Rolling Circle Amplification as a Molecular Tool for Spatially Resolved Signal Amplification in Single Molecule Counting Assay.

IF 5.6 3区工程技术 Q1 CHEMISTRY, ANALYTICAL

Biosensors-Basel Pub Date : 2025-09-21 DOI:10.3390/bios15090628

Juhwan Park

{"title":"Rolling Circle Amplification as a Molecular Tool for Spatially Resolved Signal Amplification in Single Molecule Counting Assay.","authors":"Juhwan Park","doi":"10.3390/bios15090628","DOIUrl":null,"url":null,"abstract":"<p><p>There have been rising interests in ultra-sensitive biosensing technologies for early diagnosis and prognosis monitoring of infectious diseases, cancers, and neurodegenerative diseases. Digital signal readout strategy represented by digital ELISA or digital PCR, advanced biosensing field enormously, which enables detection of biomolecules under the detection limit of conventional biosensing methods. However, due to the need for compartmentalization and limited multiplex capability, it has been hurdled for utilization in applications requiring hierarchical resolution analysis such as sub-cellular molecules or molecular cargo of single cells or single extracellular vesicles (EVs). Rolling circle amplification (RCA), an isothermal DNA amplification method enabling localization of an amplified signal, can eliminate the need for compartmentalization and increase multiplex capability. It also has potential to expand applications of single molecule counting assay for understanding hierarchy of biological systems. In this review, recent advances in RCA-based single molecule counting assay are overviewed and their applications in single cells and single EVs quantitative analysis are discussed. Furthermore, the limitations and outlook of RCA-based single molecule counting assay are highlighted.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":"15 9","pages":""},"PeriodicalIF":5.6000,"publicationDate":"2025-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12468232/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biosensors-Basel","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.3390/bios15090628","RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}

引用次数: 0

Abstract

There have been rising interests in ultra-sensitive biosensing technologies for early diagnosis and prognosis monitoring of infectious diseases, cancers, and neurodegenerative diseases. Digital signal readout strategy represented by digital ELISA or digital PCR, advanced biosensing field enormously, which enables detection of biomolecules under the detection limit of conventional biosensing methods. However, due to the need for compartmentalization and limited multiplex capability, it has been hurdled for utilization in applications requiring hierarchical resolution analysis such as sub-cellular molecules or molecular cargo of single cells or single extracellular vesicles (EVs). Rolling circle amplification (RCA), an isothermal DNA amplification method enabling localization of an amplified signal, can eliminate the need for compartmentalization and increase multiplex capability. It also has potential to expand applications of single molecule counting assay for understanding hierarchy of biological systems. In this review, recent advances in RCA-based single molecule counting assay are overviewed and their applications in single cells and single EVs quantitative analysis are discussed. Furthermore, the limitations and outlook of RCA-based single molecule counting assay are highlighted.

Abstract Image

查看原文本刊更多论文

滚动圆放大作为单分子计数分析中空间分辨信号放大的分子工具。

人们对用于传染病、癌症和神经退行性疾病的早期诊断和预后监测的超灵敏生物传感技术越来越感兴趣。以数字ELISA或数字PCR为代表的数字信号读出策略，极大地推动了生物传感领域的发展，可以在传统生物传感方法的检测极限下检测生物分子。然而，由于需要分隔和有限的多重功能，它在需要分层分辨率分析的应用中受到阻碍，例如亚细胞分子或单个细胞或单个细胞外囊泡（ev）的分子货物。滚动圈扩增（RCA）是一种能够定位扩增信号的等温DNA扩增方法，它可以消除对区隔化的需要并提高多路传输能力。它也有可能扩大单分子计数分析在了解生物系统层次结构方面的应用。本文综述了基于rca的单分子计数技术的最新进展，并对其在单细胞和单ev定量分析中的应用进行了讨论。并对基于rca的单分子计数方法的局限性和前景进行了展望。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Biosensors-Basel Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry

CiteScore

6.60

自引率

14.80%

发文量

983

审稿时长

11 weeks

期刊介绍： Biosensors (ISSN 2079-6374) provides an advanced forum for studies related to the science and technology of biosensors and biosensing. It publishes original research papers, comprehensive reviews and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.