RESUMO
Here, the construction and characterization of the first immunosensor for highly sensitive and label free detection of Fig mosaic virus (FMV) is reported. The specific antibody against nucleocapsid of the virus was raised and immobilized at the surface of 11-mercaptoundecanoic acid (MUA) and 3-mercapto propionic acid (MPA) modified gold electrode, via carbodiimide coupling reaction. The immunosensor fabrication steps were characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The electrochemical detection of FMV was conducted using differential pulse voltammetry in ferri/ferrocyanide solution as a redox probe. The proposed immunosensor exhibited high selectivity, good reproducibility and high sensitivity for FMV detection in a range from 0.1â¯nM to 1⯵M with a detection limit of 0.03â¯nM. Moreover, good results were obtained for determination of FMV in real samples, indicating the feasibility of the developed immunosensor for detection of fig mosaic disease, without the need for molecular (e.g. PCR) amplification.
Assuntos
Técnicas Biossensoriais/métodos , Espectroscopia Dielétrica/métodos , Ficus/virologia , Imunoensaio/métodos , Vírus de Plantas/isolamento & purificação , Ácido 3-Mercaptopropiônico , Anticorpos Imobilizados/química , Técnicas Eletroquímicas/métodos , Eletrodos , Álcoois Graxos/química , Ouro/química , Limite de Detecção , Oxirredução , Vírus de Plantas/química , Vírus de Plantas/imunologia , Compostos de Sulfidrila/químicaRESUMO
For the first time a highly sensitive and selective fluorimetric optode for determination of trace amounts of Ho(3+) ions was prepared. The sensing system was prepared by incorporating of N,N-bis(salicylidene)-naphthylene-1,8-diamine (L) as a neutral Ho(3+)-selective fluoroionophore, in a plasticized PVC membrane containing sodium tetraphenyl borate as a lipophilic anionic additive. The response of the sensor is based on the strong fluorescence quenching of L by Ho(3+) ions. At pH 5.4, the proposed sensor displays a calibration curve over a wide concentration range of 1.0×10(-10)-1.0×10(-3)M, with a relatively fast response time of less than 1 min. In addition to high stability, high reproducibility and a relatively long working lifetime, the sensor shows a good selectivity towards Ho(3+) ion with respect to common coexisting cations. The fluorescence optode was applied to determination of holmium ion contents of water samples.