A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex
In this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL) detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease)-assisted target recycling amplification (TRA), rolling circle amplification (RCA) and hemin/G...
| Main Authors: | , , , , , |
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| Format: | Article |
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MDPI AG
2015-01-01
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| Series: | Sensors |
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| Online Access: | http://www.mdpi.com/1424-8220/15/2/2629 |
| _version_ | 1811187736937234432 |
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| author | Fukang Luo Guimin Xiang Xiaoyun Pu Juanchun Yu Ming Chen Guohui Chen |
| author_facet | Fukang Luo Guimin Xiang Xiaoyun Pu Juanchun Yu Ming Chen Guohui Chen |
| author_sort | Fukang Luo |
| collection | DOAJ |
| description | In this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL) detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease)-assisted target recycling amplification (TRA), rolling circle amplification (RCA) and hemin/G-quadruplex. The target DNAs can hybridize with self-assembled capture probes and assistant probes to form “Y” junction structures on the electrode surface, thus triggering the execution of a TRA reaction with the aid of Nt.BbvCI. Then, the RCA reaction and the addition of hemin result in the production of numerous hemin/G-quadruplex, which consume the dissolved oxygen in the detection buffer and result in a significant ECL quenching effect toward the O2/S2O82− system. The proposed strategy combines the amplification ability of TRA, RCA and the inherent high sensitivity of the ECL technique, thus enabling low aM (3.8 aM) detection for sequence-specific DNA and a wide linear range from 10.0 aM to 1.0 pM. At the same time, this novel strategy shows high selectivity against single-base mismatch sequences, which makes our novel universal and highly sensitive method a powerful addition to specific DNA sequence detection. |
| first_indexed | 2024-04-11T14:08:33Z |
| format | Article |
| id | doaj.art-cc19893c51924038900cc18bfcf3e9b7 |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-11T14:08:33Z |
| publishDate | 2015-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Sensors |
| spelling | doaj.art-cc19893c51924038900cc18bfcf3e9b72022-12-22T04:19:49ZengMDPI AGSensors1424-82202015-01-011522629264310.3390/s150202629s150202629A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-QuadruplexFukang Luo0Guimin Xiang1Xiaoyun Pu2Juanchun Yu3Ming Chen4Guohui Chen5Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, ChinaDepartment of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, ChinaDepartment of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, ChinaDepartment of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, ChinaDepartment of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, ChinaDepartment of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, ChinaIn this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL) detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease)-assisted target recycling amplification (TRA), rolling circle amplification (RCA) and hemin/G-quadruplex. The target DNAs can hybridize with self-assembled capture probes and assistant probes to form “Y” junction structures on the electrode surface, thus triggering the execution of a TRA reaction with the aid of Nt.BbvCI. Then, the RCA reaction and the addition of hemin result in the production of numerous hemin/G-quadruplex, which consume the dissolved oxygen in the detection buffer and result in a significant ECL quenching effect toward the O2/S2O82− system. The proposed strategy combines the amplification ability of TRA, RCA and the inherent high sensitivity of the ECL technique, thus enabling low aM (3.8 aM) detection for sequence-specific DNA and a wide linear range from 10.0 aM to 1.0 pM. At the same time, this novel strategy shows high selectivity against single-base mismatch sequences, which makes our novel universal and highly sensitive method a powerful addition to specific DNA sequence detection.http://www.mdpi.com/1424-8220/15/2/2629target recycling amplification (TRA)rolling circle amplification (RCA)hemin/G-quadruplexNt.BbvCIECL |
| spellingShingle | Fukang Luo Guimin Xiang Xiaoyun Pu Juanchun Yu Ming Chen Guohui Chen A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex Sensors target recycling amplification (TRA) rolling circle amplification (RCA) hemin/G-quadruplex Nt.BbvCI ECL |
| title | A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex |
| title_full | A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex |
| title_fullStr | A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex |
| title_full_unstemmed | A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex |
| title_short | A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex |
| title_sort | novel ultrasensitive ecl sensor for dna detection based on nicking endonuclease assisted target recycling amplification rolling circle amplification and hemin g quadruplex |
| topic | target recycling amplification (TRA) rolling circle amplification (RCA) hemin/G-quadruplex Nt.BbvCI ECL |
| url | http://www.mdpi.com/1424-8220/15/2/2629 |
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