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Stoichiometric Tuning of PNA Probes to Au0.8Ag0.2 Alloy Nanoparticles for Visual Detection of Nucleic Acids in Plasma.
ACS Sensors ( IF 8.2 ) Pub Date : 2020-07-23 , DOI: 10.1021/acssensors.0c00667
Garima Goyal 1, 2, 3 , Gopal Ammanath 2, 3 , Alagappan Palaniappan 2, 3 , Bo Liedberg 1, 2, 3
Affiliation  

Standard detection methods for nucleic acids, an important class of diagnostic biomarkers, are often laborious and cumbersome. In need for development of facile methodologies, localized surface plasmon resonance (LSPR) assays have been widely explored for both spectroscopic and visual detection of nucleic acids. Our sensing approach is based on monitoring changes in the LSPR band due to interaction between peptide nucleic acid (PNA) and plasmonic nanoparticles (NPs) in the presence/absence of target nucleic acid. We have investigated the importance of tuning the stoichiometry of PNA to NPs to enable “naked-eye” detection of nucleic acids at clinically relevant concentration ranges. Assaying in plasma is achieved by incorporation of silver in gold NPs (AuNPs) via an alloying process. The synthesized gold/silver alloy NPs reduce nonspecific adsorption of proteinaceous interferents in plasma. Furthermore, the gold/silver alloy NPs absorb in the most sensitive cyan to green transition zone (∼500 nm) yielding highly competitive visual limits of detection (LODs). The visual LOD (calculated objectively using the ΔE algorithm) for a model microRNA (mir21) using a productive combination of stoichiometric tuning of the PNA to NP ratio and compositional tuning of the NPs in buffer and plasma extract equals 200 pM (∼250 times lower than existing reports) and 3 nM, respectively. We envision that the proposed LSPR assay based on Au0.8Ag0.2NPs offers an avenue for rapid and sensitive on-site detection of nucleic acids in complex matrixes in combination with efficient target extraction kits.

中文翻译:

PNA探针对Au0.8Ag0.2合金纳米粒子的化学计量比调节,用于视觉检测血浆中的核酸。

核酸的标准检测方法是诊断生物标志物的重要类别,通常很费力且麻烦。为了开发简便的方法,已经广泛探索了局部表面等离振子共振(LSPR)测定法用于核酸的光谱学和视觉检测。我们的传感方法基于在目标核酸存在/不存在的情况下,由于多肽核酸(PNA)与等离子体纳米颗粒(NP)之间相互作用而引起的LSPR带变化的监测。我们已经研究了将PNA的化学计量调整为NP的重要性,以使“裸眼”检测在临床相关浓度范围内的核酸成为可能。血浆中的分析是通过合金化工艺将银掺入金NP(AuNP)中来实现的。合成的金/银合金纳米粒子减少了血浆中蛋白质干扰物的非特异性吸附。此外,金/银合金纳米颗粒在最敏感的青色至绿色过渡区(约500 nm)中吸收,从而产生了极具竞争性的视觉检测极限(LOD)。视觉LOD(使用Δ客观计算E算法)用于模型microRNA(mir21),其有效组合是将PNA与NP的化学计量比调整与缓冲液和血浆提取物中NP的成分调整等于200 pM(比现有报告低约250倍)和3 nM,分别。我们设想,基于Au 0.8 Ag 0.2 NPs的LSPR分析方法与有效的目标提取试剂盒相结合,可为复杂基质中核酸的快速,灵敏的现场检测提供一条途径。
更新日期:2020-08-28
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