How Nanophotonic Label-Free Biosensors Can Contribute to Rapid and Massive Diagnostics of Respiratory Virus Infections : COVID-19 Case

Altres ajuts: Generalitat de Catalunya /CERCAThis ACS article is provided to You under the terms of this Standard ACS AuthorChoice License. License: https://pubs.acs.org/page/policy/authorchoice_termsofuse.htmlThe global sanitary crisis caused by the emergence of the respiratory virus SARS-CoV-2 and the COVID-19 outbreak has revealed the urgent need for rapid, accurate, and affordable diagnostic tests to broadly and massively monitor the population in order to properly manage and control the spread of the pandemic. Current diagnostic techniques essentially rely on polymerase chain reaction (PCR) tests, which provide the required sensitivity and specificity. However, its relatively long time-to-result, including sample transport to a specialized laboratory, delays massive detection. Rapid lateral flow tests (both antigen and serological tests) are a remarkable alternative for rapid point-of-care diagnostics, but they exhibit critical limitations as they do not always achieve the required sensitivity for reliable diagnostics and surveillance. Next-generation diagnostic tools capable of overcoming all the above limitations are in demand, and optical biosensors are an excellent option to surpass such critical issues. Label-free nanophotonic biosensors offer high sensitivity and operational robustness with an enormous potential for integration in compact autonomous devices to be delivered out-of-the-lab at the point-of-care (POC). Taking the current COVID-19 pandemic as a critical case scenario, we provide an overview of the diagnostic techniques for respiratory viruses and analyze how nanophotonic biosensors can contribute to improving such diagnostics. We review the ongoing published work using this biosensor technology for intact virus detection, nucleic acid detection or serological tests, and the key factors for bringing nanophotonic POC biosensors to accurate and effective COVID-19 diagnosis on the short term

Optical nanogap antennas as plasmonic biosensors for the detection of miRNA biomarkers

Nanoplasmonic biosensors based on nanogap antenna structures usually demand complex and expensive fabrication processes in order to achieve a good performance and sensitive detection. We here report the fabrication of large-area nanoplasmonic sensor chips based on nanogap antennas by employing a customized, simple and low-cost colloidal lithography process. By precisely controlling the angle for tilted e-beam metal evaporation, an elliptical mask is produced, which defines the total length of the dipole antenna nanostructures while assuring that the plasmonic response is oriented in the same direction along the sensor chip. Large-area sensor chips of nanogap antennas formed by pairs of gold nanodisks separated by gaps with an average size of 11.6 ± 4.7 nm are obtained. The optical characterization of the nanogap antenna structures in an attenuated total reflection (ATR) configuration shows a bulk refractive index sensitivity of 422 nm per RIU, which is in agreement with FDTD numerical simulations. The biosensing potential of the cm-sized nanostructured plasmonic sensor chips has been evaluated for the detection of miRNA-210, a relevant biomarker for lung cancer diagnosis, through a DNA/miRNA hybridization assay. A limit of detection (LOD) of 0.78 nM (5.1 ng mL) was achieved with no need of further amplification steps, demonstrating the high sensitivity of these plasmonic nanogap antennas for the direct and label-free detection of low molecular weight biomolecules such as miRNAs

Introduction to the special issue on ‘‘hydro-meteorological time series analysis and their relation to climate change’’

Peer ReviewedPostprint (author's final draft

Influence of Food Matrices and the Population of Interfering Microorganisms on the Determination of Listeria monocytogenes by Conventional Methods and VIDAS

In this study, the possible influence of the food matrix and the interfering population of microorganisms on the detection and count of Listeria monocytogenes in three common foods of the Spanish diet (Spanish omelette, fresh cheese and vegetable salad) was determined. Four groups were assayed: one control, two groups with interfering microorganisms (Salmonella Enteritidis, Staphylococcus aureus and Proteus mirabilis) with different levels of L. monocytogenes and a final group only contaminated with L. monocytogenes. The samples were analyzed with the normalized method (UNE-EN ISO 11290:2018) and with an alternative technique (VIDAS). The results show that the presence of interfering microorganisms did not seem to interfere with the determination of L. monocytogenes. Furthermore, the type of food did not seem to influence the determination of L. monocytogenes, but the culture media used showed differences. In fact, regardless of the type of food, the ALOA medium showed higher sensitivity than the other media, with higher recovery in 100% of samples (only for the Spanish omelette in Group B was the result the same as that for PALCAM, −8.11 log cfu/g). The results obtained using the VIDAS were not influenced by any of the factors or conditions used and show 100% efficiency

