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Tytuł:
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An integrated electrochemical biosensor based on target-triggered strand displacement amplification and "four-way" DNA junction towards ultrasensitive detection of PIK3CA gene mutation.
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Autorzy:
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Wang T; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Peng Q; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China; Basic Medical College, Chongqing Medical University, Chongqing, 400016, China.
Guo B; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Zhang D; Department of Laboratory Diagnosis, The Third Affiliated Hospital of Shenzhen University, Shenzhen University, Shenzhen, 518000, China.
Zhao M; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Que H; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Wu H; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Yan Y; Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China. Electronic address: .
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Źródło:
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Biosensors & bioelectronics [Biosens Bioelectron] 2020 Feb 15; Vol. 150, pp. 111954. Date of Electronic Publication: 2019 Dec 06.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Publication: Oxford : Elsevier Advanced Technology
Original Publication: [Barking, Essex, England] : Elsevier Applied Science, 1989-
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MeSH Terms:
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Point Mutation*
Class I Phosphatidylinositol 3-Kinases/*genetics
DNA/*genetics
Biosensing Techniques/methods ; Electrochemical Techniques/methods ; Humans ; Nucleic Acid Amplification Techniques/methods
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Contributed Indexing:
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Keywords: Electrochemical biosensor; Four-way DNA junction; Methylene blue; NsbI-SDA; PIK3CA gene mutation
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Substance Nomenclature:
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9007-49-2 (DNA)
EC 2.7.1.137 (Class I Phosphatidylinositol 3-Kinases)
EC 2.7.1.137 (PIK3CA protein, human)
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Entry Date(s):
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Date Created: 20200114 Date Completed: 20201123 Latest Revision: 20201123
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Update Code:
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20240105
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DOI:
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10.1016/j.bios.2019.111954
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PMID:
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31929087
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A novel electrochemical biosensor was constructed for specific and ultrasensitive detection of PIK3CA H1047R gene mutation based on NsbI restriction enzyme-mediated strand displacement amplification (NsbI-SDA) and four-way DNA junction for the first time. In this biosensor, the NsbI restriction enzyme combined with strand displacement amplification (SDA) was able to specifically distinguish PIK3CA H1047R gene mutation and increase the number of DNA copies to improve electrochemical response. In the presence of target mutation gene, DNA fragments produced by the cleavage event of NsbI restriction enzyme could trigger the SDA reaction to generate massive linker chains. When the linker chains were captured on the electrode, the four-way DNA junction was then attached at the end of linker chain. By integrating electroactive molecules of methylene blue (MB) into four-way DNA junction, this sandwich-like electrochemical biosensor was able to determine the specific distinction of target mutation gene with a low detection limit of 0.001%. Finally, this strategy could be used to analyze mutation gene spiked into human serum samples, indicating the potential application in genetic analysis and clinical disease diagnosis.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
