Fluorometric determination of ssDNA based on functionalized magnetic microparticles and DNA supersandwich self-assemblies
| Autor: | Suping Bai, Hai-Xia Lv, Yufei Liu, Ningning He, Jingfang Shangguan, Qianqian Wang |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Streptavidin
Concatemer Aptamer Biotin DNA Single-Stranded Diamines Fluorescence spectroscopy Analytical Chemistry chemistry.chemical_compound Blood serum Adenosine Triphosphate Limit of Detection Humans Fluorometry Benzothiazoles Organic Chemicals Chemistry Magnetic Phenomena Nucleic Acid Hybridization Aptamers Nucleotide Ebolavirus Combinatorial chemistry DNA Concatenated Intercalating Agents DNA Viral Nucleic acid SYBR Green I Quinolines DNA Probes DNA |
| Zdroj: | Mikrochimica acta. 186(11) |
| ISSN: | 1436-5073 |
| Popis: | A method is described for the determination of DNA via nucleic acid amplification by using nucleic acid concatemers that result from DNA supersandwich self-assemblies (SSAs). The method employs two auxiliary probes to form self-assembled biotin SSAs. These exhibit strong fluorescence if labeled with intercalator SYBR Green I. In the presence of the target (as exemplified for a 30-mer), streptavidin is released from the surface of the functionalized magnetic microparticles (FMMPs) by competitive hybridization on the surface. However, the SSA products do not conjugate to the FMMPs. This leads to a large amount of SYBR Green I intercalated into the concatemers and eventually results in amplified fluorescence in the supernate. The SSA products can be prepared beforehand, and amplification therefore can be completed within 50 min. The method is more efficient than any other conventional amplification. The detection limit for the 30-mer is 26.4 fM which is better by a factor of 10 compared to other amplification methods. Conceivably, the method can be further extended to the determination of a wide variety of targets simply by replacing the sequences of the probes. Finally, this rapid and highly sensitive method was employed for detection of Ebola virus gene (≈30-mer) and ATP in spiked serum with satisfactory results. Graphical abstract A high sensitivity and efficiency bioassay is described based on functionalized magnetic microparticles and DNA supersandwich self-assemblies. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |