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一种用于核酸提取和检测的集成生物传感器平台。

An integrated biosensor platform for extraction and detection of nucleic acids.

机构信息

CNR-IMM, Catania, Italy.

STMicroelectronics, Catania, Italy.

出版信息

Biotechnol Bioeng. 2020 May;117(5):1554-1561. doi: 10.1002/bit.27290. Epub 2020 Feb 5.

Abstract

The development of portable systems for analysis of nucleic acids (NAs) is crucial for the evolution of biosensing in the context of future healthcare technologies. The integration of NA extraction, purification, and detection modules, properly actuated by microfluidics technologies, is a key point for the development of portable diagnostic systems. In this paper, we describe an integrated biosensor platform based on a silicon-plastic hybrid lab-on-disk technology capable of managing NA extraction, purification, and detection processes in an integrated format. The sample preparation process is performed by solid-phase extraction technology using magnetic beads on a plastic disk, while detection is done through quantitative real-time polymerase chain reaction (qRT-PCR) on a miniaturized silicon device. The movement of sample and reagents is actuated by a centrifugal force induced by a disk actuator instrument. The assessment of the NA extraction and detection performance has been carried out by using hepatitis B virus (HBV) DNA genome as a biological target. The quantification of the qRT-PCR chip in the hybrid disk showed an improvement in sensitivity with respect to the qRT-PCR commercial platforms, which means an optimization of time and cost. Limit of detection and limit of quantification values of about 8 cps/reaction and 26 cps/reaction, respectively, were found by using analytical samples (synthetic clone), while the results with real samples (serum with spiked HBV genome) indicate that the system performs as well as the standard methods.

摘要

便携式核酸(NA)分析系统的发展对于未来医疗技术中的生物传感发展至关重要。通过微流控技术适当驱动的 NA 提取、纯化和检测模块的集成,是开发便携式诊断系统的关键点。在本文中,我们描述了一种基于硅塑料混合芯片实验室技术的集成生物传感器平台,该平台能够以集成格式管理 NA 提取、纯化和检测过程。样品制备过程通过塑料盘上的磁性珠固相萃取技术进行,而检测则通过微型硅设备上的定量实时聚合酶链反应(qRT-PCR)进行。样品和试剂的移动由盘式执行器仪器产生的离心力驱动。通过使用乙型肝炎病毒(HBV)DNA 基因组作为生物靶标,评估了 NA 提取和检测性能。在混合磁盘中的 qRT-PCR 芯片的定量分析表明,与商用 qRT-PCR 平台相比,灵敏度有所提高,这意味着时间和成本的优化。使用分析样品(合成克隆)时,检测限和定量限分别约为 8 cps/反应和 26 cps/反应,而使用真实样品(含有 HBV 基因组的血清)的结果表明该系统与标准方法一样有效。

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