Real-time monitoring of fenitrothion in water samples using a silicon nanophotonic biosensor

[EN] Due to the large quantities of pesticides extensively used and their impact on the environment and human health, a prompt and reliable sensing technique could constitute an excellent tool for in-situ monitoring. With this aim, we have applied a highly sensitive photonic biosensor based on a bimodal waveguide interferometer (BiMW) for the rapid, label-free, and speci¿c quanti¿cation of fenitrothion (FN) directly in tap water samples. After an optimization protocol, the biosensor achieved a limit of detection (LOD) of 0.29 ng mL¿¿1 (1.05 nM) and a half-maximal inhibitory concentration (IC50)of 1.71 ng mL¿¿1 (6.09 nM) using a competitive immunoassay and employing diluted tap water. Moreover, the biosensor was successfully employed to determine FN concentration in blind tap water samples obtaining excellent recovery percentages with a time-to-result of only 20 min without any sample pre-treatment. The features of the biosensor suggest its potential application for real time, fast and sensitive screening of FN in water samples as an analytical tool for the monitoring of the water quality.This work received financial support from DIONISOS Project (Retos Colaboracion RTC-2017-6222-5). The ICN2 is funded by the CERCA programme/Generalitat de Catalunya. The ICN2 is supported by the Severo Ochoa Centres of Excellence programme, funded by the Spanish Research Agency (AEI, grant no. SEV-2017-0706)Ramirez-Priego, P.; Estévez, M.; Díaz-Luisravelo, HJ.; Manclus Ciscar, JJ.; Montoya, Á.; Lechuga, LM. (2021). Real-time monitoring of fenitrothion in water samples using a silicon nanophotonic biosensor. Analytica Chimica Acta. 1152:1-9. https://doi.org/10.1016/j.aca.2021.338276S191152Sánchez-Santed, F., Colomina, M. T., & Herrero Hernández, E. (2016). Organophosphate pesticide exposure and neurodegeneration. Cortex, 74, 417-426. doi:10.1016/j.cortex.2015.10.003Chough, S. H., Mulchandani, A., Mulchandani, P., Chen, W., Wang, J., & Rogers, K. R. (2002). Organophosphorus Hydrolase-Based Amperometric Sensor: Modulation of Sensitivity and Substrate Selectivity. Electroanalysis, 14(4), 273-276. doi:10.1002/1521-4109(200202)14:43.0.co;2-5Richardson, J. R., Fitsanakis, V., Westerink, R. H. S., & Kanthasamy, A. G. (2019). Neurotoxicity of pesticides. Acta Neuropathologica, 138(3), 343-362. doi:10.1007/s00401-019-02033-9Giordano, G., Afsharinejad, Z., Guizzetti, M., Vitalone, A., Kavanagh, T. J., & Costa, L. G. (2007). Organophosphorus insecticides chlorpyrifos and diazinon and oxidative stress in neuronal cells in a genetic model of glutathione deficiency. Toxicology and Applied Pharmacology, 219(2-3), 181-189. doi:10.1016/j.taap.2006.09.016Çakir, Ş., & Sarikaya, R. (2005). Genotoxicity testing of some organophosphate insecticides in the Drosophila wing spot test. Food and Chemical Toxicology, 43(3), 443-450. doi:10.1016/j.fct.2004.11.010Rahman, M. F., Mahboob, M., Danadevi, K., Saleha Banu, B., & Grover, P. (2002). Assessment of genotoxic effects of chloropyriphos and acephate by the comet assay in mice leucocytes. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 516(1-2), 139-147. doi:10.1016/s1383-5718(02)00033-5Yeh, S.-P., Sung, T.-G., Chang, C.-C., Cheng, W., & Kuo, C.-M. (2005). Effects of an organophosphorus insecticide, trichlorfon, on hematological parameters of the giant freshwater prawn, Macrobrachium rosenbergii (de Man). Aquaculture, 243(1-4), 383-392. doi:10.1016/j.aquaculture.2004.10.017Smith, A. G., & Gangolli, S. D. (2002). Organochlorine chemicals in seafood: occurrence and health concerns. Food and Chemical Toxicology, 40(6), 767-779. doi:10.1016/s0278-6915(02)00046-7Kumar, P., Kim, K.-H., & Deep, A. (2015). Recent advancements in sensing techniques based on functional materials for organophosphate pesticides. 