Double lateral flow immunosensing of undeclared pork and chicken components of meat products.

Adulteration of meat products is a serious problem in the modern society. Consumption of falsified meat products can be hazardous to health and/or lead to violating religious dietary principles. To identify such products, rapid and simple test systems for point-of-need detection are in demand along with complex laboratory methods. This study presents the first double lateral flow (immunochromatographic) test system, which allows simultaneous revealing two prevalent types of falsifications-undeclared addition of pork and chicken components to meat products. In the proposed test system, porcine myoglobin (MG) and chicken immunoglobulin Y (IgY) were used as specific biomarkers recognizable by antibodies. Within the optimization of the analysis, the concentrations of the immune reagents and regimes of their application on the working membrane were selected, which provided minimal limits of detection (LODs) for both analytes. The developed test system enables the detection of MG and IgY with the LODs of 10 and 12 ng/mL, respectively, which accords to addition of 0.1% of the undeclared meat compounds. The applicability of the test system to control the composition of raw meat mixtures and cooked food products was confirmed. The developed approach can be considered as a promising tool for monitoring composition of meat products. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-024-05944-y.

The Development of Aptamer-Based Gold Nanoparticle Lateral Flow Test Strips for the Detection of SARS-CoV-2 S Proteins on the Surface of Cold-Chain Food Packaging.

The COVID-19 pandemic over recent years has shown a great need for the rapid, low-cost, and on-site detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, an aptamer-based colloidal gold nanoparticle lateral flow test strip was well developed to realize the visual detection of wild-type SARS-CoV-2 spike proteins (SPs) and multiple variants. Under the optimal reaction conditions, a low detection limit of SARS-CoV-2 S proteins of 0.68 nM was acquired, and the actual detection recovery was 83.3% to 108.8% for real-world samples. This suggests a potential tool for the prompt detection of SARS-CoV-2 with good sensitivity and accuracy, and a new method for the development of alternative antibody test strips for the detection of other viral targets.

Raman Scattering-Based Biosensing: New Prospects and Opportunities.

The growing interest in the development of new platforms for the application of Raman spectroscopy techniques in biosensor technologies is driven by the potential of these techniques in identifying chemical compounds, as well as structural and functional features of biomolecules. The effect of Raman scattering is a result of inelastic light scattering processes, which lead to the emission of scattered light with a different frequency associated with molecular vibrations of the identified molecule. Spontaneous Raman scattering is usually weak, resulting in complexities with the separation of weak inelastically scattered light and intense Rayleigh scattering. These limitations have led to the development of various techniques for enhancing Raman scattering, including resonance Raman spectroscopy (RRS) and nonlinear Raman spectroscopy (coherent anti-Stokes Raman spectroscopy and stimulated Raman spectroscopy). Furthermore, the discovery of the phenomenon of enhanced Raman scattering near metallic nanostructures gave impetus to the development of the surface-enhanced Raman spectroscopy (SERS) as well as its combination with resonance Raman spectroscopy and nonlinear Raman spectroscopic techniques. The combination of nonlinear and resonant optical effects with metal substrates or nanoparticles can be used to increase speed, spatial resolution, and signal amplification in Raman spectroscopy, making these techniques promising for the analysis and characterization of biological samples. This review provides the main provisions of the listed Raman techniques and the advantages and limitations present when applied to life sciences research. The recent advances in SERS and SERS-combined techniques are summarized, such as SERRS, SE-CARS, and SE-SRS for bioimaging and the biosensing of molecules, which form the basis for potential future applications of these techniques in biosensor technology. In addition, an overview is given of the main tools for success in the development of biosensors based on Raman spectroscopy techniques, which can be achieved by choosing one or a combination of the following approaches: (i) fabrication of a reproducible SERS substrate, (ii) synthesis of the SERS nanotag, and (iii) implementation of new platforms for on-site testing.

Hospitals and Laboratories on Paper-Based Sensors: A Mini Review.

