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Lateral Flow Immunoassay: Advances and Applications

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A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Biosensors".

Deadline for manuscript submissions: closed (20 October 2021) | Viewed by 48445

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Dr. Fabio Di Nardo

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Università degli Studi di Torino | UNITO · Dipartimento di Chimica, Turin, Italy
Interests: bioanalytical chemistry; immunoassays development and validation; lateral flow immunoassay, immunochromatography, paper-based biosensors; noble metal nanoparticles synthesis and characterization; food safety; mycotoxins; Point-of-Care Test and Point-of-Need Test

Special Issue Information

Dear Colleagues,

Worldwide, there is an undeniable increasing demand for reliable rapid tests for point-of-need applications (i.e., all the analyses performed directly where the sample is obtained). Immunochemical methods are one of the most successful and versatile strategies for point-of-need applications. Among the plethora of immunoassay-based tests, the lateral flow immunoassay (LFIA) embodies the ideal technique to be used in a decentralized testing strategy, thanks to its simplicity, rapidity, cost-effectiveness, and lack of requirement of equipment or technical expertise. LFIA is very useful especially in low-resource field environments and in developing countries that cannot afford standard analytical instrumentation to perform analyses. Moreover, LFIA can be the right ally of standard analytical methods in developed countries, ensuring high sample throughput and rapid decision making. Historically, point-of-need tests have been considered as less accurate than those used in centralized testing environments. However, there has been a terrific evolution in LFIA technologies in the past decades, and LFIAs are becoming well suited to replace laboratory-based immunoassays in decentralized testing locations. Moreover, LFIA can be the leading technique in meeting the revised ASSURED criteria, i.e., the REASSURED criteria (real-time connectivity, ease of specimen collection, affordable, sensitive, specific, user-friendly, rapid and robust, equipment-free or simple and environmentally friendly, and deliverable to end-users).

This Special Issue on “Lateral Flow Immunoassay: Advances and Applications” aims to highlight recent advances in the technique, covering the use of new labels and different detection systems, the use of different molecular recognition elements and new strategies to improve analytical performances (e.g., sensitivity, accuracy, reproducibility, shelf-life, multiplexing), etc. Moreover, new relevant applications of traditional LFIA concerning but not limited to food and feed safety and clinical, veterinary, and environmental control can be highlighted. Both original research papers and review articles describing the current state-of-the-art in lateral flow immunoassay are welcome.

Dr. Fabio Di Nardo
Guest Editor

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Keywords

Lateral flow immunoassay
Lateral flow assay
Gold nanoparticles
Noble metal nanoparticles
Antibodies
Paper-based biosensors
Point-of-care testing
Point-of-need testing
ASSURED criteria
REASSURED criteria

Published Papers (9 papers)

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Research

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15 pages, 2903 KiB

Open AccessArticle

Development of a Dengue Virus Serotype-Specific Non-Structural Protein 1 Capture Immunochromatography Method

by Kanaporn Poltep, Emi E. Nakayama, Tadahiro Sasaki, Takeshi Kurosu, Yoshiki Takashima, Juthamas Phadungsombat, Nathamon Kosoltanapiwat, Borimas Hanboonkunupakarn, Sarin Suwanpakdee, Hisham A. Imad, Narinee Srimark, Chiaki Kitamura, Atsushi Yamanaka, Akio Okubo, Tatsuo Shioda and Pornsawan Leaungwutiwong

Sensors 2021, 21(23), 7809; https://doi.org/10.3390/s21237809 - 24 Nov 2021

Cited by 2 | Viewed by 2969

Abstract

Four serotypes of dengue virus (DENV), type 1 to 4 (DENV-1 to DENV-4), exhibit approximately 25–40% of the difference in the encoded amino acid residues of viral proteins. Reverse transcription of RNA extracted from specimens followed by PCR amplification is the current standard method of DENV serotype determination. However, since this method is time-consuming, rapid detection systems are desirable. We established several mouse monoclonal antibodies directed against DENV non-structural protein 1 and integrated them into rapid DENV detection systems. We successfully developed serotype-specific immunochromatography systems for all four DENV serotypes. Each system can detect 10⁴ copies/mL in 15 min using laboratory and clinical isolates of DENV. No cross-reaction between DENV serotypes was observed in these DENV isolates. We also confirmed that there was no cross-reaction with chikungunya, Japanese encephalitis, Sindbis, and Zika viruses. Evaluation of these systems using serum from DENV-infected individuals indicated a serotype specificity of almost 100%. These assay systems could accelerate both DENV infection diagnosis and epidemiologic studies in DENV-endemic areas. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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18 pages, 5776 KiB

