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High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections

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A special issue of Biomedicines (ISSN 2227-9059). This special issue belongs to the section "Molecular and Translational Medicine".

Deadline for manuscript submissions: closed (28 February 2023) | Viewed by 20273

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Dr. Hideya Kawasaki

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Institute for NanoSuit Research, Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan
Interests: pathology; virus; neuroscience; electron microscopy; nanoSuit
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Dear Colleagues,

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other infections are a serious threat to public health and the global economy. The World Health Organization (WHO) designated the coronavirus disease 2019 (COVID-19) outbreak as a pandemic in March 2020. According to the WHO, the global cumulative number of novel coronavirus infections exceeds 300 million to date. The rapid identification and isolation of patients diagnosed with SARS-CoV-2 is important for preventing transmission.

Point of care testing (POCT) is applicable to a variety of areas of medicine and can make a significant difference in patient care. Lateral flow assays (LFAs) are critical in POCT. An LFA is simple-to-use diagnostic equipment that is used to confirm the presence or absence of a target analyte, such as SARS-CoV-2 or other infective agents, or biomarkers in humans or animals, or pollutants in drinking water, food, or animal feed. However, its clinical utility has been questioned due to its limited sensitivity. Numerous strategies are used to improve sensitivity and quantitative detection, e.g., employing several visualization methods, using different labeling reporters, integrating LFAs with other detection methods, resulting in their benefiting from both LFAs and the advantages of integrated detection devices for SARS-CoV-2 or other infections. This Special Issue invites submissions of novel and innovative original studies as well as comprehensive reviews on this topic.

Dr. Hideya Kawasaki
Guest Editor

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Keywords

lateral flow assay
point of care testing
SARS-CoV-2
high sensitivity
detection device
labeling reporters
antigens
pathogens

Related Special Issue

High-Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections Volume II in Biomedicines (1 article)

Published Papers (11 papers)

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9 pages, 897 KiB

Open AccessArticle

Clinical Performance of Lateral Flow Assay for Cryptosporidium spp. Diagnosis

by Miriam Campos-Ruiz, Clara Flamarich, Anabel Fernández-Navarro, Silvia Roure, Laura Martin, Pablo Pillado, Pere-Joan Cardona and Gema Fernández-Rivas

Biomedicines 2023, 11(8), 2140; https://doi.org/10.3390/biomedicines11082140 - 29 Jul 2023

Viewed by 643

Abstract

Cryptosporidium spp. is an apicomplexan protozoan parasite associated with gastroenteritis in humans. In 2018, Spain showed 1511 confirmed cases, with a growing trend since 2014. Despite this fact, Cryptosporidium spp. is not usually routinely examined when a parasitological study is ordered, although accurate diagnosis is fundamental to prevent the spread of the illness. The main objectives of the present work is to demonstrate the circulation and to study the epidemiology of cryptosporidiosis in patients who were being tested for the presence of Cryptosporidium spp. parasites in the faeces in the Metropolitan North Area of Barcelona, Maresme, and Vallés Occidental using a two-step algorithm. The stool samples were analysed using the Cryptosporidium/Giardia spp. immunochromatographic test; the positive samples were visualised under a microscope using auramine staining. The proportion of Cryptosporidium spp. cases was around 2% in the studied patients, with a pronounced seasonal incidence peak in late summer–early autumn. In our cohort, weight loss was the main symptom related to confirmed cases. The mean age of confirmed patients was 19 years old, and they were younger than the unconfirmed group. Cryptosporidium spp. is one of the parasites that currently circulate in many areas in Europe. Prevalence must be taken into account for active searching. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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10 pages, 511 KiB

Open AccessArticle

Evaluation of Four Rapid Antigen Tests for the Detection of SARS-CoV-2 Infection with Nasopharyngeal Swabs

by Ho-Jae Lim, Min-Young Park, Young-Hyun Baek, Hyeon-Seo Lee, Inhee Kim, Youngjin Kwon, Youngshin You, Kyoungwoo Nam, Jae-Hyun Yang, Min-Jin Kim, Nae Yu, Yong-Hak Sohn, Jung-Eun Park and Yong-Jin Yang

Biomedicines 2023, 11(3), 701; https://doi.org/10.3390/biomedicines11030701 - 24 Feb 2023

