Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal

Abstract Alterations in microRNA (miRNA) expression profiles play a pivotal role in the initiation and progression of various diseases, including pediatric pneumonia. Consequently, the development of sensitive, specific, and precise methodologies for miRNA detection represents a promising tool for c...

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Main Authors: Zhe Zhang, Qiaoyi Xie, Chenbo Zhu, Huali Shao, Boying Wu
Format: Article
Language:English
Published: SpringerOpen 2025-08-01
Series:Journal of Analytical Science and Technology
Subjects:
Online Access:https://doi.org/10.1186/s40543-025-00506-z
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author Zhe Zhang
Qiaoyi Xie
Chenbo Zhu
Huali Shao
Boying Wu
author_facet Zhe Zhang
Qiaoyi Xie
Chenbo Zhu
Huali Shao
Boying Wu
author_sort Zhe Zhang
collection DOAJ
description Abstract Alterations in microRNA (miRNA) expression profiles play a pivotal role in the initiation and progression of various diseases, including pediatric pneumonia. Consequently, the development of sensitive, specific, and precise methodologies for miRNA detection represents a promising tool for clinical diagnosis and therapeutic intervention. This study proposes a highly sensitive miRNA detection approach utilizing catalytic hairpin assembly (CHA)-mediated probe release through chain extension and cascade signal amplification. The method enables precise discrimination of target miRNAs from other RNA species via a dual-target recognition mechanism. Furthermore, the incorporation of four signal amplification cycles confers substantial amplification efficiency, permitting accurate detection of low-abundance miRNAs. This system employs cascade isothermal amplification, demonstrating a linear response across concentrations from 1 fM to 100 pM and achieving a detection limit of 0.54 fM. The methodology exhibits significant potential for practical application in complex biological samples, providing a novel platform for profiling miRNA expression patterns and elucidating their mechanistic roles in disease pathogenesis.
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institution Kabale University
issn 2093-3371
language English
publishDate 2025-08-01
publisher SpringerOpen
record_format Article
series Journal of Analytical Science and Technology
spelling doaj-art-6ccbdf9ba7d94b8298be9dcd5bb7054c2025-08-20T04:02:57ZengSpringerOpenJournal of Analytical Science and Technology2093-33712025-08-011611710.1186/s40543-025-00506-zLoading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signalZhe Zhang0Qiaoyi Xie1Chenbo Zhu2Huali Shao3Boying Wu4Department of Pediatrics, Yinzhou District, The Affiliated People’s Hospital of Ningbo UniversityDepartment of Pediatrics, Yinzhou District, The Affiliated People’s Hospital of Ningbo UniversityDepartment of Pediatrics, Yinzhou District, The Affiliated People’s Hospital of Ningbo UniversityDepartment of Pediatrics, Yinzhou District, The Affiliated People’s Hospital of Ningbo UniversityDepartment of Pediatrics, Yinzhou District, The Affiliated People’s Hospital of Ningbo UniversityAbstract Alterations in microRNA (miRNA) expression profiles play a pivotal role in the initiation and progression of various diseases, including pediatric pneumonia. Consequently, the development of sensitive, specific, and precise methodologies for miRNA detection represents a promising tool for clinical diagnosis and therapeutic intervention. This study proposes a highly sensitive miRNA detection approach utilizing catalytic hairpin assembly (CHA)-mediated probe release through chain extension and cascade signal amplification. The method enables precise discrimination of target miRNAs from other RNA species via a dual-target recognition mechanism. Furthermore, the incorporation of four signal amplification cycles confers substantial amplification efficiency, permitting accurate detection of low-abundance miRNAs. This system employs cascade isothermal amplification, demonstrating a linear response across concentrations from 1 fM to 100 pM and achieving a detection limit of 0.54 fM. The methodology exhibits significant potential for practical application in complex biological samples, providing a novel platform for profiling miRNA expression patterns and elucidating their mechanistic roles in disease pathogenesis.https://doi.org/10.1186/s40543-025-00506-zMicroRNACatalytic hairpin assemblyDNA polymeraseEndonucleasePediatric pneumonia
spellingShingle Zhe Zhang
Qiaoyi Xie
Chenbo Zhu
Huali Shao
Boying Wu
Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal
Journal of Analytical Science and Technology
MicroRNA
Catalytic hairpin assembly
DNA polymerase
Endonuclease
Pediatric pneumonia
title Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal
title_full Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal
title_fullStr Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal
title_full_unstemmed Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal
title_short Loading scaffold-mediated multiple signal cycle for ultrasensitive microRNA analysis with low background signal
title_sort loading scaffold mediated multiple signal cycle for ultrasensitive microrna analysis with low background signal
topic MicroRNA
Catalytic hairpin assembly
DNA polymerase
Endonuclease
Pediatric pneumonia
url https://doi.org/10.1186/s40543-025-00506-z
work_keys_str_mv AT zhezhang loadingscaffoldmediatedmultiplesignalcycleforultrasensitivemicrornaanalysiswithlowbackgroundsignal
AT qiaoyixie loadingscaffoldmediatedmultiplesignalcycleforultrasensitivemicrornaanalysiswithlowbackgroundsignal
AT chenbozhu loadingscaffoldmediatedmultiplesignalcycleforultrasensitivemicrornaanalysiswithlowbackgroundsignal
AT hualishao loadingscaffoldmediatedmultiplesignalcycleforultrasensitivemicrornaanalysiswithlowbackgroundsignal
AT boyingwu loadingscaffoldmediatedmultiplesignalcycleforultrasensitivemicrornaanalysiswithlowbackgroundsignal