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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Yu, Ruqin Jiang, Jianhui Sun, Chenhu Zhang, Liangliang Shen, Guoli |
| Description | Author Affiliation: Sun C ( State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.) |
| Abstract | This paper describes a novel electrochemical DNA sensor for the simple, sensitive and specific detection of nucleic acids based on proximity-dependent DNA ligation assays with the DNAzyme amplification of hairpin substrate signal. A long DNA strand contains the catalytic motif of Mg(2+)-dependent 10-23 DNAzyme, acting as the recognition probe. When the target DNA was introduced into the system, part of it was complementary to 5'-end of the recognition probe, resulting in the ligation of a stable duplex, the unbinded part of the target DNA was acted as one binding arm for the DNAzyme. This duplex containing a complete 10-23 DNAzyme structure could cleave the purine-pyrimidine cleavage site of the hairpin substrate, which resulted in the fragmentation of the hairpin structure and the release of two single-stranded nucleic acids, one of which was biotinylated and acted as the signal probe. An immobilized thiolated capture probe could bind with the signal probe, using biotin as a tracer in the signal probe, and streptavidin-alkaline phosphatase (SA-ALP) as reporter molecule. The activity of the immobilized enzyme was voltammetrically determined by measuring the amount of 1-naphthol generated after 5 min of enzymatic dephosphorylation of 1-naphthyl phosphate. The results revealed that the sensor showed a sensitive response to complementary target sequences of H. pylori in a concentration range from 100 fM to 1 nM, with a detection limit of 50 fM. In addition, the sensing system could discriminate the complementary sequence from mismatched sequences, with high sensitivity and reusability. |
| ISSN | 09565663 |
| Issue Number | 11 |
| Volume Number | 25 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2010-07-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Biosensing Techniques Instrumentation Conductometry Dna Analysis Genetics Ligase Chain Reaction Nucleic Acid Amplification Techniques Dna, Catalytic Chemistry Equipment Design Equipment Failure Analysis Reproducibility Of Results Sensitivity And Specificity Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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