Medical College, Henan University of Science and Technology, Luoyang 471003, China; Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
Biosens Bioelectron. 2018 May 15;105:137-142. doi: 10.1016/j.bios.2018.01.034.
By combining the gold nanoparticle (AuNP) enumeration with target-induced strand-displacement reaction, we have developed a non-amplification platform for DNA and miRNA detection based on a deliberately designed sandwich-structured nanocomplex probe (SNC Probe). The proposed strategy can realize the sensitive detection of nucleic acids within 40min with the detection limit of 6.6 fM for HBV DNA and 13.5 fM for miRNA-141, respectively. The method presents reasonable ability to discriminate miRNA-141 from all the other members of miRNA-200 family. And it can also be used for direct detection of miRNA-141 in samples of extracted total small RNA from different cell lines which are reported to have altered levels of miRNA-141. Furthermore, the spike recovery (n = 3) of miRNA-141 in total small RNA extracts of Hela cells is found to be 92.8% for 20 p.M. and 94.7% for 100 p.M. with the standard deviation of 9.2% and 6.8%, respectively. As the only reagent involved in the assay, the SNC Probe presents a very good stability with a relative standard deviation of 3.3% amongst eight tests in 30 days, which greatly simplifies the assay procedure and presents the suitability for routine analyses. On the basis of these findings, this simple non-amplification assay platform can be expected to find assorted applications that can make the best use of the simplicity and sensitivity.
通过将金纳米粒子（AuNP）计数与靶诱导链置换反应相结合，我们开发了一种基于精心设计的三明治结构纳米复合物探针（SNC 探针）的无扩增平台，用于 DNA 和 miRNA 的检测。该策略可以在 40min 内实现核酸的灵敏检测，HBV DNA 的检测限为 6.6fM，miRNA-141 的检测限为 13.5fM。该方法具有合理的能力来区分 miRNA-141 与 miRNA-200 家族的所有其他成员。它还可以用于直接检测不同细胞系中提取的总小 RNA 中的 miRNA-141，这些细胞系被报道具有改变的 miRNA-141 水平。此外，在 HeLa 细胞总小 RNA 提取物中，miRNA-141 的 spike 回收率（n = 3）对于 20pM 为 92.8%，对于 100pM 为 94.7%，标准偏差分别为 9.2%和 6.8%。作为该测定中唯一涉及的试剂，SNC 探针具有非常好的稳定性，在 30 天内进行了 8 次测试，其相对标准偏差为 3.3%，这大大简化了测定程序，并具有常规分析的适用性。基于这些发现，这种简单的无扩增检测平台有望找到各种应用，从而充分利用其简单性和灵敏度。