Cited by 2 | Viewed by 1345

Abstract

Owing to the high transmissibility of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, the capacity of testing systems based on the gold standard real-time reverse transcription–polymerase chain reaction (rRT-PCR) is limited. Rapid antigen tests (RATs) can substantially contribute to the prevention of community transmission, but their further assessment is required. Here, using 1503 nasopharyngeal swabs, we compared the diagnostic performance of four RAT kits (Abbott Panbio™ COVID-19 Ag Rapid Test, SD Biosensor Standard™ Q COVID-19 Ag Test, Humasis COVID-19 Ag Test, and SG Medical Acrosis COVID-19 Ag Test) to the cycle threshold (Ct) values obtained from rRT-PCR. The precision values, area under the curve values, SARS-CoV-2 variant detection ability, and non-SARS-CoV-2 specificity of all four kits were similar. An assay using the Acrosis kit had a significantly better positive detection rate with a higher recall value and cut-off value than that using the other three RAT kits. During the current COVID-19 pandemic, the Acrosis kit is an effective tool to prevent the spread of SARS-CoV-2 in communities. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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11 pages, 509 KiB

Open AccessArticle

Clinical Validation of GenBody COVID-19 Ag, Nasal and Nasopharyngeal Rapid Antigen Tests for Detection of SARS-CoV-2 in European Adult Population

by Karolina Wegrzynska, Jaroslaw Walory, Radoslaw Charkiewicz, Marzena Anna Lewandowska, Izabela Wasko, Aleksandra Kozinska, Piotr Majewski and Anna Baraniak

Biomedicines 2023, 11(2), 493; https://doi.org/10.3390/biomedicines11020493 - 08 Feb 2023

Cited by 5 | Viewed by 1803

Abstract

Accurate and rapid identification of COVID-19 is critical for effective patient treatment and disease outcomes, as well as the prevention of SARS-CoV-2 transmission. Rapid antigen tests (RATs) for identifying SARS-CoV-2 are simpler, faster and less expensive than molecular assays. Any new product to be considered a medical device is subject to evaluation and data analysis to verify the in vitro diagnostic ability to achieve its intended purpose. Clinical validation of such a test is a prerequisite before clinical application. This study was a clinical validation on adult Europeans of GenBody COVID-19 Ag, nasal and nasopharyngeal RATs. A set of 103 positive and 301 negative from nose and nasopharynx samples confirmed by RT-qPCR were examined. The tests were safe to use and showed 100% specificity in both specimens, and high sensitivity of 94.17% (95%CI 87.75% to 97.83%) and 97.09% (95%CI 91.72% to 99.4%), respectively. The parameters were significantly better for samples with higher virus loads (the highest for CT ≤ 25). The GenBody COVID-19 Ag RATs are inexpensive (compared to RT-qPCR), reliable and rapid with high sensitivity and specificity, making them suitable for diagnosis and timely isolation and treatment of COVID-19 patients, contributing to the better control of virus spread. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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10 pages, 785 KiB

Open AccessArticle

Prospective Clinical Evaluation of the Diagnostic Accuracy of a Highly Sensitive Rapid Antigen Test Using Silver Amplification Technology for Emerging SARS-CoV-2 Variants

by Kazuaki Obata, Kei Miyakawa, Toshiki Takei, Atsuhiko Wada, Yasuyoshi Hatayama, Hideaki Kato, Yayoi Kimura, Hisakuni Sekino, Junichi Katada and Akihide Ryo

Biomedicines 2022, 10(11), 2801; https://doi.org/10.3390/biomedicines10112801 - 03 Nov 2022

Cited by 1 | Viewed by 1632

Abstract

The COVID-19 pandemic caused by SARS-CoV-2 remains a serious health concern worldwide due to outbreaks of SARS-CoV-2 variants that can escape vaccine-acquired immunity and infect and transmit more efficiently. Therefore, an appropriate testing method for COVID-19 is essential for effective infection control and the prevention of local outbreaks. Compared to reverse-transcription polymerase chain reaction (RT-PCR) tests, antigen tests are used for simple point-of-care testing, enabling the identification of viral infections. In this study, we tested the clinical usefulness of the FUJIFILM COVID-19 Ag test, an antigen test based on silver amplification and immunochromatographic technology. The FUJIFILM COVID-19 Ag test was shown to detect a lower viral concentration as compared to other conventional kits without significant performance loss in detecting prevalent SARS-CoV-2 variants. We tested nasopharyngeal and nasal swabs from a single patient during two different epidemic periods dominated by various SARS-CoV-2 variants. We observed that the sensitivity of the FUJIFILM COVID-19 Ag test was 95.7% and 85.7% in nasopharyngeal and nasal swabs, respectively. These results suggest that the FUJIFILM COVID-19 Ag test is highly sensitive and applicable when RT-PCR testing is unavailable. Furthermore, these results indicate that high-frequency testing using nasal swab specimens may be a valuable screening strategy. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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14 pages, 3350 KiB

Open AccessArticle

Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay

by Aleksandra Anna Zasada, Ewa Mosiej, Marta Prygiel, Maciej Polak, Karol Wdowiak, Kamila Formińska, Robert Ziółkowski, Kamil Żukowski, Kasper Marchlewicz, Adam Nowiński, Julia Nowińska, Waldemar Rastawicki and Elżbieta Malinowska