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M., & George, K. M. (2010). Mass Spectrometric Analyses of Organophosphate Insecticide Oxon Protein Adducts. Environmental Health Perspectives, 118(1), 11-19. doi:10.1289/ehp.0900824Wang, J., Chatrathi, M. P., Mulchandani, A., & Chen, W. (2001). Capillary Electrophoresis Microchips for Separation and Detection of Organophosphate Nerve Agents. Analytical Chemistry, 73(8), 1804-1808. doi:10.1021/ac001424eWatanabe, E., Kanzaki, Y., Tokumoto, H., Hoshino, R., Kubo, H., & Nakazawa, H. (2001). Enzyme-Linked Immunosorbent Assay Based on a Polyclonal Antibody for the Detection of the Insecticide Fenitrothion. Evaluation of Antiserum and Application to the Analysis of Water Samples. Journal of Agricultural and Food Chemistry, 50(1), 53-58. doi:10.1021/jf0108359Hua, X., Yang, J., Wang, L., Fang, Q., Zhang, G., & Liu, F. (2012). Development of an Enzyme Linked Immunosorbent Assay and an Immunochromatographic Assay for Detection of Organophosphorus Pesticides in Different Agricultural Products. PLoS ONE, 7(12), e53099. doi:10.1371/journal.pone.0053099Liu, G., & Lin, Y. (2005). Electrochemical Sensor for Organophosphate Pesticides and Nerve Agents Using Zirconia Nanoparticles as Selective Sorbents. Analytical Chemistry, 77(18), 5894-5901. doi:10.1021/ac050791tMane, P. C., Shinde, M. D., Varma, S., Chaudhari, B. P., Fatehmulla, A., Shahabuddin, M., … Chaudhari, R. D. (2020). Highly sensitive label-free bio-interfacial colorimetric sensor based on silk fibroin-gold nanocomposite for facile detection of chlorpyrifos pesticide. Scientific Reports, 10(1). doi:10.1038/s41598-020-61130-yEnsafi, A. A., Rezaloo, F., & Rezaei, B. (2017). Electrochemical Determination of Fenitrothion Organophosphorus Pesticide Using Polyzincon Modified-glassy Carbon Electrode. Electroanalysis, 29(12), 2839-2846. doi:10.1002/elan.201700406Qi, P., Wang, J., Wang, X., Wang, X., Wang, Z., Xu, H., … Wang, X. (2018). Sensitive determination of fenitrothion in water samples based on an electrochemical sensor layered reduced graphene oxide, molybdenum sulfide (MoS2)-Au and zirconia films. Electrochimica Acta, 292, 667-675. doi:10.1016/j.electacta.2018.09.187Kant, R. (2019). Surface plasmon resonance based fiber–optic nanosensor for the pesticide fenitrothion utilizing Ta2O5 nanostructures sequestered onto a reduced graphene oxide matrix. Microchimica Acta, 187(1). doi:10.1007/s00604-019-4002-8Zinoviev, K. E., Gonzalez-Guerrero, A. B., Dominguez, C., & Lechuga, L. M. (2011). Integrated Bimodal Waveguide Interferometric Biosensor for Label-Free Analysis. Journal of Lightwave Technology, 29(13), 1926-1930. doi:10.1109/jlt.2011.2150734Fernández Gavela, A., Grajales García, D., Ramirez, J., & Lechuga, L. (2016). Last Advances in Silicon-Based Optical Biosensors. Sensors, 16(3), 285. doi:10.3390/s16030285Maldonado, J., Estévez, M.-C., Fernández-Gavela, A., González-López, J. J., González-Guerrero, A. B., & Lechuga, L. M. (2020). Label-free detection of nosocomial bacteria using a nanophotonic interferometric biosensor. The Analyst, 145(2), 497-506. doi:10.1039/c9an01485cHuertas, C. S., Fariña, D., & Lechuga, L. M. (2016). Direct and Label-Free Quantification of Micro-RNA-181a at Attomolar Level in Complex Media Using a Nanophotonic Biosensor. ACS Sensors, 1(6), 748-756. doi:10.1021/acssensors.6b00162Maldonado, J., González-Guerrero, A. B., Domínguez, C., & Lechuga, L. M. (2016). Label-free bimodal waveguide immunosensor for rapid diagnosis of bacterial infections in cirrhotic patients. Biosensors and Bioelectronics, 85, 310-316. doi:10.1016/j.bios.2016.04.095González-Guerrero, A. B., Maldonado, J., Dante, S., Grajales, D., & Lechuga, L. M. (2016). Direct and label-free detection of the human growth hormone in urine by an ultrasensitive bimodal waveguide biosensor. 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Olive Leaves as Biotemplates for Enhanced Solar-Light Harvesting by a Titania-Based Solid

