Attomolar sensitivity microRNA detection using real-time digital microarrays.
Attomolar sensitivity microRNA detection using real-time digital microarrays.
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DOI：
10.1038/s41598-022-19912-z
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发表时间：
2022-09-28
影响因子：
4.6
通讯作者：
Unlu, M. Selim
中科院分区：
文献类型：
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作者：
Kanik, Fulya Ekiz;Celebi, Iris;Sevenler, Derin;Tanriverdi, Kahraman;Unlu, Nese Lortlar;Freedman, Jane E.;Unlu, M. Selim
MicroRNAs （miRNAs）是一类非编码的功能性rna。随着分子生物学的最新发展，miRNA检测引起了极大的兴趣，因为数百种miRNA及其表达水平已被证明与各种疾病（如感染、心血管疾病和癌症）有关。DNA微阵列技术是一种强大的、高通量的核酸检测工具。然而，传统的方法在灵敏度和特异性上不能满足要求，这对miRNA检测在诊断应用中的适应性提出了重大挑战。在这项研究中，我们开发了一个高灵敏度和多路复用的数字微阵列，使用等离子体金纳米棒作为标签。为了验证概念研究，我们使用miRNA-451a （miR-451）和miRNA-223-3p （miR-223）两种mirna进行了实验。通过实现对传感器表面目标分子的实时跟踪，与采用固相支持捕获的传统终点分析相比，我们证明了灵敏度的提高。粒子跟踪克服了在存在低亲和力但高丰度背景分子的情况下检测低丰度生物标志物的灵敏度限制，其中终点分析不足。测得的绝对最低浓度为100 aM。测量的检测限远高于空白样品，我们对外推检测限（LOD）进行了理论计算。动态跟踪将两种mirna的外推lod从飞摩尔范围提高到10原子摩尔（0.2 ml样品中少于1300个拷贝），总孵卵时间从5小时减少到35分钟。
MicroRNAs (miRNAs) are a family of noncoding, functional RNAs. With recent developments in molecular biology, miRNA detection has attracted significant interest, as hundreds of miRNAs and their expression levels have shown to be linked to various diseases such as infections, cardiovascular disorders and cancers. A powerful and high throughput tool for nucleic acid detection is the DNA microarray technology. However, conventional methods do not meet the demands in sensitivity and specificity, presenting significant challenges for the adaptation of miRNA detection for diagnostic applications. In this study, we developed a highly sensitive and multiplexed digital microarray using plasmonic gold nanorods as labels. For proof of concept studies, we conducted experiments with two miRNAs, miRNA-451a (miR-451) and miRNA-223-3p (miR-223). We demonstrated improvements in sensitivity in comparison to traditional end-point assays that employ capture on solid phase support, by implementing real-time tracking of the target molecules on the sensor surface. Particle tracking overcomes the sensitivity limitations for detection of low-abundance biomarkers in the presence of low-affinity but high-abundance background molecules, where endpoint assays fall short. The absolute lowest measured concentration was 100 aM. The measured detection limit being well above the blank samples, we performed theoretical calculations for an extrapolated limit of detection (LOD). The dynamic tracking improved the extrapolated LODs from femtomolar range to 10 attomolar (less than 1300 copies in 0.2 ml of sample) for both miRNAs and the total incubation time was decreased from 5 h to 35 min.
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影响因子：
5.5
作者：
Detassis S;Grasso M;Del Vescovo V;Denti MA
通讯作者：
Denti MA
影响因子：
6.1
作者：
Castell-Auvi, Anna;Cedo, Lidia;Ardevol, Anna
通讯作者：
Ardevol, Anna
影响因子：
5.6
作者：
Gerry, NP;Witowski, NE;Barany, F
通讯作者：
Barany, F
DOI：
10.1073/pnas.1904770116
发表时间：
2019-09-24
影响因子：
11.1
作者：
Canady, Taylor D.;Li, Nantao;Cunningham, Brian T.
通讯作者：
Cunningham, Brian T.
影响因子：
82.9
作者：
通讯作者：
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