Biomedicines 2022, 10(9), 2329; https://doi.org/10.3390/biomedicines10092329 - 19 Sep 2022

Cited by 13 | Viewed by 2041

Abstract

Rapid and accurate detection and identification of pathogens in clinical samples is essential for all infection diseases. However, in the case of epidemics, it plays a key role not only in the implementation of effective therapy but also in limiting the spread of the epidemic. In this study, we present the application of two nucleic acid isothermal amplification methods—reverse transcription helicase dependent amplification (RT-HDA) and reverse transcription loop-mediated amplification (RT-LAMP)—combined with lateral flow assay as the tools for the rapid detection of SARS-CoV-2, the etiological agent of COVID-19, which caused the ongoing global pandemic. In order to optimize the RT-had, the LOD was 3 genome copies per reaction for amplification conducted for 10–20 min, whereas for RT-LAMP, the LOD was 30–300 genome copies per reaction for a reaction conducted for 40 min. No false-positive results were detected for RT-HDA conducted for 10 to 90 min, but false-positive results occurred when RT-LAMP was conducted for longer than 40 min. We concluded that RT-HDA combined with LFA is more sensitive than RT-LAMP, and it is a good alternative for the development of point-of-care tests for SARS-CoV-2 detection as this method is simple, inexpensive, practical, and does not require qualified personnel to perform the test and interpret its results. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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9 pages, 711 KiB

Open AccessArticle

Clinical Performance of Self-Collected Nasal Swabs and Antigen Rapid Tests for SARS-CoV-2 Detection in Resource-Poor Settings

by Nádia Sitoe, Júlia Sambo, Nédio Mabunda, Neuza Nguenha, Jorfélia Chilaúle, Júlio Rafael, Anésio Macicame, Imelda Chelene, Chishamiso Mudenyanga, Jillian Sacks, Sofia Viegas, Osvaldo Loquiha and Ilesh Jani

Biomedicines 2022, 10(9), 2327; https://doi.org/10.3390/biomedicines10092327 - 19 Sep 2022

Cited by 2 | Viewed by 1749

Abstract

Background: In resource-poor countries, antigen-based rapid tests (Ag-RDTs) performed at primary healthcare and community settings improved access to SARS-CoV-2 diagnostics. However, the technical skills and biosafety requirements inherent to nasopharyngeal and oropharyngeal (OP) specimens limit the scale-up of SARS-CoV-2 testing. The collection of nasal-swabs is programmatically viable, but its performance has not been evaluated in resource-poor settings. Methods: We first evaluated the performance of SteriPack self-collected nasal swabs for the detection of SARS-CoV-2 by real-time PCR in 1498 consecutively enrolled patients with suspected infection. Next, we evaluated the clinical performance of three nasal swab-based Ag-RDTs against real-time PCR on OP specimens. Results: The sensitivity of nasal swabs was 80.6% [95% CI: 75.3–85.2%] compared to OP specimens. There was a good correlation (r = 0.58; p < 0.0001) between Ct values of 213 positive cases obtained using nasal and OP swabs. Our findings show sensitivities of 79.7% [95% CI: 73.3–85.1%] for Panbio COVID-19 Ag-RDT, 59.6% [95% CI: 55.2–63.8%] for COVIOS Ag-RDT, and 78.0% [95% CI: 73.5–82.0%] for the LumiraDx SARS-CoV-2 Ag-RDT. Conclusions: In our setting, the COVIOS Ag-RDT did not meet WHO requirements. Nasal swab-based Ag-RDTs for SARS-CoV-2 detection constitute a viable and accurate diagnostic option in resource-poor settings. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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12 pages, 3340 KiB

Open AccessArticle

Development of a Fully Automated Desktop Analyzer and Ultrahigh Sensitivity Digital Immunoassay for SARS-CoV-2 Nucleocapsid Antigen Detection

by Ryotaro Chiba, Kei Miyakawa, Kotaro Aoki, Takamitsu J. Morikawa, Yoshiki Moriizumi, Takuma Degawa, Yoshiyuki Arai, Osamu Segawa, Kengo Tanaka, Hideji Tajima, Susumu Arai, Hisatoshi Yoshinaga, Ryohei Tsukada, Akira Tani, Haruhito Fuji, Akinobu Sato, Yoshikazu Ishii, Kazuhiro Tateda, Akihide Ryo and Toru Yoshimura

Biomedicines 2022, 10(9), 2291; https://doi.org/10.3390/biomedicines10092291 - 15 Sep 2022