Open AccessFeature PaperArticle

Reducing Unspecific Protein Adsorption in Microfluidic Papers Using Fiber-Attached Polymer Hydrogels

by Alexander Ritter von Stockert, Anna Luongo, Markus Langhans, Thomas Brandstetter, Jürgen Rühe, Tobias Meckel and Markus Biesalski

Sensors 2021, 21(19), 6348; https://doi.org/10.3390/s21196348 - 23 Sep 2021

Cited by 5 | Viewed by 2312

Abstract

Microfluidic paper combines pump-free water transport at low cost with a high degree of sustainability, as well as good availability of the paper-forming cellulosic material, thus making it an attractive candidate for point-of-care (POC) analytics and diagnostics. Although a number of interesting demonstrators for such paper devices have been reported to date, a number of challenges still exist, which limit a successful transfer into marketable applications. A strong limitation in this respect is the (unspecific) adsorption of protein analytes to the paper fibers during the lateral flow assay. This interaction may significantly reduce the amount of analyte that reaches the detection zone of the microfluidic paper-based analytical device (µPAD), thereby reducing its overall sensitivity. Here, we introduce a novel approach on reducing the nonspecific adsorption of proteins to lab-made paper sheets for the use in µPADs. To this, cotton linter fibers in lab-formed additive-free paper sheets are modified with a surrounding thin hydrogel layer generated from photo-crosslinked, benzophenone functionalized copolymers based on poly-(oligo-ethylene glycol methacrylate) (POEGMA) and poly-dimethyl acrylamide (PDMAA). This, as we show in tests similar to lateral flow assays, significantly reduces unspecific binding of model proteins. Furthermore, by evaporating the transport fluid during the microfluidic run at the end of the paper strip through local heating, model proteins can almost quantitatively be accumulated in that zone. The possibility of complete, almost quantitative protein transport in a µPAD opens up new opportunities to significantly improve the signal-to-noise (S/N) ratio of paper-based lateral flow assays. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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12 pages, 2804 KiB

Open AccessArticle

Silver-Assembled Silica Nanoparticles in Lateral Flow Immunoassay for Visual Inspection of Prostate-Specific Antigen

by Hyung-Mo Kim, Jaehi Kim, Sungje Bock, Jaehyun An, Yun-Sik Choi, Xuan-Hung Pham, Myeong Geun Cha, Bomi Seong, Wooyeon Kim, Yoon-Hee Kim, Hobeom Song, Jung-Won Kim, Seung-min Park, Sang Hun Lee, Won-Yeop Rho, Sangchul Lee, Dae Hong Jeong, Ho-Young Lee and Bong-Hyun Jun

Sensors 2021, 21(12), 4099; https://doi.org/10.3390/s21124099 - 15 Jun 2021

Cited by 15 | Viewed by 3516

Abstract

Prostate-specific antigen (PSA) is the best-known biomarker for early diagnosis of prostate cancer. For prostate cancer in particular, the threshold level of PSA <4.0 ng/mL in clinical samples is an important indicator. Quick and easy visual detection of the PSA level greatly helps in early detection and treatment of prostate cancer and reducing mortality. In this study, we developed optimized silica-coated silver-assembled silica nanoparticles (SiO₂@Ag@SiO₂ NPs) that were applied to a visual lateral flow immunoassay (LFIA) platform for PSA detection. During synthesis, the ratio of silica NPs to silver nitrate changed, and as the synthesized NPs exhibited distinct UV spectra and colors, most optimized SiO₂@Ag@SiO₂ NPs showed the potential for early prostate cancer diagnosis. The PSA detection limit of our LFIA platform was 1.1 ng/mL. By applying each SiO₂@Ag@SiO₂ NP to the visual LFIA platform, optimized SiO₂@Ag@SiO₂ NPs were selected in the test strip, and clinical samples from prostate cancer patients were successfully detected as the boundaries of non-specific binding were clearly seen and the level of PSA was <4 ng/mL, thus providing an avenue for quick prostate cancer diagnosis and early treatment. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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17 pages, 2791 KiB

