数字聚合酶链反应
核酸
微流控
DNA
数字化
核糖核酸
环介导等温扩增
核酸定量
注意事项
炸薯条
微流控芯片
化学
计算生物学
生物系统
纳米技术
材料科学
计算机科学
生物
聚合酶链反应
生物化学
基因
医学
病理
电信
计算机视觉
作者
Jiasi Wang,Jason E. Kreutz,Daniel T. Chiu
出处
期刊:Methods in molecular biology
日期:2021-11-26
卷期号:: 279-295
被引量:1
标识
DOI:10.1007/978-1-0716-1803-5_15
摘要
Digital nucleic acid quantitation methods show excellent sensitivity and specificity for pathogen detection. Droplet digital PCR (ddPCR) is the most advanced digital nucleic acid quantitation method and has been commercialized, but is not suitable for many point-of-care applications due to its complex instrumentation. Here we describe a simple microfluidics-based self-digitization (SD) chip for quantifying nucleic acids at the point of care with minimal instrumentation. We demonstrate the clinical diagnostic capability of this platform by applying it to quantifying human viral DNA and RNA. SD chips with a range of well numbers and volumes are tested, and isothermal methods are used to amplify the DNA and RNA to a detectable level. Sample concentration is determined based on the measured volume in the wells and the number of wells with fluorescence greater than a threshold based on a Poisson distribution. Concentration measurements over the low concentration range of 0-100 molecules/μL showed a strong correlation (R2 = 0.99) with measurements using a real-time PCR assay, demonstrating the sensitivity and specificity of the SD chip platform.
科研通智能强力驱动
Strongly Powered by AbleSci AI