A ratiometric fluorescent aptamer assay based on RCA amplification mult-G-quadruplex

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Analytical and Bioanalytical Chemistry Pub Date : 2025-08-27 DOI:10.1007/s00216-025-06083-9

Xinyu Xie, Weiling Li, Zhiguang Suo, Rui Guo, Min Wei, Huali Jin

{"title":"A ratiometric fluorescent aptamer assay based on RCA amplification mult-G-quadruplex","authors":"Xinyu Xie, Weiling Li, Zhiguang Suo, Rui Guo, Min Wei, Huali Jin","doi":"10.1007/s00216-025-06083-9","DOIUrl":null,"url":null,"abstract":"<div><p>Ratiometric assay with dual signals can effectively avoid false positives of single signals. Therefore, a dual-signal assay was designed to detect Aflatoxin B1 (AFB1) using the affinity of the aptamers to the target as well as the binding force between the DNA strands. The first fluorescent signal was obtained by binding AFB1 to aptamer to shed the complementary sequence with the FAM (6-carboxyfluorescein). The G-quadruplex is a common DNA conformation capable of emitting light with special fluorescent dyes. At room temperature, the G-quadruplex is opened by the shed complementary strand to form double-stranded DNA, disrupting its original structure. The NMM (N-methylmesoporphyrin IX) dye is unable to embed the G-quadruplex and emit light. Taking advantage of the DNA conformational transition, NMM dye was introduced as the second fluorescence signal. Based on the above principle, a dual fluorescence signal ratiometric assay was constructed for the detection of AFB1 by the ratio of FAM and NMM fluorescence intensity. This study provides a prospective strategy for developing a dual-signal construct ratio assay.</p></div>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":"417 26","pages":"5885 - 5893"},"PeriodicalIF":3.8000,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical and Bioanalytical Chemistry","FirstCategoryId":"92","ListUrlMain":"https://link.springer.com/article/10.1007/s00216-025-06083-9","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 0

Abstract

Ratiometric assay with dual signals can effectively avoid false positives of single signals. Therefore, a dual-signal assay was designed to detect Aflatoxin B1 (AFB1) using the affinity of the aptamers to the target as well as the binding force between the DNA strands. The first fluorescent signal was obtained by binding AFB1 to aptamer to shed the complementary sequence with the FAM (6-carboxyfluorescein). The G-quadruplex is a common DNA conformation capable of emitting light with special fluorescent dyes. At room temperature, the G-quadruplex is opened by the shed complementary strand to form double-stranded DNA, disrupting its original structure. The NMM (N-methylmesoporphyrin IX) dye is unable to embed the G-quadruplex and emit light. Taking advantage of the DNA conformational transition, NMM dye was introduced as the second fluorescence signal. Based on the above principle, a dual fluorescence signal ratiometric assay was constructed for the detection of AFB1 by the ratio of FAM and NMM fluorescence intensity. This study provides a prospective strategy for developing a dual-signal construct ratio assay.

查看原文本刊更多论文

基于RCA扩增多g -四重体的比例荧光适体测定。

双信号比例分析法可有效避免单信号的误报。因此，设计了一种双信号检测方法，利用适体对靶标的亲和力以及DNA链之间的结合力检测黄曲霉毒素B1 （AFB1）。第一个荧光信号是通过将AFB1与适体结合，与FAM（6-羧基荧光素）脱落互补序列获得的。g -四重体是一种常见的DNA构象，能够用特殊的荧光染料发光。在室温下，g -四重体被脱落的互补链打开，形成双链DNA，破坏其原有结构。NMM （n -甲基间卟啉IX）染料不能嵌入g -四联体并发光。利用DNA的构象转变，引入NMM染料作为第二荧光信号。基于上述原理，构建FAM与NMM荧光强度之比检测AFB1的双荧光信号比率法。本研究为发展双信号构建比分析提供了一种前瞻性策略。

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

Analytical and Bioanalytical Chemistry 化学-分析化学

CiteScore

8.00

自引率

4.70%

发文量

638

审稿时长

2.1 months

期刊介绍： Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.