Combination of RT-LAMP and fluorescence spectroscopy using chemometric techniques for an ultra-sensitive and rapid alternative for the detection of SARS-CoV-2.

The increased spread of COVID-19 caused by SARS-CoV-2 has made it necessary to develop more efficient, fast, accurate, specific, sensitive and easy-to-use detection platforms to overcome the disadvantages of gold standard methods (RT-qPCR). Here an approach was developed for the detection of the SARS-CoV-2 virus using the loop-mediated isothermal amplification (LAMP) technique for SARS-CoV-2 RNA target amplification in samples of nasopharyngeal swabs. The discrimination between positive and negative SARS-CoV-2 samples was achieved by using fluorescence spectra generated by the excitation of the LAMP's DNA intercalator dye at λ497 nm in a fluorescence spectrophotometer and chemometric tools. Exploratory analysis of the 83 sample spectra using principal component analysis (PCA) indicated a trend in differentiation between positive and negative samples resulting from the peak emission of the fluorescent dye. The classification was performed by partial least squares discriminant analysis (PLS-DA) achieving a sensitivity, a specificity and an accuracy of 100%, 95% and 89%, respectively for the discrimination between negative and positive samples from 1.58 to 0.25 ng L-1 after LAMP amplification. Therefore, this study indicates that the use of the LAMP technique in fluorescence spectroscopy may offer a fast (<1 hour), sensitive and low-cost method.

Development and Application of an SPR Nanobiosensor Based on AuNPs for the Detection of SARS-CoV-2 on Food Surfaces.

A new transmission route of SARS-CoV-2 through food was recently considered by the World Health Organization (WHO), and, given the pandemic scenario, the search for fast, sensitive, and low-cost methods is necessary. Biosensors have become a viable alternative for large-scale testing because they overcome the limitations of standard techniques. Herein, we investigated the ability of gold spherical nanoparticles (AuNPs) functionalized with oligonucleotides to detect SARS-CoV-2 and demonstrated their potential to be used as plasmonic nanobiosensors. The loop-mediated isothermal amplification (LAMP) technique was used to amplify the viral genetic material from the raw virus-containing solution without any preparation. The detection of virus presence or absence was performed by ultraviolet-visible (UV-Vis) absorption spectroscopy, by monitoring the absorption band of the surface plasmonic resonance (SPR) of the AuNPs. The displacement of the peak by 525 nm from the functionalized AuNPs indicated the absence of the virus (particular region of gold). On the other hand, the region ~300 nm indicated the presence of the virus when RNA bound to the functionalized AuNPs. The nanobiosensor system was designed to detect a region of the N gene in a dynamic concentration range from 0.1 to 50 × 103 ng·mL-1 with a limit of detection (LOD) of 1 ng·mL-1 (2.7 × 103 copy per µL), indicating excellent sensitivity. The nanobiosensor was applied to detect the SARS-CoV-2 virus on the surfaces of vegetables and showed 100% accuracy compared to the standard quantitative reverse transcription polymerase chain reaction (RT-qPCR) technique. Therefore, the nanobiosensor is sensitive, selective, and simple, providing a viable alternative for the rapid detection of SARS-CoV-2 in ready-to-eat vegetables.

A systematic review of the advancement on colorimetric nanobiosensors for SARS-CoV-2 detection.

The current pandemic of the acute severe respiratory syndrome coronavirus 2 (SARS-CoV-2) killed about 6.4 million and infected more than 600 million individuals by august of 2022, and researchers worldwide are searching for fast and selective approaches for this virus detection. Colorimetric biosensors are an excellent alternative because they are sensitive, simple, fast, and low-cost for rapid detection of SARS-CoV-2 compared to standard Enzyme-linked immunosorbent assay (ELISA) and Polymerase Chain Reaction (PCR) techniques. This study systematically searched and reviewed literature data related to colorimetric biosensors in detecting SARS-CoV-2 viruses, recovered from the Scopus (n = 16), Web of Science (n = 19), PubMed (n = 19), and Science Direct (n = 17) databases totalizing n = 71 articles. Data were analyzed for the type of nanomaterial, biorecognition material at the detection limit (LOD), and devices designed for diagnostics. The most applied nanomaterial were gold nanoparticles, in their original form and hybrid in quantum dots and core-shell. In addition, we show high specificity in point-of-care (POC) diagnostic devices as a faster and cheaper alternative for clinical diagnosis. Finally, the highlights of the colorimetric biosensor developed for diagnostic devices applied in swabs, surgical masks, and lateral flow immunoassays were presented.

