Yu Xu, Zhang Zhi-Ling, Zheng Si-Yang
Micro & Nano Integrated Biosystem (MINIBio) Laboratory, Department of Biomedical Engineering, The Pennsylvania State University, University Park, PA 16802, USA.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, and State Key Laboratory of Virology, Wuhan University, Wuhan, Hubei 430072, China.
Biosens Bioelectron. 2015 Apr 15;66:520-6. doi: 10.1016/j.bios.2014.11.035. Epub 2014 Nov 26.
A novel highly sensitive colorimetric assay for DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization was established. The DNA modified superparamagnetic beads were demonstrated to capture and enrich the target DNA in the hybridization buffer or human plasma. The hybridization chain reaction and enzyme-induced silver metallization on the gold nanoparticles were used as cascade signal amplification for the detection of target DNA. The metalization of silver on the gold nanoparticles induced a significant color change from red to yellow until black depending on the concentration of the target DNA, which could be recognized by naked eyes. This method showed a good specificity for the target DNA detection, with the capabilty to discriminate single-base-pair mismatched DNA mutation (single nucleotide polymorphism). Meanwhile, this approach exhibited an excellent anti-interference capability with the convenience of the magentic seperation and washing, which enabled its usage in complex biological systems such as human blood plasma. As an added benefit, the utilization of hybridization chain reaction and enzyme-induced metallization improved detection sensitivity down to 10pM, which is about 100-fold lower than that of traditional unamplified homogeneous assays.
建立了一种基于杂交链式反应和酶诱导金属化的级联扩增策略的新型高灵敏度比色法用于DNA检测。经证明,DNA修饰的超顺磁性微珠可在杂交缓冲液或人血浆中捕获并富集目标DNA。金纳米颗粒上的杂交链式反应和酶诱导的银金属化用作检测目标DNA的级联信号放大。金纳米颗粒上银的金属化会导致显著的颜色变化,从红色变为黄色直至黑色,这取决于目标DNA的浓度,肉眼即可识别。该方法对目标DNA检测具有良好的特异性,能够区分单碱基对错配的DNA突变(单核苷酸多态性)。同时,该方法具有出色的抗干扰能力,并且磁分离和洗涤操作方便,使其能够用于诸如人血浆等复杂生物体系。另外,杂交链式反应和酶诱导金属化的应用将检测灵敏度提高到了10pM,比传统的未扩增均相检测法低约100倍。