Olive leaves (by-product from olive oil production in olive mills) were used as biotemplates to synthesize a titania-based artificial olive leaf (AOL). Scanning electron microscopy (SEM) images of AOL showed the successful replication of trichomes and internal structure channels present in olive leaves. The BET surface area of AOL was 52 m2·g−1. X-ray diffraction (XRD) and Raman spectra revealed that the resulting solid was in the predominantly-anatase crystalline form (7.5 nm average particle size). Moreover, the synthesis led to a red-shift in light absorption as compared to reference anatase (gap energies of 2.98 and 3.2 eV, respectively). The presence of surface defects (as evidenced by X-ray photoelectron spectroscopy, XPS, and electron paramagnetic resonance spectroscopy, EPR) and doping elements (e.g., 1% nitrogen, observed by elemental analysis and XPS) could account for that. AOL was preliminarily tested as a catalyst for hydrogen production through glycerol photoreforming and exhibited an activity 64% higher than reference material Evonik P25 under solar irradiation and 144% greater under ultraviolet radiation, (under voltage) UV

Label-free detection of nosocomial bacteria using a nanophotonic interferometric biosensor

Nosocomial infections are a major concern at the worldwide level. Early and accurate identification of nosocomial pathogens is crucial to provide timely and adequate treatment. A prompt response also prevents the progression of the infection to life-threatening conditions, such as septicemia or generalized bloodstream infection. We have implemented two highly sensitive methodologies using an ultrasensitive photonic biosensor based on a bimodal waveguide interferometer (BiMW) for the fast detection of Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA), two of the most prevalent bacteria associated with nosocomial infections. For that, we have developed a biofunctionalization strategy based on the use of a PEGylated silane (silane-PEG-COOH) which provides a highly resistant and bacteria-repelling surface, which is crucial to specifically detect each bacterium. Two different biosensor assays have been set under standard buffer conditions: One based on a specific direct immunoassay employing polyclonal antibodies for the detection of P. aeruginosa and another one employing aptamers for the direct detection of MRSA. The biosensor immunoassay for P. aeruginosa is fast (it only takes 12 min) and specific and has experimentally detected concentrations down to 800 cfu mL (cfu: Colony forming unit). The second one relies on the use of an aptamer that specifically detects penicillin-binding protein 2a (PBP2a), a protein only expressed in the MRSA mutant, providing a photonic biosensor with the ability to identify the resistant pathogen MRSA and differentiate it from methicillin-susceptible S. aureus (MSSA). Direct, label-free, and selective detection of whole MRSA bacteria has been achieved, making possible the direct detection of also 800 cfu mL. According to the signal-to-noise (S/N) ratio of the device, a theoretical limit of detection (LOD) of around 49 and 29 cfu mL was estimated for P. aeruginosa and MRSA, respectively. Both results obtained under standard conditions reveal the great potential this interferometric biosensor device has as a versatile and specific tool for bacterial detection and quantification, providing a rapid method for the identification of nosocomial pathogens within the clinical requirements of sensitivity for the diagnosis of infections

Detection and Quantification of HspX Antigen in Sputum Samples Using Plasmonic Biosensing : Toward a Real Point-of-Care (POC) for Tuberculosis Diagnosis

Advancements that occurred during the last years in the diagnosis of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis infection, have prompted increased survival rates of patients. However, limitations related to the inefficiency of an early detection still remain; some techniques and laboratory methods do not have enough specificity and most instruments are expensive and require handling by trained staff. In order to contribute to a prompt and effective diagnosis of tuberculosis, we report the development of a portable, user-friendly, and low-cost biosensor device for its early detection. By using a label-free surface plasmon resonance (SPR) biosensor, we have established a direct immunoassay for the direct detection and quantification of the heat shock protein X (HspX) of Mtb, a well-established biomarker of this pathogen, directly in pretreated sputum samples. The method relies on highly specific monoclonal antibodies that are previously immobilized on the plasmonic sensor surface. This technology allows for the direct detection of the biomarker without amplification steps, showing a limit of detection (LOD) of 0.63 ng mL-1 and a limit of quantification (LOQ) of 2.12 ng mL-1. The direct analysis in pretreated sputum shows significant differences in the HspX concentration in patients with tuberculosis (with concentration levels in the order of 116-175 ng mL-1) compared with non-tuberculosis infected patients (values below the LOQ of the assay)

