A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification
A novel homogeneous label-free electrochemical biosensor using G-triplex/methylene blue (G3/MB) complex as the signal generator together with an amplification assisted by the λ-exonuclease (λ-Exo) has been successfully constructed for ultrasensitive microRNA (miRNA) detection. An integrated microele...
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Frontiers Media S.A.
2021
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oai:doaj.org-article:feeccbd594c84535af99635f1f2159a72021-11-04T09:22:27ZA Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification2296-264610.3389/fchem.2021.753253https://doaj.org/article/feeccbd594c84535af99635f1f2159a72021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fchem.2021.753253/fullhttps://doaj.org/toc/2296-2646A novel homogeneous label-free electrochemical biosensor using G-triplex/methylene blue (G3/MB) complex as the signal generator together with an amplification assisted by the λ-exonuclease (λ-Exo) has been successfully constructed for ultrasensitive microRNA (miRNA) detection. An integrated microelectrode was designed to realize the miniaturization of the homogeneous electrochemical assay. Taking advantage of G3, that can specifically bind with MB and decrease its diffusion current, a single-stranded functional DNA hairpin structure was designed as the bio-recognition probe. The probe consisted of G3, eight bases to block G3, and the complementary sequences of the target miRNA. Here we chose miRNA141—a potentially diagnostic biomarker of prostate cancer as the model target. The presence of miRNA141 could hybridize with the probe DNA to form a double-stranded structure with a 5′-phosphorylated terminus. Then λ-Exo was adopted to digest mononucleotides from the 5′-end, leading to the release of G3 part and miRNA141. The released miRNA could hybridize with another probe to trigger the cycling process, while the released G3 could therefore interact with MB to cause a detectable decrease of diffusion current. The proposed strategy showed a low detection limit of 16 fM and an excellent specificity to discriminate single-base mismatches. Furthermore, this sensor was applied to detect miRNA141 from diluted human serum samples, indicating that it has great potential in the application of nucleic acid detection in real samples.Yao MengFangming ChenMingrui JiangQin GuoQin GuoYaqiong WangYaqiong WangJian WangJian WangDe-Wen ZhangDe-Wen ZhangFrontiers Media S.A.articlehomogeneous label-freemicroRNAfunctional nucleic acidsignal amplificationelectrochemical biosensorChemistryQD1-999ENFrontiers in Chemistry, Vol 9 (2021) |
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| collection |
DOAJ |
| language |
EN |
| topic |
homogeneous label-free microRNA functional nucleic acid signal amplification electrochemical biosensor Chemistry QD1-999 |
| spellingShingle |
homogeneous label-free microRNA functional nucleic acid signal amplification electrochemical biosensor Chemistry QD1-999 Yao Meng Fangming Chen Mingrui Jiang Qin Guo Qin Guo Yaqiong Wang Yaqiong Wang Jian Wang Jian Wang De-Wen Zhang De-Wen Zhang A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification |
| description |
A novel homogeneous label-free electrochemical biosensor using G-triplex/methylene blue (G3/MB) complex as the signal generator together with an amplification assisted by the λ-exonuclease (λ-Exo) has been successfully constructed for ultrasensitive microRNA (miRNA) detection. An integrated microelectrode was designed to realize the miniaturization of the homogeneous electrochemical assay. Taking advantage of G3, that can specifically bind with MB and decrease its diffusion current, a single-stranded functional DNA hairpin structure was designed as the bio-recognition probe. The probe consisted of G3, eight bases to block G3, and the complementary sequences of the target miRNA. Here we chose miRNA141—a potentially diagnostic biomarker of prostate cancer as the model target. The presence of miRNA141 could hybridize with the probe DNA to form a double-stranded structure with a 5′-phosphorylated terminus. Then λ-Exo was adopted to digest mononucleotides from the 5′-end, leading to the release of G3 part and miRNA141. The released miRNA could hybridize with another probe to trigger the cycling process, while the released G3 could therefore interact with MB to cause a detectable decrease of diffusion current. The proposed strategy showed a low detection limit of 16 fM and an excellent specificity to discriminate single-base mismatches. Furthermore, this sensor was applied to detect miRNA141 from diluted human serum samples, indicating that it has great potential in the application of nucleic acid detection in real samples. |
| format |
article |
| author |
Yao Meng Fangming Chen Mingrui Jiang Qin Guo Qin Guo Yaqiong Wang Yaqiong Wang Jian Wang Jian Wang De-Wen Zhang De-Wen Zhang |
| author_facet |
Yao Meng Fangming Chen Mingrui Jiang Qin Guo Qin Guo Yaqiong Wang Yaqiong Wang Jian Wang Jian Wang De-Wen Zhang De-Wen Zhang |
| author_sort |
Yao Meng |
| title |
A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification |
| title_short |
A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification |
| title_full |
A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification |
| title_fullStr |
A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification |
| title_full_unstemmed |
A Homogeneous Label-Free Electrochemical microRNA Biosensor Coupling With G-Triplex/Methylene Blue Complex and λ-Exonuclease-Assisted Recycling Amplification |
| title_sort |
homogeneous label-free electrochemical microrna biosensor coupling with g-triplex/methylene blue complex and λ-exonuclease-assisted recycling amplification |
| publisher |
Frontiers Media S.A. |
| publishDate |
2021 |
| url |
https://doaj.org/article/feeccbd594c84535af99635f1f2159a7 |
| work_keys_str_mv |
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