Rapid, automated nucleic acid probe assays using silicon microstructures for nucleic acid concentration
Rapid, automated nucleic acid probe assays using silicon microstructures for nucleic acid concentration
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DOI：
10.1115/1.2798037
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发表时间：
1999-02-01
影响因子：
1.7
通讯作者：
Northrup, MA
中科院分区：
文献类型：
--
作者：
Christel, LA;Petersen, K;Northrup, MA
描述了一种基于PCR技术的用于快速使用点核酸（NA）分析的系统。从测试样品中提取和浓缩DNA已经利用具有高表面积与体积比的硅流体微芯片完成。短（500 bp）和中等大小（48，000 bp）的DNA已被捕获、洗涤并使用这些芯片的二氧化硅表面洗脱。离液（GuHCl）盐溶液用作粘合剂。洗涤剂和洗脱剂分别由乙醇基溶液和水组成。在100-1000 ng/mL浓度范围内，从输入溶液中捕获接近40 ng/cm（2）结合面积的DNA量。对于用于病原体检测的感兴趣的稀释样品，使用PCR和凝胶电泳来证明约50%的提取效率，并且使用噬菌体λ DNA作为靶的约10倍的浓缩因子也已经证明了具有集成固态检测组件的快速多通道PCR热循环模块。这些结果证实了利用这些组件作为紧凑的一次性盒系统的元件的可行性，用于在诸如临床诊断、生物战剂检测、食品质量控制和环境监测的应用中检测NA。
A system for rapid point-of-use nucleic acid (NA) analysis based on PCR techniques is described. The extraction and concentration of DNA from test samples has been accomplished utilizing silicon fluidic microchips with high surface-area-to-volume ratios. Short (500 bp) and medium size (48,000 bp) DNA have been captured, washed, and eluted using the silicon dioxide surfaces of these chips. Chaotropic (GuHCl) salt solutions were used as binding agents. Wash and elution agents consisted of ethanol-based solutions and water, respectively. DNA quantities approaching 40 ng/cm(2) of binding area were captured from input solutions in the 100-1000 ng/mL concentration range. For dilute samples of interest for pathogen detection, PCR and gel electrophoresis were used to demonstrate extraction efficiencies of about 50 percent, and concentration factors of about 10x using bacteriophage lambda DNA as the target Rapid, multichannel PCR thermal cycling modules with integrated solid-state detection components have also been demonstrated. These results confirm the viability of utilizing these components as elements of a compact, disposable cartridge system for the detection of NA in applications such as clinical diagnostics, biowarfare agent detection, food quality control, and environmental monitoring.