RESUMO
Zika virus (ZIKV) infection is associated with the Guillain-Barré syndrome, and when non-vector congenital transmission occurs, fetal brain abnormalities are expected. After ZIKV infection, the blood, breast milk, and other body fluids contain low viral loads. Their detection is challenging as it requires the processing of larger input volumes of the clinical samples. Pre-enrichment is a valuable strategy to increase the analyte concentration. Therefore, the authors propose the use of a hierarchal composite polyaniline-(electrospun nanofiber) hydrogel mat (ENM) for the simultaneous enrichment and impedimetric sensing of ZIKV viral particles. The electrospinning conditions of polyvinyl alcohol and alginate, including blend formulation, were optimized through a factorial design. Disintegration and gelatinization were controlled via cross-linking to improve the hydrogel properties. Hierarchization was achieved by in situ chemical deposition of conductive polyaniline. The carboxyl groups of the ENM were used for the covalent immobilization of anti-ZIKV polyclonal antibodies used in the specific recognition of ZIKV within the medium of Vero cell culture. The specific capture and desorption of virions were studied at different pHs. ENMs were characterized by scanning electron microscopy and FTIR. Atomic force microscopy along with UV-vis and electrochemical impedance spectroscopies was used to monitor the antibody immobilization, ZIKV capture, and elution processes. Our results show that 14.2 mg (0.25 cm3) of ENM can capture 38.7 ± 2.5 µg of ZIKV with a desorption rate of 99.97% (38.29 ± 2.7 µg ZIKV), which is reusable for at least three times. Therefore, the capture capacity (micrograms of ZIKV captured per milligram of ENM) of polyaniline-hierarchized mats was 2.72 µg ZIKV/mg. The impedance LOD value was determined to be 2.76 µg of ZIKV particles (approximately 6.6 × 103 PFU/mL). As a result, we present a fast small-scale purification system that can simultaneously monitor ZIKV electrochemically and optically.
Assuntos
Alginatos/química , Compostos de Anilina/química , Técnicas Biossensoriais/métodos , Nanofibras/química , Carga Viral/métodos , Zika virus/isolamento & purificação , Animais , Anticorpos Imobilizados/imunologia , Anticorpos Antivirais/imunologia , Sangue/virologia , Chlorocebus aethiops , Técnicas Eletroquímicas , Hidrogéis/química , Imunoensaio/métodos , Limite de Detecção , Células Vero , Zika virus/imunologiaRESUMO
A generalization of the concept of multimode interference sensors is presented here for the first time, to the best of our knowledge. The existing bimodal and trimodal sensors correspond to particular cases of those interference sensors. A thorough study of the properties of the multimode waveguide section provided a deeper insight into the behavior of this class of sensors, which allowed us to establish new criteria for designing more sensitive structures. Other challenges of using high-order modes within the sensing area of the device reside in the excitation of these modes and the interpretation of the output signal. To overcome these, we developed a novel structure to excite any desired high-order mode along with the fundamental mode within the sensing section, while maintaining a fine control over the power distribution between them. A new strategy to detect and interpret the output signal is also presented in detail. Finally, we designed a high-order sensor for which numerical simulations showed a theoretical limit of detection of 1.9×10-7 RIU, making this device the most sensitive multimode interference sensor reported so far.