In this paper, we designed and evaluated a duplex detection strategy for micro RNAs（mi RNAs） using universal probe-based target-triggered double hybridization and fluorescent microsphere-based assay system（x MAP ar...In this paper, we designed and evaluated a duplex detection strategy for micro RNAs（mi RNAs） using universal probe-based target-triggered double hybridization and fluorescent microsphere-based assay system（x MAP array）. In the absence of target mi RNA, reporter DNA cannot hybridize stably with the immobilized capture DNA due to its low melting temperature. Only after adding target mi RNA, can reporter probe hybridize with capture probe to form a stable three-component complex. This targettriggered stable hybridization makes this method possible for highly selective and sensitive detection of multiple mi RNAs. We exemplified a quantitative detection of duplex mi RNAs with a limit of detection of40 p M. The x MAP array platform holds the potential of extending this approach to simultaneous detection of up to 100 mi RNA targets. Considering the simplicity, rapidity and multiplexing, this work promised a potential detection of multiple mi RNA biomarkers for early disease diagnosis and prognosis.展开更多
Selective and sensitive detection of trace microRNA is important for early diagnosis of diseases due to its expression level related to diseases.Herein,a triple signal amplification strategy is developed for trace mic...Selective and sensitive detection of trace microRNA is important for early diagnosis of diseases due to its expression level related to diseases.Herein,a triple signal amplification strategy is developed for trace microRNA-21 (miRNA-21) detection by combining with target-triggered cyclic strand displacement reaction (TCSDR),hybridization chain reaction (HCR) and enzyme catalytic amplification.Four DNA hairpins(H1,H2,H3,H4) are employed to form an ultralong double-strand DNA (dsDNA) structure,which is initiated by target miRNA-21.As H3 and H4 are labeled with horseradish peroxidase (HRP),numerous HRPs are loaded on the long dsDNA,producing significantly enhanced electrocatalytic signals in the hydrogen peroxide (H_(2)O_(2)) and 3,3,5,5-tetramethylbenzidine (TMB) reaction strategy.Compared with single signal amplification,the triple signal amplification strategy shows higher electrochemical response,wider dynamic range and lower detection limit for miRNA-21 detection with excellent selectivity,reproducibility and stability.Taking advantage of the triple signal amplification strategy,the proposed electrochemical biosensor can detect miRNA-21 in 10 He La cell lysates,suggesting that it is a promising method for fruitful assay in clinical diagnosis.展开更多
基金financially supported by the National Science Foundation of China (Grant No. 21575029)
文摘In this paper, we designed and evaluated a duplex detection strategy for micro RNAs（mi RNAs） using universal probe-based target-triggered double hybridization and fluorescent microsphere-based assay system（x MAP array）. In the absence of target mi RNA, reporter DNA cannot hybridize stably with the immobilized capture DNA due to its low melting temperature. Only after adding target mi RNA, can reporter probe hybridize with capture probe to form a stable three-component complex. This targettriggered stable hybridization makes this method possible for highly selective and sensitive detection of multiple mi RNAs. We exemplified a quantitative detection of duplex mi RNAs with a limit of detection of40 p M. The x MAP array platform holds the potential of extending this approach to simultaneous detection of up to 100 mi RNA targets. Considering the simplicity, rapidity and multiplexing, this work promised a potential detection of multiple mi RNA biomarkers for early disease diagnosis and prognosis.
基金supported by the National Key Research and Development Program of China (No. 2017YFA0205302)the Natural Science Foundation of Jiangsu Province-Major Project (No. BK20212012)+2 种基金the National Natural Science Foundation of China (No. 21874071)the “Six Talents Peak” Foundation of Jiangsu Province (No. SWYY-046)the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD, No. YX030003)。
文摘Selective and sensitive detection of trace microRNA is important for early diagnosis of diseases due to its expression level related to diseases.Herein,a triple signal amplification strategy is developed for trace microRNA-21 (miRNA-21) detection by combining with target-triggered cyclic strand displacement reaction (TCSDR),hybridization chain reaction (HCR) and enzyme catalytic amplification.Four DNA hairpins(H1,H2,H3,H4) are employed to form an ultralong double-strand DNA (dsDNA) structure,which is initiated by target miRNA-21.As H3 and H4 are labeled with horseradish peroxidase (HRP),numerous HRPs are loaded on the long dsDNA,producing significantly enhanced electrocatalytic signals in the hydrogen peroxide (H_(2)O_(2)) and 3,3,5,5-tetramethylbenzidine (TMB) reaction strategy.Compared with single signal amplification,the triple signal amplification strategy shows higher electrochemical response,wider dynamic range and lower detection limit for miRNA-21 detection with excellent selectivity,reproducibility and stability.Taking advantage of the triple signal amplification strategy,the proposed electrochemical biosensor can detect miRNA-21 in 10 He La cell lysates,suggesting that it is a promising method for fruitful assay in clinical diagnosis.