A systematic review of the advancement on colorimetric nanobiosensors for SARS-CoV-2 detection

The current pandemic of the acute severe respiratory syndrome coronavirus 2 (SARS-CoV-2) killed about 6.4 million and infected more than 600 million individuals by august of 2022, and researchers worldwide are searching for fast and selective approaches for this virus detection. Colorimetric biosensors are an excellent alternative because they are sensitive, simple, fast, and low-cost for rapid detection of SARS-CoV-2 compared to standard Enzyme-linked immunosorbent assay (ELISA) and Polymerase Chain Reaction (PCR) techniques. This study systematically searched and reviewed literature data related to colorimetric biosensors in detecting SARS-CoV-2 viruses, recovered from the Scopus (n=16), Web of Science (n=19), PubMed (n=19), and Science Direct (n=17) databases totalizing n=71 articles. Data were analyzed for the type of nanomaterial, biorecognition material at the detection limit (LOD), and devices designed for diagnostics. The most applied nanomaterial were gold nanoparticles, in their original form and hybrid in quantum dots and core-shell. In addition, we show high specificity in point-of-care (POC) diagnostic devices as a faster and cheaper alternative for clinical diagnosis. Finally, the highlights of the colorimetric biosensor developed for diagnostic devices applied in swabs, surgical masks, and lateral flow immunoassays were presented. Graphical

ATR-FTIR spectroscopy and chemometrics as a quick and simple alternative for discrimination of SARS-CoV-2 infected food of animal origin.

Alternative routes such as virus transmission or cross-contamination by food have been suggested, due to reported cases of SARS-CoV-2 in frozen chicken wings and fish or seafood. Delay in routine testing due to the dependence on the PCR technique as the standard method leads to greater virus dissemination. Therefore, alternative detection methods such as FTIR spectroscopy emerge as an option. Here, we demonstrate a fast (3 min), simple and reagent-free methodology using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy for discrimination of food (chicken, beef and fish) contaminated with the SARS-CoV-2 virus. From the IR spectra of the samples, the "bio-fingerprint" (800 - 1900 cm-1) was selected to investigate the distinctions caused by the virus contamination. Exploratory analysis of the spectra, using Principal Component of Analysis (PCA), indicated the differentiation in the data due to the presence of single bands, marked as contamination from nucleic acids including viral RNA. Furthermore, the partial least squares discriminant analysis (PLS-DA) classification model allowed for discrimination of each matrix in its pure form and its contaminated counterpart with sensitivity, specificity and accuracy of 100 %. Therefore, this study indicates that the use of ATR-FTIR can offer a fast and low cost and not require chemical reagents and with minimal sample preparation to detect the SARS-CoV-2 virus in food matrices, ensuring food safety and non-dissemination by consumers.

Updating the use of nano-biosensors as promising devices for the diagnosis of coronavirus family members: A systematic review.

Coronavidae viruses, such as SARS-CoV, SARS-CoV-2, and MERS-CoV, cause severe lower respiratory tract infection, acute respiratory distress syndrome and extrapulmonary manifestations, such as diarrhea and fever, eventually leading to death. Fast, accurate, reproductible, and cost-effective SARS-CoV-2 identification can be achieved employing nano-biosensors, reinforcing conventional methodologies to avoid the spread of COVID-19 within and across communities. Nano-biosensors built using gold, silver, graphene, In2O3 nanowire and iron oxide nanoparticles, Quantum Dots and carbon nanofibers have been successfully employed to detect specific virus antigens - nucleic acid sequences and/or proteins -or host antibodies produced in response to viral infection. Biorecognition counterpart molecules have been immobilized on the surface of these nanomaterials, leading to selective virus detection by optical or electrochemical transducer systems. This systematic review assessed studies on described and tested immunonsensors and genosensors designed from distinct nanomaterials available at the Pubmed, Scopus, and Science Direct databases. Twenty-three nano biosensors were found suitable for unequivocal coronavirus detection in clinical samples. Nano-biosensors coupled to RT-LAMP/RT-PCR assays can optimize RNA extraction, reduce analysis times and/or eliminate sophisticated instrumentation. Although promising for the diagnosis of Coronavidae family members, further trials in large populations must be adequately and rigorously conducted to address nano-biosensor applicability in the clinical practice for early coronavirus infection detection.