Binding of pDNA with cDNA using hybridization strategy towards monitoring of Haemophilus influenza genome in human plasma samples
Language English Country Netherlands Media print-electronic
Document type Journal Article
PubMed
32057844
DOI
10.1016/j.ijbiomac.2020.02.062
PII: S0141-8130(19)40372-3
Knihovny.cz E-resources
- Keywords
- Bioassay, DNA hybridization, Haemophilus influenza, Nucleic acid binding, Silver nanoparticles,
- MeSH
- Biosensing Techniques MeSH
- Electrochemical Techniques MeSH
- Genome, Bacterial * MeSH
- Haemophilus influenzae genetics MeSH
- Nucleic Acid Hybridization methods MeSH
- DNA, Complementary * MeSH
- Metal Nanoparticles ultrastructure MeSH
- Quantum Dots MeSH
- Humans MeSH
- Reproducibility of Results MeSH
- Cell-Free Nucleic Acids * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- DNA, Complementary * MeSH
- Cell-Free Nucleic Acids * MeSH
Haemophilus Influenza leads to respiratory infections such as sinusitis, acute otitis media, pneumonia and bronchitis. In addition, it causes invasive infections such as cellulite, septic arthritis, and meningitis. Therefore, quick and sensitive detection of H. influenza is of great importance in medical microbiology. In this study, a novel DNA-based bioassay was developed to the monitoring of Haemophilus influenza genome in human plasma samples using binding of pDNA with cDNA. DNA hybridization strategy was used to investigation of DNAs binding. For this purpose, silver nanoparticle doped graphene quantum dots inks functionalized by D-penicillamine (Ag NPs-DPA-GQDs) were synthesized and deposited on the surface of glass carbon electrode (GCE). Also, gold nanoparticles functionalized with cysteamine (CysA-AuNPs) were deposited on the surface of the Ag-DPA-GQDs modified GCE. Afterward, thiolated DNA probe was immobilized on the surface of the modified electrode. DNA hybridization was monitored using square wave voltammetry (SWV) technique. Engineered genosensor indicated good performance with high specificity and sensitivity for detection of Haemophilus influenza genome. Under optimal conditions, linear range and low limit of quantitation (LLOQ) were obtained as target concentrations ranging from 1 pM-1 ZM and 1 ZM, respectively. The designed biosensor also showed high capability of discriminating one-base, two-base and three-base mismatched sequences. Also, the prepared genosensor could be easily regenerated and reused to evaluate hybridization process.
Department of Nanotechnology Faculty of Science and Chemistry Urmia University Urmia Iran
Pharmaceutical Analysis Research Center Tabriz University of Medical Sciences Tabriz Iran
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