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Cross-Priming-Linked Hierarchical Isothermal Amplification Programming Progressive Activating Clustered Regularly Interspaced Short Palindromic Repeats/Cas12a in miRNA Signaling.
Wang, Qi; Xia, Juan; Yang, Chengying; Chen, Xiumei; Chen, Baoqiang; Li, Yang; Huang, Hong; Lin, Bingyong; Guo, Longhua; Xu, Jianguo.
Afiliación
  • Wang Q; Key Laboratory of Embryo Development and Reproductive Regulation, School of Biological and Food Engineering, Fuyang Normal University, Fuyang, Anhui 236037, P. R. China.
  • Xia J; Jiaxing Key Laboratory of Molecular Recognition and Sensing, College of Biological, Chemical Sciences and Engineering, Jiaxing University, Jiaxing, Zhejiang 314001, P. R. China.
  • Yang C; Key Laboratory of Embryo Development and Reproductive Regulation, School of Biological and Food Engineering, Fuyang Normal University, Fuyang, Anhui 236037, P. R. China.
  • Chen X; Key Laboratory of Embryo Development and Reproductive Regulation, School of Biological and Food Engineering, Fuyang Normal University, Fuyang, Anhui 236037, P. R. China.
  • Chen B; Key Laboratory of Embryo Development and Reproductive Regulation, School of Biological and Food Engineering, Fuyang Normal University, Fuyang, Anhui 236037, P. R. China.
  • Li Y; Jiaxing Key Laboratory of Molecular Recognition and Sensing, College of Biological, Chemical Sciences and Engineering, Jiaxing University, Jiaxing, Zhejiang 314001, P. R. China.
  • Huang H; Key Laboratory of Embryo Development and Reproductive Regulation, School of Biological and Food Engineering, Fuyang Normal University, Fuyang, Anhui 236037, P. R. China.
  • Lin B; Jiaxing Key Laboratory of Molecular Recognition and Sensing, College of Biological, Chemical Sciences and Engineering, Jiaxing University, Jiaxing, Zhejiang 314001, P. R. China.
  • Guo L; Jiaxing Key Laboratory of Molecular Recognition and Sensing, College of Biological, Chemical Sciences and Engineering, Jiaxing University, Jiaxing, Zhejiang 314001, P. R. China.
  • Xu J; Jiaxing Key Laboratory of Molecular Recognition and Sensing, College of Biological, Chemical Sciences and Engineering, Jiaxing University, Jiaxing, Zhejiang 314001, P. R. China.
Anal Chem ; 96(35): 14205-14214, 2024 Sep 03.
Article en En | MEDLINE | ID: mdl-39171996
ABSTRACT
Cascade isothermal nucleic acid amplification, which integrates several different amplification protocols to enhance the assay performance, is widely utilized in biosensing, particularly for detecting microRNAs (miRNAs), crucial biomarkers associated with tumor initiation and progression. However, striking a balance between a high amplification efficiency and simplicity in design remains a challenge. Therefore, methods achieving high amplification efficiency without significantly increasing complexity are highly favored. In this study, we propose a novel approach for miRNA detection, employing cross-priming-linked hierarchical isothermal amplification (CP-HIA) to progressively activate the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system. The CP-HIA method strategically combines nicking-rolling circle amplification (n-RCA) and palindrome-aided circular strand displacement amplification (p-CSDA) for miRNA detection. Remarkably, this method utilizes only two main probes. Its key innovation lies in the interactive cross-priming strategy, wherein the amplification product from n-RCA is recycled to further drive p-CSDA, and vice versa. This interactive process establishes a hierarchical amplification, significantly enriching the activation probes for progressive CRISPR/Cas12a activation and subsequent target signal amplification. Consequently, the method exhibits greatly enhanced analytical performance, including high sensitivity and specificity in detecting low concentrations of miRNA. As low as 1.06 fM miRNA can thus be quantitatively detected, and the linear response of the miRNA is from 10 fM to 10 nM. These features demonstrate its potential for early disease diagnosis and monitoring. We anticipate that the CP-HIA method will serve as a promising platform for developing advanced molecular diagnostic tools for biomedical research.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Amplificación de Ácido Nucleico / MicroARNs Límite: Humans Idioma: En Revista: Anal Chem Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Amplificación de Ácido Nucleico / MicroARNs Límite: Humans Idioma: En Revista: Anal Chem Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos