Simultaneous DNA amplification and detection using a pH-sensing semiconductor system
Simultaneous DNA amplification and detection using a pH-sensing semiconductor system
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DOI：
10.1038/nmeth.2520
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发表时间：
2013-07-01
期刊：
影响因子：
48
通讯作者：
Zhang, Linglan
中科院分区：
文献类型：
--
作者：
Toumazou, Christofer;Shepherd, Leila M.;Zhang, Linglan
我们利用互补金属氧化物半导体（CMOS）技术开发了一种用于核酸实时扩增和检测的集成芯片。在这里，我们展示了一种放大耦合检测方法，用于直接测量核苷酸掺入过程中释放的氢离子，而不是依赖于间接测量，如荧光染料。这是一种无标记、非光学、实时方法，用于通过监测天然扩增化学的pH特征来检测和定量靶序列。该芯片集成了离子敏感场效应晶体管（ISFET）传感器、温度传感器、电阻加热、信号处理和控制电路，可创建完整的片上系统平台。我们使用两种扩增策略：PCR和等温扩增来评估该平台。使用这个平台，我们基因分型和歧视独特的单核苷酸多态性（SNP）变异的细胞色素P450家族从原油人类唾液。我们预计这种半导体技术将能够创造出具有成本效益、便携式和可扩展的实时核酸分析设备。
We developed an integrated chip for real-time amplification and detection of nucleic acid using pH-sensing complementary metal-oxide semiconductor (CMOS) technology. Here we show an amplification-coupled detection method for directly measuring released hydrogen ions during nucleotide incorporation rather than relying on indirect measurements such as fluorescent dyes. This is a label-free, non-optical, real-time method for detecting and quantifying target sequences by monitoring pH signatures of native amplification chemistries. The chip has ion-sensitive field effect transistor (ISFET) sensors, temperature sensors, resistive heating, signal processing and control circuitry all integrated to create a full system-on-chip platform. We evaluated the platform using two amplification strategies: PCR and isothermal amplification. Using this platform, we genotyped and discriminated unique single-nucleotide polymorphism (SNP) variants of the cytochrome P450 family from crude human saliva. We anticipate this semiconductor technology will enable the creation of devices for cost-effective, portable and scalable real-time nucleic acid analysis.