Open AccessArticle

From Smartphone Lateral Flow Immunoassay Screening to Direct MS Analysis: Development and Validation of a Semi-Quantitative Direct Analysis in Real-Time Mass Spectrometric (DART-MS) Approach to the Analysis of Deoxynivalenol

by Ariadni Geballa-Koukoula, Arjen Gerssen and Michel W. F. Nielen

Sensors 2021, 21(5), 1861; https://doi.org/10.3390/s21051861 - 07 Mar 2021

Cited by 9 | Viewed by 3092

Abstract

In current food safety monitoring, lateral flow immunoassays (LFIAs) are widely used for rapid food contaminant screening. Recent advances include smartphone readouts, offering semi-quantitative analysis of LFIAs with time, location, and data transfer in case of on-site testing. Following the screening, the next step in the EU regulations is confirmation by, e.g., liquid chromatography-tandem mass spectrometry (LC-MS/MS). In this work, using direct analysis in real time ambient ionization and triple quadrupole MS/MS (DART-QqQ-MS/MS), we achieved rapid confirmation of the identity of the substance(s) causing the LFIA result. In the workflow proposed, an individual performs the (on-site) smartphone LFIA screening, and when the result is suspect, an identification LFIA (ID-LFIA) strip is developed with the same sample extract. The ID-LFIA can be dissociated and rapidly analyzed in a control laboratory with DART-QqQ-MS/MS. The ID-LFIA consists of multiple lines of monoclonal antibodies against the mycotoxin deoxynivalenol, acting as a bioaffinity trap. The ID-LFIA/DART-QqQ-MS/MS approach has been developed and validated, along with the screening smartphone LFIA, and has demonstrated its applicability by analyzing incurred and spiked samples. The developed approach has been critically compared with our previous direct electrospray ionization MS method and was found to provide highly complementary information on the total deoxynivalenol contamination in the sample. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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16 pages, 5367 KiB

Open AccessArticle

Paper-Based Competitive Immunochromatography Coupled with an Enzyme-Modified Electrode to Enable the Wireless Monitoring and Electrochemical Sensing of Cotinine in Urine

by Nutcha Larpant, Pramod K. Kalambate, Tautgirdas Ruzgas and Wanida Laiwattanapaisal

Sensors 2021, 21(5), 1659; https://doi.org/10.3390/s21051659 - 28 Feb 2021

Cited by 14 | Viewed by 3377

Abstract

This paper proposes a combined strategy of using paper-based competitive immunochromatography and a near field communication (NFC) tag for wireless cotinine determination. The glucose oxidase labeled cotinine antibody specifically binds free cotinine in a sample, whereas the unoccupied antibody attached to BSA-cotinine at the test line on a lateral flow strip. The glucose oxidase on the strip and an assistant pad in the presence of glucose generated H₂O₂ and imposed the Ag oxidation on the modified electrode. This enabled monitoring of immunoreaction by either electrochemical measurement or wireless detection. Wireless sensing was realized for cotinine in the range of 100–1000 ng/mL (R² = 0.96) in PBS medium. Undiluted urine samples from non-smokers exhibited an Ag-oxidation rate three times higher than the smoker’s urine samples. For 1:8 diluted urine samples (smokers), the proposed paper-based competitive immunochromatography coupled with an enzyme-modified electrode differentiated positive and negative samples and exhibited cotinine discrimination at levels higher than 12 ng/mL. This novel sensing platform can potentially be combined with a smartphone as a reader unit. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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17 pages, 3607 KiB

Open AccessFeature PaperArticle

Double-Antigen Lateral Flow Immunoassay for the Detection of Anti-HIV-1 and -2 Antibodies Using Upconverting Nanoparticle Reporters

by Iida Martiskainen, Etvi Juntunen, Teppo Salminen, Karoliina Vuorenpää, Sherif Bayoumy, Tytti Vuorinen, Navin Khanna, Kim Pettersson, Gaurav Batra and Sheikh M. Talha

