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Dual approach for the colorimetric determination of unamplified microRNAs by using citrate capped gold nanoparticles
Microchimica Acta ( IF 5.7 ) Pub Date : 2018-03-22 , DOI: 10.1007/s00604-018-2767-9
Ahmed Ibrahim Nossier , Hana Abdelzaher , Marwa Matboli , Sanaa Eissa

AbstractThe authors describe a method for the colorimetric determination of unamplified microRNA. It is based on the use of citrate-capped gold nanoparticles (AuNPs) and, alternatively, a microRNA-probe hybrid or a magnetically extracted microRNA that serve as stabilizers against the salt-induced aggregation of AuNPs. The absorbance ratios A525/A625 of the reacted AuNP solutions were used to quantify the amount of microRNA. The assay works in the range of 5–25 pmol microRNA. The lower limit of detection (LOD) is 10 pmol. The performance of the method was tested by detection of microRNA-210-3p in totally extracted urinary microRNA from normal, benign, and bladder cancer subjects. The sensitivity and specificity for qualitative detection of urinary microRNA-210-3p using the assay are 74% and 88% respectively, which is consistent with real time PCR based assays. The assay was applied to the determination of specific microRNA by using its specific oligo targeter or following magnetic isolation of the desired microRNA. The method is simple, cost-efficient, has a short turn-around time and requires minimal equipment and personnel. Graphical abstractSchematic of the two detection schemes: In the first approach, matched microRNA hybridizes with its specific probe to stabilize gold nanoparticles (AuNPs) against salt induced aggregation and to leave the red color of the AuNPs unchanged. In the second one, microRNA extracted via magnetic nanoparticles (MNP) stabilizes AuNPs against aggregation.

中文翻译:

使用柠檬酸盐封端的金纳米粒子比色测定未扩增 microRNA 的双重方法

摘要作者描述了一种用于比色测定未扩增 microRNA 的方法。它基于使用柠檬酸盐封端的金纳米粒子 (AuNPs),或者,microRNA 探针杂交或磁性提取的 microRNA,用作稳定剂,防止盐诱导的 AuNPs 聚集。反应后的 AuNP 溶液的吸光度比 A525/A625 用于量化 microRNA 的量。该测定在 5–25 pmol microRNA 的范围内起作用。检测下限 (LOD) 为 10 pmol。该方法的性能通过检测从正常、良性和膀胱癌受试者中完全提取的尿 microRNA 中的 microRNA-210-3p 来测试。使用该测定法定性检测尿液 microRNA-210-3p 的灵敏度和特异性分别为 74% 和 88%,这与基于实时 PCR 的测定一致。通过使用特定的寡核苷酸靶向物或对所需的 microRNA 进行磁性分离,将该测定应用于确定特定的 microRNA。该方法简单、经济、周转时间短,所需设备和人员最少。两种检测方案的图形摘要示意图:在第一种方法中,匹配的 microRNA 与其特异性探针杂交,以稳定金纳米粒子 (AuNPs) 免受盐诱导的聚集,并使 AuNPs 的红色保持不变。在第二个中,通过磁性纳米粒子 (MNP) 提取的 microRNA 可稳定 AuNPs 防止聚集。该方法简单、经济、周转时间短,所需设备和人员最少。两种检测方案的图形摘要示意图:在第一种方法中,匹配的 microRNA 与其特异性探针杂交,以稳定金纳米粒子 (AuNPs) 免受盐诱导的聚集,并使 AuNPs 的红色保持不变。在第二个中,通过磁性纳米粒子 (MNP) 提取的 microRNA 可稳定 AuNPs 防止聚集。该方法简单、经济、周转时间短,所需设备和人员最少。两种检测方案的图形摘要示意图:在第一种方法中,匹配的 microRNA 与其特异性探针杂交,以稳定金纳米粒子 (AuNPs) 免受盐诱导的聚集,并使 AuNPs 的红色保持不变。在第二个中,通过磁性纳米粒子 (MNP) 提取的 microRNA 可稳定 AuNPs 防止聚集。
更新日期:2018-03-22
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