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1.
Anal Chim Acta ; 905: 134-9, 2016 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-26755147

RESUMO

A mass sensitive quartz crystal microbalance (QCM) based genosensor has been developed using breast cancer 1 (BRCA1) gene as a model gene. We modified the traditional sandwich assay by conjugating reporter probe DNA (DNA-r) with an assembly of gold nanoparticles leading to an increased mass on the surface, which enhanced the sensitivity to few orders of magnitude. The unique cleavage function of endonuclease is used for achieving the selectivity to complementary DNA over mismatched DNA. With this combination, the sensor exhibited excellent sensitivity with a detection limit of 10 aM BRCA1 gene and it showed good selectivity for even single base mismatch DNA targets. This ultrasensitive and cost-effective DNA detection protocol can be extended to the direct analysis of any non-amplified genomic DNA.


Assuntos
DNA/análise , Técnicas de Microbalança de Cristal de Quartzo , Microscopia Eletrônica de Varredura
2.
Biosens Bioelectron ; 65: 333-40, 2015 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-25461178

RESUMO

In this work, we report a simple strategy for signal amplification using appropriately functionalized gold nanoparticles in an electrochemical genosensor which led to attomolar detection of breast cancer 1 (BRCA1) gene. The sensor was developed by the layer-by-layer assembly of mercaptopropionic acid (MPA), polyethylene glycol (PEG) functionalized gold nanoparticle (AuNPPEG), capture DNA (DNA-c), target BRCA1 DNA (DNA-t) and gold nanoparticle labeled reporter DNA (DNA-r.AuNP) on gold electrode. PEG functionalized gold nanoparticles on the MPA surface provided good electron conducting path nullifying the insulating effect of MPA and also act as a proper immobilization platform for the DNA-c by the large number of carboxyl groups present on the functionalized gold nanoparticles. We demonstrated that the incorporation of MPA functionalized gold nanoparticles (AuNPMPA) as an electrochemical label in this sensor design could significantly enhance the sensitivity in the detection. The DNA hybridization of DNA-r.AuNP with target probe was measured by chronoamperometry, electrochemical impedance spectroscopy (EIS), and scanning tunnelling spectroscopy (STS). Electrochemical quartz crystal microbalance (EQCM) experiments were used to support the detection and also to calculate the number of adsorbed molecules on the surface. Under optimum conditions the present sensor exhibited high sensitivity and a very low detection limit of 50attomolar DNA target (294.8attogram BRCA1gene/ml). It shows excellent selectivity against non complementary sequences and 3 base mismatch complementary targets. It also shows good reproducibility, stability and reusability and the developed sensor surface is suitable for point-of care applications.


Assuntos
Técnicas Biossensoriais/métodos , DNA/análise , Genes BRCA1 , Ouro/química , Nanopartículas Metálicas/química , Hibridização de Ácido Nucleico/métodos , DNA/genética , Espectroscopia Dielétrica/métodos , Técnicas Eletroquímicas/métodos , Eletrodos , Humanos , Limite de Detecção , Polietilenoglicóis/química , Técnicas de Microbalança de Cristal de Quartzo/métodos , Reprodutibilidade dos Testes
3.
J Colloid Interface Sci ; 428: 214-21, 2014 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-24910056

RESUMO

We report a simple method for decorating carboxyl functionalized few-layer graphene with titania (TiO2) nanoparticles by sonication and stirring under room temperature. The nanocomposites showed a remarkable improvement in visible light driven photocatalysis. From Raman and XRD analysis the number of layers of graphene was found to be 3. The TiO2 decorated few-layer graphene (FLG) sheets were characterized by electron microscopy, Raman spectroscopy, infrared spectroscopy, XRD and UV-vis spectroscopy. Titania nanoparticles were uniformly decorated on FLG matrix. The incorporation of titania on FLG enhanced the visible light photocatalytic activity of titania, lowered the electron hole recombination and improved the electron hole mobility. The enhanced life time of the charge carriers was confirmed from the photocurrent measurements. Compared to bare TiO2 nanoparticles the FLG-TiO2 nanocomposites exhibited rapid degradation of Rhodamine B (Rhd B) under solar radiation. It was found that adsorption of dye molecules and the rate of degradation have been greatly enhanced in the FLG decorated with TiO2. The rapid degradation of Rhd B using carboxyl functionalized FLG-TiO2 within 8 min under solar radiation and 20 min under 30 W UV tube with very low concentration (0.01 wt.%) of the photocatalyst is the highlight of the present report. The mechanism of degradation and charge separation ability of the nanocomposite are also explored.

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