Radfar Sasan, Ghanbari Reza, Attaripour Isfahani Ali, Rezaei Halimeh, Kheirollahi Majid
Stem Cell and Regenerative Medicine Center of Excellence, Tehran University of Medical Science, Tehran, Iran.
Department of Biological Science and Technology, Najafabad Branch, Islamic Azad University, Najafabad, Iran.
Bioelectrochemistry. 2022 Jun;145:108087. doi: 10.1016/j.bioelechem.2022.108087. Epub 2022 Feb 17.
Determination of microRNAs (miRNAs) as valuable blood-borne biomarkers has attracted many scientific attentions. However, analytical methods are still restricted by miRNAs intrinsic characteristics. In this study, for the first time, novel blackberry-like magnetic DNA/FMMA nanospheres were synthesized and mounted on a gold stir-bar as signal amplification probes. To produce this strong electrochemical signal label, double strand DNAs were immobilized on gold coated magnetic nanospheres through a hybridization chain reaction followed by reversible addition-fragmentation chain-transfer polymerization, which brought a great quantity of the electroactive tags (FMMA) on the nanosphere surface. These nanospheres were then fixed on the gold stir-bar as signal probes. The magnetic DNA/FMMA nanosphere probes can be released by substituting with the newly emerging DNA fragments of catalyzed hairpin assembly products. Eventually, these signal probes were magnetically enriched on the electrode surface to produce electrochemical signal and finally, the biosensor was developed to detect miRNA-106a (model target). The suggested aptamer-based biosensor demonstrated considerable selectivity, acceptable storage stability, high specificity, and excellent performance in real sample analysis without any pretreatments. As a result, current study reveals that the developed strategy has a great potential for the early diagnosis of gastric cancer and additionally the clinical monitoring of any miRNA sequences.
将微小RNA(miRNA)确定为有价值的血液生物标志物已引起众多科学关注。然而,分析方法仍受miRNA固有特性的限制。在本研究中,首次合成了新型黑莓状磁性DNA/FMMA纳米球,并将其作为信号放大探针固定在金搅拌棒上。为产生这种强电化学信号标记,通过杂交链式反应将双链DNA固定在金包覆的磁性纳米球上,随后进行可逆加成-断裂链转移聚合,这在纳米球表面带来了大量的电活性标签(FMMA)。然后将这些纳米球作为信号探针固定在金搅拌棒上。磁性DNA/FMMA纳米球探针可通过被催化发夹组装产物新出现的DNA片段取代而释放。最终,这些信号探针在电极表面被磁性富集以产生电化学信号,最终开发出用于检测miRNA-106a(模型靶标) 的生物传感器。所建议的基于适配体的生物传感器在无需任何预处理的实际样品分析中表现出相当高的选择性、可接受的储存稳定性、高特异性和优异性能。因此,当前研究表明所开发的策略在胃癌早期诊断以及任何miRNA序列的临床监测方面具有巨大潜力。