Microfluidic chip for multiple detection of miRNA biomarkers in breast cancer based on three-segment hybridization
Microfluidic chip for multiple detection of miRNA biomarkers in breast cancer based on three-segment hybridization
复制标题
基于三段杂交的乳腺癌miRNA生物标志物多重检测微流控芯片
DOI：
10.1063/1.5137784
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发表时间：
2020-04-01
期刊：
影响因子：
1.6
通讯作者：
Han, Lin
中科院分区：
文献类型：
--
作者：
Gao, Yakun;Qiang, Le;Han, Lin
迫切需要建立一个快速、方便、准确、低成本的miRNA定量检测平台，这对疾病的发展和癌症的早期诊断具有重要意义。在这里，我们提出了一个针对miRNA的检测微流控平台，将自组装的聚L赖氨酸底物与微流控芯片集成在一起，基于三段杂交技术同时对多个样本中的miRNA进行多个检测。PLL首先自组装到干净的玻璃片上，然后与高通量微印刷微流控芯片集成在一起，以局部动员DNA探针。设计了一种加载样品的微流控芯片，实现了多个样品的同时多个检测。三段杂交系统用于检测miRNAs，其中捕获探针与目标miRNA的一端互补，带有荧光的检测探针与靶miRNA的另一端互补。首先，将捕获探针固定在芯片上，并将具有荧光的检测探针与目标miRNA杂交。其次，miRNA检测探针杂交器与固定在芯片上的捕获探针反应。最后，清洗多余的检测探针，用激光扫描仪检测芯片上捕获探针-miRNA-检测探针杂交物的荧光强度。选择4个乳腺癌标志物miRNAs同时检测，检测下限为1 pm，检测时间为30分钟。该微流控平台对多种样本中miRNAs的检测具有较高的灵敏度，有望用于乳腺癌的早期诊断。
It is urgent to establish a fast, convenient, accurate, and low-cost miRNA quantitative detection platform, which is important in disease development and the early diagnosis of cancer. Here, we propose a miRNA-specific detection microfluidic platform in which a self-assembled Poly-L-Lysine (PLL) substrate is integrated with microfluidic chips and conduct multiple detection of miRNAs from multiple samples at the same time based on three-segment hybridization. PLL is first self-assembled onto a clean glass slide and then integrated with a high-throughput micro-printing microfluidic chip to locally mobilize DNA probes. A sample-loading microfluidic chip is designed to realize multiple detection of multiple samples at the same time. A three-segment hybridization system is used to detect miRNAs in which the capture probe is complementary to one end of the target miRNA and the detection probe with fluorescence is complementary to the other end of the target miRNA. First, capture probes are mobilized on the chip and detection probes with fluorescence are hybridized with the target miRNA. Second, a miRNA-detection probe hybridizer is reacted with the capture probes immobilized on the chip. Finally, excessive detection probes are cleaned and the fluorescence intensity of the capture probe–miRNA–detection probe hybridizer on the chip is detected by using a laser scanner. Four significant breast cancer biomarker miRNAs are selected for simultaneous detection, and the detection limit is 1 pM with a detection time of 30 min. This microfluidic platform shows sensitive multiple detection of miRNAs in multiple samples and is promising for the early diagnosis of breast cancer.