Two-stage Nicking Enzyme Signal Amplification Combined with DNAzyme Amplification for the Detection of Bone Morphogenetic Protein 6 mRNA
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- HU Zhaohui
- Department of Orthopaedics, Liuzhou People’s Hospital
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- ZHOU Yanhong
- Department of Orthopaedics, Liuzhou People’s Hospital
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- XIE Xiangtao
- Department of Orthopaedics, Liuzhou People’s Hospital
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- JIANG Renjie
- Department of Orthopaedics, Liuzhou People’s Hospital
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- LI Ningning
- Department of Orthopaedics, Liuzhou People’s Hospital
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Abstract
Highly sensitive detection of bone morphogenetic protein 6 (BMP6) mRNA is essential to monitor bone regeneration in the regenerating defects. In this work, we proposed a quantitative approach based on two-stage nicking enzyme signal amplification (NESA) and DNAzyme amplification for highly sensitive detection of BMP6 mRNA. The two-stage NESA involves two templates and two-stage amplification reactions under isothermal conditions. The first template contains two repeat sequences that could hybridize to the target RNA, triggering an exponential amplification. The amplified product was a short single-stranded DNA with the same sequence as the target RNA. The single-stranded DNA can trigger another linear NESA and produces a large amount of horseradish peroxidase (HRP)-mimicking G-quadruplex DNAzyme. This proposed assay showed a quantitative analysis of BMP6 mRNA in a wide range from 1 fM to 100 nM with a detection limit of 0.01 fM.
Journal
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- Analytical Sciences
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Analytical Sciences 30 (11), 1039-1044, 2014
The Japan Society for Analytical Chemistry
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Details 詳細情報について
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- CRID
- 1390282679236941312
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- NII Article ID
- 130004704335
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- NII Book ID
- AA10500785
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- ISSN
- 13482246
- 09106340
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- NDL BIB ID
- 025902547
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- PubMed
- 25382038
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- Web Site
- https://ndlsearch.ndl.go.jp/books/R000000004-I025902547
- https://link.springer.com/content/pdf/10.2116/analsci.30.1039.pdf
- https://link.springer.com/article/10.2116/analsci.30.1039/fulltext.html
- https://www.jstage.jst.go.jp/article/analsci/30/11/30_1039/_pdf
- https://search.jamas.or.jp/link/ui/2015136891
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed