The Lateral Flow Immunoassay (LFIA) is by far one of the most successful analytical platforms to perform the on-site detection of target substances. LFIA can be considered as a sort of lab-in-a-hand ...and, together with other point-of-need tests, has represented a paradigm shift from sample-to-lab to lab-to-sample aiming to improve decision making and turnaround time. The features of LFIAs made them a very attractive tool in clinical diagnostic where they can improve patient care by enabling more prompt diagnosis and treatment decisions. The rapidity, simplicity, relative cost-effectiveness, and the possibility to be used by nonskilled personnel contributed to the wide acceptance of LFIAs. As a consequence, from the detection of molecules, organisms, and (bio)markers for clinical purposes, the LFIA application has been rapidly extended to other fields, including food and feed safety, veterinary medicine, environmental control, and many others. This review aims to provide readers with a 10-years overview of applications, outlining the trends for the main application fields and the relative compounded annual growth rates. Moreover, future perspectives and challenges are discussed.
We have developed a simple and accurate biosensor based on a chemiluminescent (CL)-lateral flow immunoassay (LFIA) method integrated in a smartphone to quantitatively detect salivary cortisol. The ...biosensor is based on a direct competitive immunoassay using peroxidase–cortisol conjugate, detected by adding the chemiluminescent substrate luminol/enhancer/hydrogen peroxide. The smartphone camera is used as light detector, for image acquisition and data handling via a specific application. We 3D-printed simple accessories to adapt the smartphone. The system comprises a cartridge, which houses the LFIA strip, and a smartphone adaptor with a plano-convex lens and a cartridge-insertion slot. This provides a mini-darkbox and aligned optical interface between the camera and the LFIA membrane for acquiring CL signals. The method is simple and fast, with a detection limit of 0.3ng/mL. It provides quantitative analysis in the range of 0.3–60ng/mL, which is adequate for detecting salivary cortisol in the clinically accepted range. It could thus find application in the growing area of home-self-diagnostic device technology for clinical biomarker monitoring, overcoming the current difficulties in achieving sensitive and quantitative information with conventional systems taking the advantage of smartphone connectivity and the enhanced performance of the included camera.
•A biosensor based on chemiluminescent lateral flow immunoassay was developed.•Smartphone's camera was used as a detector for chemiluminescent signals.•Smartphone's accessories were realized for acquiring chemiluminescent signals.•The biosensor allowed to quantify cortisol in saliva sample.
Simultaneous measurement of different substances from a single sample is an emerging issue for achieving efficient and high-throughput detection in several fields of application. Although ...immunoanalytical techniques have well-established and prevailing advantages over alternative screening analytical platforms, one of the incoming challenges for immunoassay is exact multiplexing. Lateral flow immunoassay (LFIA) is a leading immunoanalytical technique for onsite analysis, thanks to its simplicity, rapidity, and cost-effectiveness. Moreover, LFIA architecture is adaptable to multiplexing, and is therefore a possible answer to the pressing demand of multiplexing point-of-need analysis. This review presents an overview of diverse approaches for multiplex LFIA, with a special focus on strategies based on new types of magnetic, fluorescent, and colored labels.
A rapid test for detecting total immunoglobulins directed towards the nucleocapsid protein (N) of severe acute syndrome coronavirus 2 (SARS CoV-2) was developed, based on a multi-target lateral flow ...immunoassay comprising two test lines. Both test lines bound to several classes of immunoglobulins (G, M, and A). Specific anti-SARS immunoglobulins were revealed by a colorimetric probe formed by N and gold nanoparticles. Targeting the total antibodies response to infection enabled achieving 100% diagnostic specificity (95.75–100, C.I. 95%, n = 85 healthy and with other infections individuals) and 94.6% sensitivity (84.9–98.9, C.I. 95%, n = 62 SARS CoV-2 infected subjects) as early as 7 days post confirmation of positivity. Agreeing results with a reference serological ELISA were achieved, except for the earlier detection capability of the rapid test. Follow up of the three seroconverting patients endorsed the hypothesis of the random rise of the different immunoglobulins and strengthened the ‘total antibodies’ approach for the trustworthy detection of serological response to SARS CoV-2 infection.
Display omitted A rapid test based on the lateral flow immunoassay technology was established to detect the total serological response to the SARS CoV-2 infection.
•A sensitive and specific lateral flow immunoassay for detecting antibodies to Covid-19.•Broad-specific agents to capture total immunoglobulins enabled increasing the sensitivity.•Two test lines were combined to guarantee no false positive results.•Diagnosis based on the multi-target LFIA can complement molecular assays.
Natural toxin (for example mycotoxin and phycotoxin) contamination of food is of safety and economic concern, so much effort is devoted to the development of screening methods which enable the toxins ...to be continuously and widely monitored in food and feed. More generally speaking, rapid and non-instrumental assays for detection of a variety of food contaminants are generating ever-increasing scientific and technological interest because they enable high-throughput, economical, on-site monitoring of such contaminants. Among rapid methods for first-level screening of food contaminants, lateral-flow immunoassay (LFIA), also named immunochromatographic assay or immune-gold colloid immunoassay, has recently attracted scientific and industrial interest because of its attractive property of enabling very rapid, one-step, in-situ analysis. This review focuses on new aspects of the development and optimization of lateral-flow devices for mycotoxin and phycotoxin detection, including strategies for management of matrix interference and, particularly, for investigation of the improvements achieved by signal-enhancing strategies or by application of non-gold nanoparticle signal reporters.
Figure 1
Competitive lateral flow immunoassay for myco- or phycotoxin: the
Test
zone is formed by adsorbing a conjugate of the target compound (
toxin
);
Control
zone is formed by anti-species antibodies (
white
), reporters are specific (anti-toxin antibodies,
black
) and non-specific (
grey
) antibodies labelled with gold nanoparticles (
GNP
). Focalization of GNP-labelled antibodies determines a visible/detectable colour appearance on both the Test and Control lines, which can be related to analyte amount in a liquid sample.
In the current paradigm for molecular imprinting, the imprinted binding sites exist as a consequence of the polymerization process around templates, and the properties of nonimprinted polymers (NIPs) ...have largely been overlooked. Thus, nothing can be affirmed a priori concerning the binding properties of NIPs. We propose an alternative view where the imprinting effect is due to the presence of a template molecule that enhances the pre-existing binding properties of a polymer. If a NIP shows no binding properties toward a target molecule, the corresponding imprinted polymer (MIP) will show a weak imprinting effect. On the other hand, if a NIP shows binding properties toward a target molecule, the corresponding MIP will show a significant imprinting effect. To verify this hypothesis, we prepared a 96-member combinatorial polymeric library in the absence of any template molecule. This library was screened for several potential ligands, and with no exceptions, the composition of the best-binding NIP produced a MIP with excellent binding properties, whereas a low-binding NIP formulation produced a MIP with comparable low binding. To validate these results, the binding properties toward naproxen and ibuprofen were measured for two combinatorial libraries of polymers prepared in the presence (MIP library) and the absence (NIP library) of the template molecule. The experiment’s results showed a correlation between the apparent affinity constants measured for the NIP and MIP libraries, confirming the proposed hypothesis. Moreover, for closely related molecules, it was shown that binding selectivity is an emergent property derived from the imprinting process and not a property of NIPs.
Paper-based lateral-flow immunoassays (LFIAs) have achieved considerable commercial success and their impact in diagnostics is continuously growing. LFIA results are often obtained by visualizing by ...the naked eye color changes in given areas, providing a qualitative information about the presence/absence of the target analyte in the sample. However, this platform has the potential to provide ultrasensitive quantitative analysis for several applications. Indeed, LFIA is based on well-established immunological techniques, which have known in the last year great advances due to the combination of highly sensitive tracers, innovative signal amplification strategies and last-generation instrumental detectors. All these available progresses can be applied also to the LFIA platform by adapting them to a portable and miniaturized format. This possibility opens countless strategies for definitively turning the LFIA technique into an ultrasensitive quantitative method. Among the different proposals for achieving this goal, the use of enzyme-based immunoassay is very well known and widespread for routine analysis and it can represent a valid approach for improving LFIA performances. Several examples have been recently reported in literature exploiting enzymes properties and features for obtaining significative advances in this field. In this review, we aim to provide a critical overview of the recent progresses in highly sensitive LFIA detection technologies, involving the exploitation of enzyme-based amplification strategies. The features and applications of the technologies, along with future developments and challenges, are also discussed.
