基于 GMR 生物传感器阵列的 cDNA 定量分析用于即时检测基因表达分析。
Quantification of cDNA on GMR biosensor array towards point-of-care gene expression analysis.
机构信息
Department of Bioengineering, Stanford University, Stanford, CA 93405, USA.
Department of Materials Science and Engineering, Stanford University, Stanford, CA 93405, USA.
出版信息
Biosens Bioelectron. 2019 Apr 1;130:338-343. doi: 10.1016/j.bios.2018.09.050. Epub 2018 Sep 13.
Gene expression analysis at the point-of-care is important for rapid disease diagnosis, but traditional techniques are limited by multiplexing capabilities, bulky equipment, and cost. We present a gene expression analysis platform using a giant magnetoresistive (GMR) biosensor array, which allows multiplexed transcript detection and quantification through cost-effective magnetic detection. In this work, we have characterized the sensitivity, dynamic range, and quantification accuracy of Polymerase chain reaction (PCR)-amplified complementary DNA (cDNA) on the GMR for the reference gene GAPDH. A synthetic GAPDH single-stranded DNA (ssDNA) standard was used to calibrate the detection, and ssDNA dilutions were qPCR-amplified to obtain a standard curve. We demonstrate that the GMR platform provides a dynamic range of 4 orders of magnitude and a limit of detection of 1 pM and 0.1 pM respectively for 15 and 18-cycle amplified synthetic GAPDH PCR products. The quantitative results of GMR analysis of cell-line RNA were confirmed by qPCR.
即时医疗点的基因表达分析对于快速疾病诊断非常重要，但传统技术受到多重检测能力、设备庞大和成本的限制。我们提出了一种使用巨磁电阻（GMR）生物传感器阵列的基因表达分析平台，该平台通过经济高效的磁检测实现了多重转录本的检测和定量。在这项工作中，我们对 GMR 上经聚合酶链反应（PCR）扩增的互补 DNA（cDNA）的灵敏度、动态范围和定量准确性进行了表征，用于参考基因 GAPDH。使用合成的 GAPDH 单链 DNA（ssDNA）标准对检测进行了校准，并对 ssDNA 稀释液进行了 qPCR 扩增以获得标准曲线。我们证明，GMR 平台分别为 15 个和 18 个循环扩增的合成 GAPDH PCR 产物提供了 4 个数量级的动态范围和 1 pM 和 0.1 pM 的检测限。通过 qPCR 对细胞系 RNA 的 GMR 分析的定量结果进行了验证。
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