A PCR reactor with an integrated alumina membrane for nucleic acid isolation
A PCR reactor with an integrated alumina membrane for nucleic acid isolation
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DOI：
10.1039/c0an00288g
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发表时间：
2010-01-01
期刊：
影响因子：
4.2
通讯作者：
Bau, Haim H.
中科院分区：
文献类型：
--
作者：
Kim, Jitae;Mauk, Michael;Bau, Haim H.
最近，人们对能够检测临床和环境样品中的核酸（NA）的即时检测装置越来越感兴趣。然而，核酸检测需要使装置操作复杂化的各种样品制备步骤。一种有吸引力的补救措施是将许多（如果不是全部）样品制备操作和核酸扩增集成到单个反应室中。描述了一种微流控芯片，其在单个腔室中将聚合酶链式反应（PCR）扩增与使用纳米多孔氧化铝膜（AOM）的核酸固相提取集成。将疑似含有目标细菌和/或病毒的样品与裂解剂和离液盐混合，并加载到容纳纳米多孔氧化铝膜的塑料芯片中。裂解物中的核酸与膜结合。然后洗涤膜，用PCR反应试剂填充腔室，并且循环腔室的温度以扩增捕获的核酸并产生可检测的产物。DNA和RNA（逆转录）的分离和扩增证明。由于干膜对液体流动的高阻力，设计了专门的流量控制系统以促进样品引入和膜洗涤。
Recently, there has been a growing interest in point-of-care devices capable of detecting nucleic acids (NA) in clinical and environmental samples. Nucleic acid detection requires, however, various sample preparation steps that complicate device operation. An attractive remedy is to integrate many, if not all, sample preparation operations and nucleic acid amplification into a single reaction chamber. A microfluidic chip that integrates, in a single chamber, polymerase chain reaction (PCR) amplification with solid-phase extraction of nucleic acids using a nanoporous, aluminium oxide membrane (AOM) is described. Samples suspected of containing target bacteria and/or viruses are mixed with lysis agents and a chaotropic salt and loaded into a plastic chip housing a nanoporous, aluminium oxide membrane. The nucleic acids in the lysate bind to the membrane. The membrane is then washed, the chamber is filled with the PCR reaction reagents, and the chamber's temperature is cycled to amplify the captured nucleic acids and produce detectable products. Both DNA and RNA (with reverse-transcription) isolation and amplification are demonstrated. Due to the dry membrane's high resistance to liquid flow, a specialized flow control system was devised to facilitate sample introduction and membrane washing.