Mycotoxins are toxic metabolites of molds which can contaminate food and beverages. Because of their acute and chronic toxicity, they can have harmful effects when ingested or inhaled, posing severe ...risks to human health. Contemporary analytical methods have the sensitivity required for contamination detection and quantification, but the direct application of these methods on real samples is not straightforward because of matrix complexity, and clean-up and preconcentration steps are needed, more and more requiring the application of highly selective solid-phase extraction materials. Molecularly imprinted polymers (MIPs) are artificial receptors mimicking the natural antibodies that are increasingly being used as a solid phase in extraction methods where selectivity towards target analytes is mandatory. In this review, the state-of-the-art about molecularly imprinted polymers as solid-phase extraction materials in mycotoxin contamination analysis will be discussed, with particular attention paid to the use of mimic molecules in the synthesis of mycotoxin-imprinted materials, to the application of these materials to food real samples, and to the development of advanced extraction methods involving molecular imprinting technology.
Abstract
Anti-immigration rhetoric in the mass media has intensified over the last two decades, potentially decreasing prosocial behavior and increasing outgroup hostility toward immigrants, and ...fostering ingroup favoritism toward natives. We aim to understand the effects of negative and positive discourses about immigration on prosociality at different levels of societal ethnic diversity. In two studies (student sample, nationally representative sample), we conduct a survey and a 3X3 between-subject experiment, including money-incentivized behavioral games measuring prosociality. We manipulate media representations of immigrants and the probability of interacting with immigrants (the latter measuring diversity). Results show that negative news affects prosociality as a function of the probability of interacting with immigrants. Negative portrayals increase altruism and trustworthiness in ethnically homogenous settings relative to unknown and ethnically-mixed contexts. These results are stronger for right-wing and high-prejudice respondents. Moreover, negative media portrayals of immigrants increase the testosterone-cortisol ratio, which is a proxy for proneness to social aggression. Negative news also increases outgroup-related perceived health risk, outgroup anxiety and outgroup threat less in ethnically-homogeneous contexts. Overall, negative portrayals of immigrants generate physiological and emotional hostility toward the outgroup, and ingroup favoritism in economic transactions, possibly determining efficiency losses in ethnically-diverse markets, relative to ethnically-homogeneous markets.
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► The development of a high sensitive lateral flow immunoassay is described. ► The developed assay allowed aflatoxin M1 detection in milk at level required by EU regulations. ► ...Article describes advances in lateral flow technology towards high sensitivity. ► A simple and rapid sample pre-treatment was proposed to overthrow matrix interference.
A high sensitive immunoassay-based lateral flow device for semi-quantitatively determine aflatoxin M1 (AFM1) in milk was developed. Investigation and optimization of the competitor design and of the gold-labelling strategy allowed the attainment of the ultra-sensitive assessment of AFM1 contamination at nanograms per litre level (LOD 20ngL−1, IC50 99ngL−1), as requested by European regulations. A one order of magnitude detectability enhancement in comparison to previously reported gold colloid immunochromatographic assays for this toxin was obtained.
Direct detection of the target toxin in milk could be obtained by acquiring images of the strips and correlating intensities of the coloured lines with analyte concentrations. The one-step assay can be completed in 17min, including a very simple and rapid sample preparation, which allowed the application of the assay to milk samples which differ in fat and protein contents. Although imprecise (mean RSD about 30%), the method proved to be accurate and sensitive enough to allow the correct attribution of sample as compliant or non-compliant according to EU legislation in force. Agreeing results to those of a reference ELISA were obtained on 40 milk samples by matrix-matched calibration in pasteurized milk.
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