The rapid spread of the Coronavirus Disease 2019 (COVID-19) pandemic, caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pathogen has generated a huge international public health emergency. Currently the reference diagnostic technique for virus determination is Reverse Transcription Polymerase Chain Reaction (RT-PCR) real time analysis that requires specialized equipment, reagents and facilities and typically 3–4 h to perform. Thus, the realization of simple, low-cost, small-size, rapid and point-of-care diagnostics tests has become a global priority. In response to the current need for quick, highly sensitive and on-site detection of the SARS-CoV-2 virus in several aqueous solutions, a specific molecularly imprinted polymer (MIP) receptor has been designed, realized, and combined with an optical sensor. More specifically, the proof of concept of a SARS-CoV-2 sensor has been demonstrated by exploiting a plasmonic plastic optical fiber sensor coupled with a novel kind of synthetic MIP nano-layer, especially designed for the specific recognition of Subunit 1 of the SARS-CoV-2 Spike protein. First, we have tested the effectiveness of the developed MIP receptor to bind the Subunit 1 of the SARS-CoV-2 spike protein, then the results of preliminary tests on SARS-CoV-2 virions, performed on samples of nasopharyngeal (NP) swabs in universal transport medium (UTM) and physiological solution (0.9% NaCl), were compared with those obtained with RT-PCR. According to these preliminary results, the sensitivity of the proposed optical-chemical sensor proved to be higher than the RT-PCR one. Furthermore, a relatively fast response time (about 10 min) to the virus was obtained without the use of additional reagents.

Different lines of evidence indicate that monitoring the blood levels of therapeutic antibodies, characterized by high inter-individual variability, can help to optimize clinical decision making, improving patient outcomes and reducing costs with these expensive treatments. A surface plasmon resonance (SPR)-based immunoassay has recently been shown to allow highly reliable and robust monitoring of serum concentrations of infliximab, with significant advantages over classical ELISA. The next level of advancement would be the availability of compact and transportable SPR devices suitable for easy, fast and cheap point-of-care analysis. Here we report the data obtained with recently developed, cost-effective, optical-fibre-based SPR sensors (SPR-POF), which allow the construction of a compact miniaturized system for remote sensing. We carried out an extensive characterization of infliximab binding to an anti-infliximab antibody immobilized on the SPR-POF sensor surface. The present proof-of-principle studies demonstrate the feasibility of the proposed SPR-POF platform for the specific detection of infliximab, in both buffer and human serum, and pave the way for further technological improvements.

A specific aptameric sequence has been immobilized on short polyethyleneglycol (PEG) interface on gold nano-film deposited on a D-shaped plastic optical fiber (POFs) probe, and the protein binding has been monitored exploiting the very sensitive surface plasmon resonance (SPR) phenomenon. The receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein has been specifically used to develop an aptasensor. Surface analysis techniques coupled to fluorescence microscopy and plasmonic analysis have been utilized to characterize the biointerface. Spanning a wide protein range (25÷1000 nM), the SARS-Cov-2 spike protein was detected with a Limit of Detection (LoD) of about 37 nM. Different interferents (BSA, AH1N1 hemagglutinin protein and MERS spike protein) have been tested confirming the specificity of our aptasensor. Finally, a preliminary test in diluted human serum encouraged its application in a point-of-care device, since POF-based aptasensor represent a potentially low-cost compact biosensor, characterized by a rapid response, a small size and could be an ideal laboratory portable diagnostic tool.

In this work, we present a novel biochemical sensing approach based on a plasmonic sensor chip, combined with a specific receptor, excited and interrogated via a custom 3D-printed holder through a transmission-based experimental setup, exploiting polymer optical fibers. The setup is designed to measure a disposable plasmonic chip based on a gold nanograting fabricated on a polymethylmethacrylate substrate. The examined sensor configurations here presented are simulated, realized, and experimentally tested. More specifically, first, a numerical analysis is carried out by changing several sensor parameters, then an experimental optical characterization of different sensor configurations is reported. Finally, to test the biosensing capabilities of the proposed method, as a proof of concept, we deposit on the best sensor configuration a biomimetic receptor specific for bovine serum albumin detection. The experimental results demonstrate that the proposed sensor shows an ultra-low limit of detection, equal to about 37 pmol/L.

Abstract:In this paper, we show that light-diffusing fibers (LDF) can be efficiently used as host material for surface plasmon resonance (SPR)-based refractive index sensing. This novel platform does not require a chemical procedure to remove the cladding or enhance the evanescent field, which is expected to give better reproducibility of the sensing interface. The SPR sensor has been realized by first removing the cladding with a simple mechanical stripper, and then covering the unclad fiber surface with a thin gold film. The tests have been carried out using water-glycerin mixtures with refractive indices ranging from 1.332 to 1.394. The experimental results reveal a high sensitivity of the SPR wavelength to the outer medium's refractive index, with values ranging from 1500 to~4000 nm/RIU in the analyzed range. The results suggest that the proposed optical fiber sensor platform could be used in biochemical applications.

A novel Molecularly Imprinted Polymer (MIP) able to bind perfluorinated compounds, combined with a surface plasmon resonance (SPR) optical fiber platform, is presented. The new MIP receptor has been deposited on a D-shaped plastic optical fiber (POF) covered with a photoresist buffer layer and a thin gold film. The experimental results have shown that the developed SPR-POF-MIP sensor makes it possible to selectively detect the above compounds. In this work, we present the results obtained with perfluorooctanoate (PFOA) compound, and they hold true when obtained with a perfluorinated alkylated substances (PFAs) mixture sample. The sensor’s response is the same for PFOA, perfluorooctanesulfonate (PFOS) or PFA contaminants in the C4–C11 range. We have also tested a sensor based on a non-imprinted polymer (NIP) on the same SPR in a D-shaped POF platform. The limit of detection (LOD) of the developed chemical sensor was 0.13 ppb. It is similar to the one obtained by the configuration based on a specific antibody for PFOA/PFOS exploiting the same SPR-POF platform, already reported in literature. The advantage of an MIP receptor is that it presents a better stability out of the native environment, very good reproducibility, low cost and, furthermore, it can be directly deposited on the gold layer, without modifying the metal surface by functionalizing procedures.

The development of optical biosensors for the rapid and costless determination of clinical biomarkers is of paramount importance in medicine. Here we report a fast and low-cost biosensor based on a plasmonic D-shaped plastic optical fibre (POF) sensor derivatized with an aptamer specific for the recognition of thrombin, the target marker of blood homeostasis and coagulation cascade. In particular, we designed a functional interface based on a Self Assembled Monolayer (SAM) composed of short Poly Ethylene Glycol (PEG) chains and biotin-modified PEG thiol in ratio 8:2 mol:mol, these latter serving as baits for the binding of the aptamer through streptavidin-chemistry. The SAM was studied by X-ray Photoelectron Spectroscopy (XPS) analysis, static contact angle (CA), Surface Plasmon Resonance (SPR) in POFs, and fluorescence microscopy on gold surface. The optimized SAM composition enabled the immobilization of about 112 ng/cm2 of aptamer. The thrombin detection exploiting POF-Aptasensor occurred in short times (5–10 minutes), the reached Limit of Detection (LOD) was about 1 nM, and the detection range was 1.6–60 nM, indicating the POF-Aptasensor well addresses the needs for a low-cost, simple to use and to realize, rapid, small size and portable diagnostic platform.

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