数字聚合酶链反应
环介导等温扩增
重组酶聚合酶扩增
消耗品
微尺度化学
多路复用
计算机科学
微流控
材料科学
计算机硬件
聚合酶链反应
纳米技术
DNA
生物芯片
化学
生物信息学
生物
基因
数学
数学教育
物理化学
生物化学
作者
Lei Cao,Xiaojin Guo,Ping Mao,Yulin Ren,Zedong Li,Minli You,Jie Hu,Miao Tian,Yao Chen,Fēi Li,Feng Xu
出处
期刊:ACS Sensors
[American Chemical Society]
日期:2021-10-04
卷期号:6 (10): 3564-3574
被引量:33
标识
DOI:10.1021/acssensors.1c00603
摘要
Digital polymerase chain reaction (dPCR) has found widespread applications in molecular diagnosis of various diseases owing to its sensitive single-molecule detection capability. However, the existing dPCR platforms rely on the auxiliary procedure to disperse DNA samples, which needs complicated operation, expensive apparatus, and consumables. Besides, the complex and costly dPCR readers also impede the applications of dPCR for point-of-care testing (POCT). Herein, we developed a portable digital loop-mediated isothermal amplification (dLAMP) platform, integrating a microscale hydrogel (microgel) array chip for sample partition, a miniaturized heater for DNA amplification, and a hand-held reader for digital readout. In the platform, the chip with thousands of isolated microgels holds the capability of self-absorption and partition of DNA samples, thus avoiding auxiliary equipment and professional personnel operations. Using the integrated dLAMP platform, λDNA templates have been quantified with a good linear detection range of 2-1000 copies/μL and a detection limit of 1 copy/μL. As a demonstration, the epidermal growth factor receptor L858R gene mutation, a crucial factor for the susceptibility of the tyrosine kinase inhibitor in non-small-cell lung cancer treatment, has been accurately identified by the dLAMP platform with a spiked plasma sample. This work shows that the developed dLAMP platform provides a low-cost, facile, and user-friendly solution for the absolute quantification of DNA, showing great potential for the POCT of nucleic acids.
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