Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI, United States.
Department of Agricultural Biosystems Engineering, Michigan State University, East Lansing, MI, United States.
Biosens Bioelectron. 2018 Mar 15;101:29-36. doi: 10.1016/j.bios.2017.10.011. Epub 2017 Oct 7.
The interaction between gold nanoparticles (AuNPs) and nucleic acids has facilitated a variety of diagnostic applications, with further diversification of synthesis match bio-applications while reducing biotoxicity. However, DNA interactions with unique surface capping agents have not been fully defined. Using dextrin-capped AuNPs (d-AuNPs), we have developed a novel unamplified genomic DNA (gDNA) nanosensor, exploiting dispersion and aggregation characteristics of d-AuNPs, in the presence of gDNA, for sequence-specific detection. We demonstrate that d-AuNPs are stable in a five-fold greater salt concentration than citrate-capped AuNPs and the d-AuNPs were stabilized by single stranded DNA probe (ssDNAp). However, in the elevated salt concentrations of the DNA detection assay, the target reactions were surprisingly further stabilized by the formation of a ssDNAp-target gDNA complex. The results presented herein lead us to propose a mechanism whereby genomic ssDNA secondary structure formation during ssDNAp-to-target gDNA binding enables d-AuNP stabilization in elevated ionic environments. Using the assay described herein, we were successful in detecting as little as 2.94 fM of pathogen DNA, and using crude extractions of a pathogen matrix, as few as 18 spores/µL.
金纳米粒子 (AuNPs) 与核酸之间的相互作用促进了各种诊断应用,进一步多样化了合成与生物应用的匹配,同时降低了生物毒性。然而,与独特的表面封端剂的 DNA 相互作用尚未完全定义。使用糊精封端的 AuNPs (d-AuNPs),我们开发了一种新型的未扩增基因组 DNA (gDNA) 纳米传感器,利用 d-AuNPs 的分散和聚集特性,在 gDNA 存在的情况下,进行序列特异性检测。我们证明,d-AuNPs 在比柠檬酸封端的 AuNPs 高五倍的盐浓度下稳定,并且 d-AuNPs 由单链 DNA 探针 (ssDNAp) 稳定。然而,在 DNA 检测分析的高盐浓度下,令人惊讶的是,目标反应通过形成 ssDNAp-靶 gDNA 复合物进一步得到稳定。本文提出了一种机制,即在 ssDNAp 与靶 gDNA 结合过程中基因组 ssDNA 二级结构的形成,使 d-AuNP 在高离子环境中得到稳定。使用本文所述的测定方法,我们成功地检测到低至 2.94 fM 的病原体 DNA,并且使用病原体基质的粗提物,低至 18 个孢子/µL。