex microfluidic loop-mediated isothermal amplification (mμLAMP) assay for the rapid analysis of multiple genes in the point-of-care format and provide a robust approach for predicting viruses. This assay with the ability of analyzing multiple genes qualitatively and quantitatively is highly specific, operationally simple, and cost/time-effective with the detection limit of less than 10 copies/μL in 2 μL quantities of sample within 0.5 h. We successfully developed a mμLAMP chip for differentiating three human influenza A substrains and identifying eight important swine viruses.
多重基因检测是一种非常有价值的分子工具，不仅在学术科学中，而且在临床诊断中也是如此。多重 PCR 检测、DNA 微阵列和各种基于纳米技术的方法是分析多个基因的主要技术示例；然而，这些方法都不适合即时诊断，特别是在资源有限的环境中。在本报告中，我们描述了一种章鱼样的多重微流控环介导等温扩增（mμLAMP）检测方法，用于即时诊断格式中快速分析多个基因，并提供了一种强大的病毒预测方法。该方法具有定性和定量分析多个基因的能力，具有高度特异性、操作简单、成本/时间效益高，检测限在 0.5 小时内，2 μL 样本量中低于 10 拷贝/μL。我们成功开发了一种 mμLAMP 芯片，用于区分三种人类流感 A 亚株，并鉴定出八种重要的猪病毒。
