DNA-only Cascade: A Universal Tool for Signal Amplification, Enhancing the Detection of Target Analytes
| Autor: | Nicole E. Lima, Nicole J. Hasick, Simon M. Bone, Alison V. Todd, Elisa Mokany, Simon Erskine |
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| Rok vydání: | 2014 |
| Předmět: |
DNA
Bacterial Analyte Deoxyribozyme Neisseria meningitidis Polymorphism Single Nucleotide Analytical Chemistry Nucleic acid thermodynamics chemistry.chemical_compound Adenosine Triphosphate Bacterial Proteins Deoxyadenosine triphosphate Humans Nucleotide Fluorescent Dyes Ions chemistry.chemical_classification Nucleic Acid Hybridization Streptococcus DNA DNA Catalytic Nucleic acid amplification technique Combinatorial chemistry Small molecule Meningococcal Infections Lead Biochemistry chemistry Nucleic acid Nucleic Acid Amplification Techniques |
| Zdroj: | Analytical Chemistry. 86:9106-9113 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | Diagnostic tests performed in the field or at the site of patient care would benefit from using a combination of inexpensive, stable chemical reagents and simple instrumentation. Here, we have developed a universal "DNA-only Cascade" (DoC) to quantitatively detect target analytes with increased speed. The DoC utilizes quasi-circular structures consisting of temporarily inactivated deoxyribozymes (DNAzymes). The catalytic activity of the DNAzymes is restored in a universal manner in response to a broad range of environmental and biological targets. The present study demonstrates DNAzyme activation in the presence of metal ions (Pb(2+)), small molecules (deoxyadenosine triphosphate) and nucleic acids homologous to genes from Meningitis-causing bacteria. Furthermore, DoC efficiently discriminates nucleic acid targets differing by a single nucleotide. When detection of analytes is orchestrated by functional nucleic acids, the inclusion of DoC reagents substantially decreases time for detection and allows analyte quantification. The detection of nucleic acids using DoC was further characterized for its capability to be multiplexed and retain its functionality following long-term exposure to ambient temperatures and in a background of complex medium (human serum). |
| Databáze: | OpenAIRE |
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