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1.
Int J Biol Macromol ; 112: 364-370, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29378271

RESUMO

Staphylococcus aureus (S. aureus) is a pathogenic bacteria which causes infectious diseases and food poisoning. Current diagnostic methods for infectious disease require sophisticated instruments, long analysis time and expensive reagents which restrict their application in resource-limited settings. Electrochemical paper based analytical device (EPAD) was developed by integrating graphene nano dots (GNDs) and zeolite (Zeo) using specific DNA probe. The ssDNA/GNDs-Zeo modified paper based analytical device (PAD) was characterized using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The genosensor was optimized at pH7.4 and incubation temperature of 30°C. A linear current response with respect to target DNA concentrations was obtained. The limit of detection (LOD) of the proposed sensor was found out to be 0.1nM. The specificity was confirmed by introducing non-complimentary target DNA to ssDNA/GNDs-Zeo modified PAD. The suitability of the proposed EPAD genosensor was demonstrated with fruit juice samples mixed with S. aureus. The proposed EPAD genosensor is a low cost, highly specific, easy to fabricate diagnostic device for detection of S. aureus bacteria which requires very low sample volume and minimum analysis time of 10s.


Assuntos
DNA Bacteriano/isolamento & purificação , Grafite/química , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus/isolamento & purificação , DNA Bacteriano/genética , Técnicas Eletroquímicas , Humanos , Papel , Pontos Quânticos/química , Infecções Estafilocócicas/genética , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidade , Zeolitas/química
2.
Biosens Bioelectron ; 86: 566-574, 2016 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-27448547

RESUMO

Nanocrystals of zeolites (Nanocrys Zeo) and Multi-walled carbon nanotubes (MWCNT) based diagnostic genosensor was employed for detection of polymerase chain (PCR) amplified HBVDNA in blood of hepatitis B patients. The ssDNA-nanocomposite modified electrode was characterized by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The hybridization between ss DNA probe and target ss DNA was detected by reduction in current, generated by interaction of methylene blue (MB) with free guanine (3'G) of ssDNA. Nanocrys zeo were deposited on the Fluorine doped tin oxide glass electrode (FTO) by drop-casting method for better immobilization of ss DNA while MWCNTs are incorporated into the zeolite-assembly to enhance the electro-conductivity of the present genosensor. The ssDNA-nanocomposite modified FTO electrode exhibited optimum current within 5s, at pH 5.6, and incubation temperature of 45°C. The value of charge transfer resistance (Rct) was linear with the number of copies of target DNA between 150 and 10(6) copies/ml. The limit of detection (LOD) of the sensor was 50 copies/ml. Within and between batches coefficients of variation (CV) were 2.5% and 3.2% respectively. Results obtained with our genosensor were also correlated with those by RT-PCR and r(2) value found with good accuracy of 97%. The electrode was reused by dipping it into 0.1M NaOH for 3min and lost 50% of its initial activity in 4 weeks. Furthermore the technique employed for detection of HBV is EIS, which is convenient and required less analysis time.


Assuntos
DNA Viral/sangue , Espectroscopia Dielétrica/métodos , Técnicas Eletroquímicas/métodos , Vírus da Hepatite B/isolamento & purificação , Hepatite B/sangue , Nanotubos de Carbono/química , Zeolitas/química , Sondas de DNA/química , DNA de Cadeia Simples/química , DNA Viral/análise , DNA Viral/genética , DNA Viral/isolamento & purificação , Impedância Elétrica , Hepatite B/diagnóstico , Hepatite B/virologia , Vírus da Hepatite B/genética , Humanos , Limite de Detecção , Nanocompostos/química , Nanotubos de Carbono/ultraestrutura
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