Sensors 2021, 21(2), 330; https://doi.org/10.3390/s21020330 - 06 Jan 2021

Cited by 17 | Viewed by 4334

Abstract

Rapid diagnostic tests (RDTs) are often used for the detection of anti-human immunodeficiency virus (HIV) antibodies in remote locations in low- and middle-income countries (LMIC) with low or limited access to central laboratories. The typical format of an RDT is a lateral flow assay (LFA) with visual interpretation prone to subjectivity. This risk of misinterpretation can be overcome with luminescent upconverting nanoparticle reporters (UCNPs) measured with a miniaturized easy-to-use reader instrument. An LFA with UCNPs for anti-HIV-1/2 antibodies was developed and the assay performance was evaluated extensively with challenging patient sample panels. Sensitivity (n = 145) of the UCNP-LFA was 96.6% (95% CI: 92.1–98.8%) and specificity (n = 309) was 98.7% (95% CI: 96.7–99.7%). Another set of samples (n = 200) was used for a comparison between the UCNP-LFA and a conventional visual RDT. In this comparison, the sensitivities for HIV-1 were 96.4% (95% CI: 89.8–99.3%) and 97.6% (95% CI: 91.6–99.7%), for the UCNP-LFA and conventional RDT, respectively. The specificity was 100% (95% CI: 96.4–100%) for both assays. The developed UCNP-LFA demonstrates the applicability of UCNPs for the detection of anti-HIV antibodies. The signal measurement is done by a reader instrument, which may facilitate automated result interpretation, archiving and transfer of data from de-centralized locations. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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14 pages, 2685 KiB

Open AccessArticle

Switching from Multiplex to Multimodal Colorimetric Lateral Flow Immunosensor

by Simone Cavalera, Fabio Di Nardo, Luca Forte, Francesca Marinoni, Matteo Chiarello, Claudio Baggiani and Laura Anfossi

Sensors 2020, 20(22), 6609; https://doi.org/10.3390/s20226609 - 18 Nov 2020

Cited by 11 | Viewed by 3183

Abstract

Multiplex lateral flow immunoassay (LFIA) is largely used for point-of-care testing to detect different pathogens or biomarkers in a single device. The increasing demand for multitargeting diagnostics requires multi-informative single tests. In this study, we demonstrated three strategies to upgrade standard multiplex LFIA to multimodal capacity. As a proof-of-concept, we applied the strategies to the differential diagnosis of Human Immunodeficiency Virus (HIV) infection, a widespread pathogen, for which conventional multiplex LFIA testing is well-established. In the new two-parameter LFIA (x²LFIA), we exploited color encoding, in which the binding of multiple targets occurs in one reactive band and the color of the probe reveals which one is present in the sample. By combining the sequential alignment of several reactive zones along the membrane of the LFIA strip and gold nanoparticles and gold nanostars for the differential visualization, in this demonstration, the x²LFIA can furnish information on HIV serotype and stage of infection in a single device. Three immunosensors were designed. The use of bioreagents as the capturing ligand anchored onto the membrane or as the detection ligand labelled with gold nanomaterials affected the performance of the x²LFIA. Higher detectability was achieved by the format involving the HIV-specific antigens as capturing agent and labelled secondary bioligands (anti-human immunoglobulins M and protein G) as the probes. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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Review

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33 pages, 2700 KiB

Open AccessReview

Ten Years of Lateral Flow Immunoassay Technique Applications: Trends, Challenges and Future Perspectives

by Fabio Di Nardo, Matteo Chiarello, Simone Cavalera, Claudio Baggiani and Laura Anfossi

Sensors 2021, 21(15), 5185; https://doi.org/10.3390/s21155185 - 30 Jul 2021

Cited by 189 | Viewed by 17292

Abstract

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. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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19 pages, 2901 KiB

Open AccessReview

Recent Advancements in Enzyme-Based Lateral Flow Immunoassays

by Donato Calabria, Maria Maddalena Calabretta, Martina Zangheri, Elisa Marchegiani, Ilaria Trozzi, Massimo Guardigli, Elisa Michelini, Fabio Di Nardo, Laura Anfossi, Claudio Baggiani and Mara Mirasoli

Sensors 2021, 21(10), 3358; https://doi.org/10.3390/s21103358 - 12 May 2021

Cited by 41 | Viewed by 6515

Abstract

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. Full article

(This article belongs to the Special Issue Lateral Flow Immunoassay: Advances and Applications)

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