Trends and challenges of refractometric nanoplasmonic biosensors : a review

Motivated by potential benefits such as sensor miniaturization, multiplexing opportunities and higher sensitivities, refractometric nanoplasmonic biosensing has profiled itself in a short time span as an interesting alternative to conventional Surface Plasmon Resonance (SPR) biosensors. This latter conventional sensing concept has been subjected during the last decades to strong commercialization, thereby strongly leaning on well-developed thin-film surface chemistry protocols. Not surprisingly, the examples found in literature based on this sensing concept are generally characterized by extensive analytical studies of relevant clinical and diagnostic problems. In contrast, the more novel Localized Surface Plasmon Resonance (LSPR) alternative finds itself in a much earlier, and especially, more fundamental stage of development. Driven by new fabrication methodologies to create nanostructured substrates, published work typically focuses on the novelty of the presented material, its optical properties and its use - generally limited to a proof-of-concept - as a label-free biosensing scheme. Given the different stages of development both SPR and LSPR sensors find themselves in, it becomes apparent that providing a comparative analysis of both concepts is not a trivial task. Nevertheless, in this review we make an effort to provide an overview that illustrates the progress booked in both fields during the last five years. First, we discuss the most relevant advances in SPR biosensing, including interesting analytical applications, together with different strategies that assure improvements in performance, throughput and/or integration. Subsequently, the remaining part of this work focuses on the use of nanoplasmonic sensors for real label-free biosensing applications. First, we discuss the motivation that serves as a driving force behind this research topic, together with a brief summary that comprises the main fabrication methodologies used in this field. Next, the sensing performance of LSPR sensors is examined by analyzing different parameters that can be invoked in order to quantitatively assess their overall sensing performance. Two aspects are highlighted that turn out to be especially important when trying to maximize their sensing performance, being (1) the targeted functionalization of the electromagnetic hotspots of the nanostructures, and (2) overcoming inherent negative influence that stem from the presence of a high refractive index substrate that supports the nanostructures. Next, although few in numbers, an overview is given of the most exhaustive and diagnostically relevant LSPR sensing assays that have been recently reported in literature, followed by examples that exploit inherent LSPR characteristics in order to create highly integrated and high-throughput optical biosensors. Finally, we discuss a series of considerations that, in our opinion, should be addressed in order to bring the realization of a stand-alone LSPR biosensor with competitive levels of sensitivity, robustness and integration (when compared to a conventional SPR sensor) much closer to reality

Design of a surface plasmon resonance immunoassay for therapeutic drug monitoring of amikacin

The therapeutic drug monitoring (TDM) of pharmaceutical drugs with narrow therapeutic ranges is of great importance in the clinical setting. It provides useful information towards the enhancement of drug therapies, aiding in dosage control and toxicity risk management. Amikacin is an aminoglycoside antibiotic commonly used in neonatal therapies that is indicated for TDM due to the toxicity risks inherent in its use. Current techniques for TDM such as high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) are costly, time consuming, and cannot be performed at the site of action. Over the last decades, surface plasmon resonance (SPR) biosensors have become increasingly popular in clinical diagnostics due to their ability to detect biomolecular interactions in real-time. We present an SPR-based competitive immunoassay for the detection of the antibiotic amikacin, suitable for TDM in both adults and neonates. We have obtained high specificity and sensitivity levels with an IC value of 1.4 ng/mL and a limit of detection of 0.13 ng/mL, which comfortably comply with the drug's therapeutic range. Simple dilution of serum can therefore be sufficient to analyze low-volume real samples from neonates, increasing the potential of the methodology for TDM. Compared to current TDM conventional methods, this SPR-based immunoassay can provide advantages such as simplicity, potential portability, and label-free measurements with the possibility of high throughput. This work is the foundation towards the development of an integrated, simple use, highly sensitive, fast, and point-of-care sensing platform for the opportune TDM of antibiotics and other drugs in a clinical setting