Nanostructured sensor platform based on organic polymer conjugated to metallic nanoparticle for the impedimetric detection of SARS-CoV-2 at various stages of viral infection.

The projection of new biosensing technologies for genetic identification of SARS-COV-2 is essential in the face of a pandemic scenario. For this reason, the current research aims to develop a label-free flexible biodevice applicable to COVID-19. A nanostructured platform made of polypyrrole (PPy) and gold nanoparticles (GNP) was designed for interfacing the electrochemical signal in miniaturized electrodes of tin-doped indium oxide (ITO). Oligonucleotide primer was chemically immobilized on the flexible transducers for the biorecognition of the nucleocapsid protein (N) gene. Methodological protocols based on cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and atomic force microscopy (AFM) were used to characterize the nanotechnological apparatus. The biosensor's electrochemical performance was evaluated using the SARS-CoV-2 genome and biological samples of cDNA from patients infected with retrovirus at various disease stages. It is inferred that the analytical tool was able to distinguish the expression of SARS-CoV-2 in patients diagnosed with COVID-19 in the early, intermediate and late stages. The biosensor exhibited high selectivity by not recognizing the biological target in samples from patients not infected with SARS-CoV-2. The proposed sensor obtained a linear response range estimated from 800 to 4000 copies µL-1 with a regression coefficient of 0.99, and a detection limit of 258.01 copies µL-1. Therefore, the electrochemical biosensor based on flexible electrode technology represents a promising trend for sensitive molecular analysis of etiologic agent with fast and simple operationalization. In addition to early genetic diagnosis, the biomolecular assay may help to monitor the progression of COVID-19 infection in a novel manner.

Label-free nanostructured biosensor for Schistosoma mansoni detection in complex biological fluids.

Schistosomiasis is a neglected tropical disease with a worldwide prevalence. Neuroschistosomiasis is the most severe presentation of the disease and affects the central nervous system. In this work, Schistosoma mansoni detection was based on self-assembled layers of 3-mercaptopropyltrimethoxysilane (MPTS) and electrosynthesized gold nanoparticles (AuNPs). The DNA probe was chemisorbed onto AuNPs. The biosensor was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The impedimetric response of the MPTS-AuNPs-DNAprobe system indicates an effective modification of the electrode surface. Topographical atomic force microscopy images were used to characterize the self-assembled layers on the gold electrode surface. The proposed biosystem was able to recognize the S. mansoni genome sequence at different concentrations in samples of urine, cerebrospinal fluid, and serum. Several concentration ranges were evaluated: urine (27-50 pg µL-1), cerebrospinal fluid (25-60 pg µL-1), and serum (27-42 pg µL-1). The limit detection (LOD) of the biosensor was 0.6 pg µL-1. The developed label-free genosensor was able to detect small concentrations of S. mansoni DNA in complex biological fluids.

Impedimetric nanostructured genosensor for detection of schistosomiasis in cerebrospinal fluid and serum samples.

Schistosomiasis is a neglected disease closely related to the low levels of social development and a serious public health problem. In this work, we performed an electrochemical detection of Schistosoma mansoni DNA with a self-assembled monolayer of mercaptobenzoic acid (MBA) immobilizing nanostructures composed of gold nanoparticles (AuNPs) and magnetite nanoparticles (Fe3O4_NPs). Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to monitor the hybridization process. MBA-Fe3O4_NPs-AuNPs-DNAprobe system reveals an effective electrochemical response indicating the surface modification. The proposed biosystem was capable to recognize specific nucleotide sequence of S. mansoni present in cerebrospinal fluid (CFS) and serum samples at different genome DNA concentrations. The biorecognition resulted in an increase in the electron transfer resistance and a decrease of the current peaks at higher DNA concentrations during electrochemical measurements. The developed platform showed a DNA detection limit of 0.781 and 0.685pgµL-1 for serum and CFS, respectively. Therefore, the obtained biosensor can be considered as a useful tool for specific detection of S. mansoni at low concentrations in various biological fluids.