A highly sensitive quantum dots-DNA nanobiosensor based on fluorescence resonance energy transfer for rapid detection of nanomolar amounts of human papillomavirus 18

[Display omitted] •Design of a highly selective and sensitive FRET-based nanobiosensor for low level detection of HPV18.•Use of TGA-CdTe quantum dots as donor and fluorophore Cy5-labeled oligonucleotides as acceptor.•Obtaining a linear range of 1–50nM with a detection limit of 0.2nM.•Successful appl...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Journal of pharmaceutical and biomedical analysis 2017-03, Vol.136, p.140-147
Hauptverfasser: Shamsipur, Mojtaba, Nasirian, Vahid, Mansouri, Kamran, Barati, Ali, Veisi-Raygani, Asad, Kashanian, Soheila
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Beschreibung
Zusammenfassung:[Display omitted] •Design of a highly selective and sensitive FRET-based nanobiosensor for low level detection of HPV18.•Use of TGA-CdTe quantum dots as donor and fluorophore Cy5-labeled oligonucleotides as acceptor.•Obtaining a linear range of 1–50nM with a detection limit of 0.2nM.•Successful application of the nanobiosensor to analysis of derived nucleotides from HPV18 patients. A very sensitive and convenient nanobiosensor based on fluorescence resonance energy transfer (FRET) was developed for the detection of a 22-mer oligonucleotides sequence in Human Papillomavirus 18 virus (HPV18) gene. For this purpose, water-soluble CdTe quantum dots (QDs) were synthesized and, subsequently, amino-modified 11-mer oligonucleotide as one of the two necessary probes was attached to QDs surface to form functional QDs-DNA conjugates. Right after addition of the QDs-DNA and a second Cyanine5 (Cy5)-labeled 11-mer oligonucleotide probe to the DNA target solution, the sandwiched hybrids were formed. The resulting hybridization brings the Cy5 fluorophore as the acceptor to close proximity of the QDs as donor, so that an effective transfer of energy from the excited QDs to the Cy5 probe would occur via FRET processing. The fluorescence intensity of Cy5 found to linearly enhance by increasing the DNA target concentration from 1.0 to 50.0nM, with a detection limit of 0.2nM. This homogeneous DNA detection method does not require excessive washing and separation steps of un-hybridized DNA, due to the fact that no FRET can be observed when the probes are not ligated. Finally, feasibility and selectivity of the proposed one-spot DNA detection nanobiosensor were investigated by analysis of derived nucleotides from HPV18 and mismatched sequences.
ISSN:0731-7085
1873-264X
DOI:10.1016/j.jpba.2017.01.002