With characters of low cost, portability, easy disposal, and high accuracy, as well as bulky reduced laboratory equipment, paper-based sensors are getting increasing attention for reliable indoor/outdoor onsite detection with nonexpert operation. They have become powerful analysis tools in trace detection with ultra-low detection limits and extremely high accuracy, resulting in their great popularity in medical detection, environmental inspection, and other applications. Herein, we summarize and generalize the recently reported paper-based sensors based on their application for mechanics, biomolecules, food safety, and environmental inspection. Based on the biological, physical, and chemical analytes-sensitive electrical or optical signals, extensive detections of a large number of factors such as humidity, pressure, nucleic acid, protein, sugar, biomarkers, metal ions, and organic/inorganic chemical substances have been reported via paper-based sensors. Challenges faced by the current paper-based sensors from the fundamental problems and practical applications are subsequently analyzed; thus, the future directions of paper-based sensors are specified for their rapid handheld testing.

A fluorescent nanosphere-based immunochromatography test strip for ultrasensitive and point-of-care detection of tetanus antibody in human serum.

Tetanus still possesses a high infection risk and leads a number of human deaths in poor nations. Point-of-care and ultrasensitive detection of tetanus antibody levels in serum is the key to decrease the risk of tetanus infection and improve the health of people. In this work, by using ultra bright fluorescent nanospheres (FNs) and portable lateral flow test strip (LFTS), a point-of-care and ultrasensitive sensing method has been developed for the detection of tetanus antibodies in human serum. This assay works quite well for tetanus antibodies in the concentration range from 0.0002 to 0.0220 IU/mL with a low detection limit of 0.00011 IU/mL, which is 100-fold lower than conventional gold-based LFTSs. The high sensitivity makes this method suitable for use to detect the low-abundant target in real samples. Besides, this cost-effective FN-based LFTS assay possesses good selectivity, high accuracy, and satisfactory reliability, which holds great potential as a robust candidate for routine medical diagnosis and rapid home testing. Graphical abstract.

How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

A simple approach was proposed to decrease the detection limit of sandwich lateral flow immunoassay (LFIA) by changing the conditions for binding between a polyvalent antigen and a conjugate of gold nanoparticles (GNPs) with antibodies. In this study, the potato virus Y (PVY) was used as the polyvalent antigen, which affects economically important plants in the Solanaceae family. The obtained polyclonal antibodies that are specific to PVY were characterized using a sandwich enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). For LFIA, the antibodies were conjugated with GNPs with a diameter of 17.4 ± 1.0 nm. We conducted LFIAs using GNP conjugates in a dried state on the test strip and after pre-incubation with a sample. Pre-incubating the GNP conjugates and sample for 30 s was found to decrease the detection limit by 60-fold from 330 ng∙mL-1 to 5.4 ng∙mL-1 in comparison with conventional LFIA. The developed method was successfully tested for its ability to detect PVY in infected and uninfected potato leaves. The quantitative results of the proposed LFIA with pre-incubation were confirmed by ELISA, and resulted in a correlation coefficient of 0.891. The proposed approach is rapid, simple, and preserves the main advantages of LFIA as a non-laboratory diagnostic method.

Simultaneous quantitation of cytokeratin-19 fragment and carcinoembryonic antigen in human serum via quantum dot-doped nanoparticles.

A novel quantum dot-doped polystyrene nanoparticles-based lateral flow test strips (QPs-LFTS) system was developed to simultaneously detect a cytokeratin-19 fragment (CYFRA 21-1) and carcinoembryonic antigen (CEA) in human serum to aid the diagnosis and prognosis of lung cancer. Quantum dot-doped carboxylate-functionalized polystyrene nanoparticles (QPs) were prepared and introduced as fluorescent reporters in QPs-LFTS. The detection was based on a sandwich immunoassay and performed on lateral flow test strips, with an assay time of 15min. The strips were read by a fluorescence strip reader to obtain the fluorescence peak heights of the test lines (HT) and the control line (HC). The ratio of HT/HC was used for quantitation. The QPs showed excellent photoproperties and good performance. Under optimal conditions, the QPs-LFTS system exhibited a wide linear range for CYFRA 21-1 (1.3-480ng/mL) and CEA (2.8-680ng/mL). The detection limits for CYFRA 21-1 and CEA were 0.16 and 0.35ng/mL, respectively. The recovery and reproducibility of the method were satisfactory. Furthermore, excellent correlations (n =120, R2 =0.9862, P<0.0001 for CYFRA 21-1; n =70, R2 =0.9509, P<0.0001 for CEA) were obtained between the QPs-LFTS and commercially available chemiluminescence immunoassay kits in clinical serum testing. The results indicate that this developed test system is highly efficient and is expected to be useful for early screening and prognosis evaluation for lung cancer patients.