Dong Tao, Yin Ran, Yu Qingcai, Qiu Wanwei, Li Kun, Qian Lisheng, Li He, Shen Bing, Liu Guodong
Institute of Biomedical and Health Science, School of Life and Health Science, Anhui Science and Technology University, Fengyang, Anhui, 233100, China; School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui, 230032, China.
Institute of Biomedical and Health Science, School of Life and Health Science, Anhui Science and Technology University, Fengyang, Anhui, 233100, China; Department of Emergency, Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 533251, China.
Anal Chim Acta. 2021 Feb 22;1147:56-63. doi: 10.1016/j.aca.2020.12.042. Epub 2020 Dec 29.
We report a highly sensitive approach for detecting microRNA-21 (miR-21) in cancer cells and human serum by using Au@Si nanocomposite labeled lateral flow assay. The Au@Si nanocomposite was prepared by coating numerous 3-5 nm gold nanoparticles (GNP) on a silica nanoparticle (SiNP) with a diameter of 150 nm and used as colored label on the lateral flow assay for signal amplification. TEM results show there are around 1000 GNPs coated on the SiNP surface. The principle of miR-21 detection is based on on-strip DNA-microRNA hybridization reactions to form DNA-miR-21-DNA-Au@Si complexes, which are captured on the test zone of the lateral flow test strip and produce a visible red band. A thiol-modified detecting DNA probe (Det-DNA) and a biotin-modified capturing DNA probe (Cap-DNA), which are complementary to miR-21, were used to prepare the lateral flow test strips. After systematic optimization, the method can detect a minimum concentration of 1.0 pM miR-21, which is 60 times lower than that of the GNP-based lateral flow assay (Gao et al. Biosens & Bioelectro, 2014, 54, 578-584). The method was applied to detect miR-21 in cancer cells and spiked human serum with satisfactory results.
我们报道了一种通过使用金@硅纳米复合材料标记的侧向流动分析法来检测癌细胞和人血清中微小RNA - 21（miR - 21）的高灵敏度方法。金@硅纳米复合材料是通过在直径为150纳米的二氧化硅纳米颗粒（SiNP）上包覆大量3 - 5纳米的金纳米颗粒（GNP）制备而成，并用作侧向流动分析法中的显色标记以进行信号放大。透射电子显微镜结果显示，SiNP表面包覆了约1000个GNP。miR - 21的检测原理基于条带上的DNA - 微小RNA杂交反应，形成DNA - miR - 21 - DNA - 金@硅复合物，这些复合物在侧向流动测试条的测试区被捕获并产生可见的红色条带。与miR - 21互补的硫醇修饰检测DNA探针（Det - DNA）和生物素修饰捕获DNA探针（Cap - DNA）被用于制备侧向流动测试条。经过系统优化，该方法可检测到的miR - 21最低浓度为1.0 pM，比基于GNP的侧向流动分析法低60倍（Gao等人，《生物传感器与生物电子学》，2014年，第54卷，第578 - 584页）。该方法应用于检测癌细胞和加标的人血清中的miR - 21，结果令人满意。
