Digital-resolution and highly sensitive detection of multiple exosomal small RNAs by DNA toehold probe-based photonic resonator absorption microscopy.
Digital-resolution and highly sensitive detection of multiple exosomal small RNAs by DNA toehold probe-based photonic resonator absorption microscopy.
复制标题
DOI：
10.1016/j.talanta.2022.123256
复制
发表时间：
2022-05-01
期刊：
影响因子：
6.1
通讯作者：
Cunningham BT
中科院分区：
文献类型：
--
作者：
Zhao B;Wang W;Li N;Garcia-Lezana T;Che C;Wang X;Losic B;Villanueva A;Cunningham BT
小的非编码RNA（snRNA）已成为检测早期癌症的有前途的诊断生物标志物。目前现有的snRNA检测方法，包括北方印迹、逆转录-聚合酶链反应、微阵列和RNA-Seq，局限于耗时、低灵敏度、昂贵的仪器或复杂的数据分析。在此，我们提出了一种快速定量分析的多个肝癌相关的外泌体snRNA的核酸立足点探针为基础的光子共振吸收显微镜（PRAM）测定，具有数字分辨率和高灵敏度。该测定依赖于使用三个立足点探针编码的金纳米颗粒（AuNP）和可寻址的光子晶体（PC）传感芯片。靶snRNA的存在将引发支点介导的链置换反应，该反应触发金颗粒捕获到PC表面上，随后通过PRAM成像，用于对检测到的snRNA分子进行数字计数。我们实现了高灵敏度和选择性的检测缓冲液中的三个snRNA靶与30分钟的测定方案，与4.56 fM，4.68 fM和0.69 pM的检测限。在确认了我们的测定用于检测掺入到外泌体RNA提取物中的snRNA靶标的性能之后，我们证明了其用于定量检测来自患者血浆样品的相同靶标的能力。该方法提供了一种快速、简单的工作流程，在室温下操作，只需一步，无需酶促扩增，而检测仪器可以作为低成本的便携式系统用于护理点环境。
Small noncoding RNAs (snRNA) have been emerging as promising diagnostic biomarkers for detecting early stage cancer. Currently existing methods for snRNA detection, including northern blot, reverse transcription-polymerase chain reaction, microarrays and RNA-Seq, are limited to time-consuming, low sensitivity, expensive instrumentation or complex analysis of data. Herein, we present a rapid quantitative analysis of multiple liver cancer-associated exosomal snRNA by a nucleic acid toehold probe-based photonic resonator absorption microscopy (PRAM) assay, with digital resolution and high sensitivity. The assay relies on the use of three toehold probe-encoded gold nanoparticles (AuNPs) and addressable photonic crystal (PC) sensing chips. The presence of target snRNA will initiate toehold-mediated strand displacement reactions that trigger the capture of gold particles onto the PC surface, which is subsequently imaged by PRAM for digital counting of detected snRNA molecules. We achieved highly sensitive and selective detection of three snRNA targets in buffer with a 30 minute assay protocol, with detection limits of 4.56 fM, 4.68 fM and 0.69 pM. Having confirmed our assay’s performance for detection of snRNA targets spiked into exosomal RNA extracts, we demonstrated its capability for quantitative detection of the same targets from patient blood plasma samples. The approach offers a rapid, simple workflow that operates at room temperature with a single step without enzymatic amplification, while the detection instrument can be implemented as a low-cost portable system for point of care environments.
登录
查看更多内容
DOI：
10.1007/978-1-4939-7614-0_21
发表时间：
2018-01-01
期刊：
INVESTIGATIONS OF EARLY NUTRITION EFFECTS ON LONG-TERM HEALTH: METHODS AND APPLICATIONS
影响因子：
--
作者：
Pantaleao, Lucas Carminatti;Ozanne, Susan E.
通讯作者：
Ozanne, Susan E.
DOI：
10.1007/978-1-60761-646-7_10
发表时间：
2010-01-01
期刊：
PLANT EPIGENETICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Varkonyi-Gasic, Erika;Hellens, Roger P.
通讯作者：
Hellens, Roger P.
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：
50.3
作者：
Becker A;Thakur BK;Weiss JM;Kim HS;Peinado H;Lyden D
通讯作者：
Lyden D
影响因子：
6.1
作者：
Zhao B;Che C;Wang W;Li N;Cunningham BT
通讯作者：
Cunningham BT