RESUMO
The aim of the current study is to present a low-cost and easy-to-interpret colorimetric kit used to diagnose porcine circovirus 2 (PCV-2) to the naked eye, without any specific equipment. The aforementioned kit used as base hybrid nanoparticles resulting from the merge of surface active maghemite nanoparticles and gold nanoparticles, based on the deposition of specific PCV-2 antibodies on their surface through covalent bonds. In total, 10 negative and 40 positive samples (≥102 DNA copies/µL of serum) confirmed by qPCR technique were tested. PCV-1 virus, adenovirus, and parvovirus samples were tested as interferents to rule out likely false-positive results. Positive samples showed purple color when they were added to the complex, whereas negative samples showed red color; they were visible to the naked eye. The entire color-change process took place approximately 1 min after the analyzed samples were added to the complex. They were tested at different dilutions, namely pure, 1:10, 1:100, 1:1000, and 1:10,000. Localized surface plasmon resonance (LSPR) and transmission electron microscopy (TEM) images were generated to validate the experiment. This new real-time PCV-2 diagnostic methodology emerged as simple and economic alternative to traditional tests since the final price of the kit is USD 4.00.
RESUMO
Gold nanoparticles (AuNPs) can provide a simple, easy-to-use, inexpensive, at hand point-of-care (POC) fast to diagnose; however, AuNPs have the predisposition to form aggregations. Since the nanoparticles stability is an important issue, this article is aiming to study the long-term stability associated with the development of an immunosensor for clinical diagnosis. Here, we assessed two previous methods commonly described in the literature to prevent the formation of aggregate by studying pH and Tween® 20 (polysorbates) addition as surfactant. AuNPs were characterized through ultraviolet-visible (UV-Vis), dynamic light scattering (DLS) and transmission electron microscopy (TEM) and through analysis in the ImageJ software. We found that the Tween® 20 provided more than stable condition in aqueous solution in comparison to pH dependence. The fabricated AuNPs were further used to detect dengue virus and demonstrating that its use at pH 7.2 did not maintain reproducibility in the detection of dengue virus after one year. Unlike, the Tween® 20 modified AuNPs that detected dengue virus soon after the synthesis and over the course of one year demonstrating the high sensitivity of immunosensor. Finally, our result showed excellent dispersity throughout the year when using Tween® 20 to avoid aggregation.