Cited by 4 | Viewed by 2485

Abstract

Background: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak has had a significant impact on public health and the global economy. Several diagnostic tools are available for the detection of infectious diseases, with reverse transcription-polymerase chain reaction (RT-PCR) testing specifically recommended for viral RNA detection. However, this diagnostic method is costly, complex, and time-consuming. Although it does not have sufficient sensitivity, antigen detection by an immunoassay is an inexpensive and simpler alternative to RT-PCR. Here, we developed an ultrahigh sensitivity digital immunoassay (d-IA) for detecting SARS-CoV-2 nucleocapsid (N) protein as antigens using a fully automated desktop analyzer based on a digital enzyme-linked immunosorbent assay. Methods: We developed a fully automated d-IA desktop analyzer and measured the viral N protein as an antigen in nasopharyngeal (NP) swabs from patients with coronavirus disease. We studied nasopharyngeal swabs of 159 and 88 patients who were RT-PCR-negative and RT-PCR-positive, respectively. Results: The limit of detection of SARS-CoV-2 d-IA was 0.0043 pg/mL of N protein. The cutoff value was 0.029 pg/mL, with a negative RT-PCR distribution. The sensitivity of RT-PCR-positive specimens was estimated to be 94.3% (83/88). The assay time was 28 min. Conclusions: Our d-IA system, which includes a novel fully automated desktop analyzer, enabled detection of the SARS-CoV-2 N-protein with a comparable sensitivity to RT-PCR within 30 min. Thus, d-IA shows potential for SARS-CoV-2 detection across multiple diagnostic centers including small clinics, hospitals, airport quarantines, and clinical laboratories. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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20 pages, 2629 KiB

Open AccessArticle

A Protein Microarray-Based Respiratory Viral Antigen Testing Platform for COVID-19 Surveillance

by Sungjun Beck, Rie Nakajima, Algis Jasinskas, Timothy J. Abram, Sun Jin Kim, Nader Bigdeli, Delia F. Tifrea, Jenny Hernandez-Davies, D. Huw Davies, Per Niklas Hedde, Philip L. Felgner and Weian Zhao

Biomedicines 2022, 10(9), 2238; https://doi.org/10.3390/biomedicines10092238 - 09 Sep 2022

Cited by 4 | Viewed by 2509

Abstract

High-throughput and rapid screening testing is highly desirable to effectively combat the rapidly evolving COVID-19 pandemic co-presents with influenza and seasonal common cold epidemics. Here, we present a general workflow for iterative development and validation of an antibody-based microarray assay for the detection of a respiratory viral panel: (a) antibody screening to quickly identify optimal reagents and assay conditions, (b) immunofluorescence assay design including signal amplification for low viral titers, (c) assay characterization with recombinant proteins, inactivated viral samples and clinical samples, and (d) multiplexing to detect a panel of common respiratory viruses. Using RT-PCR-confirmed SARS-CoV-2 positive and negative pharyngeal swab samples, we demonstrated that the antibody microarray assay exhibited a clinical sensitivity and specificity of 77.2% and 100%, respectively, which are comparable to existing FDA-authorized antigen tests. Moreover, the microarray assay is correlated with RT-PCR cycle threshold (Ct) values and is particularly effective in identifying high viral titers. The multiplexed assay can selectively detect SARS-CoV-2 and influenza virus, which can be used to discriminate these viral infections that share similar symptoms. Such protein microarray technology is amenable for scale-up and automation and can be broadly applied as a both diagnostic and research tool. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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8 pages, 379 KiB

Open AccessArticle

High Diagnostic Accuracy of a Novel Lateral Flow Assay for the Point-of-Care Detection of SARS-CoV-2

by Irene Giberti, Elisabetta Costa, Alexander Domnich, Valentina Ricucci, Vanessa De Pace, Giada Garzillo, Giulia Guarona and Giancarlo Icardi

Biomedicines 2022, 10(7), 1558; https://doi.org/10.3390/biomedicines10071558 - 30 Jun 2022

Cited by 3 | Viewed by 2274

Abstract

Highly accurate lateral flow immunochromatographic tests (LFTs) are an important public health tool to tackle the ongoing COVID-19 pandemic. The aim of this study was to assess the comparative diagnostic performance of the novel ND COVID-19 LFT under real-world conditions. A total of 400 nasopharyngeal swab specimens with a wide range of viral loads were tested in both reverse-transcription polymerase chain reaction and ND LFT. The overall sensitivity and specificity were 85% (95% CI: 76.7–90.7%) and 100% (95% CI: 98.7–100%), respectively. There was a clear association between the false-negative rate and sample viral load: the sensitivity parameters for specimens with cycle threshold values of <25 (>3.95 × 10⁶ copies/mL) and ≥30 (≤1.29 × 10⁵ copies/mL) were 100% and 50%, respectively. The performance was maximized in testing samples with viral loads ≥1.29 × 10⁵ copies/mL. These findings suggest that the ND LFT is sufficiently accurate and useful for mass population screening programs, especially in high-prevalence and resource-constrained settings or during periods when the epidemic curve is rising. Other public health implications were also discussed. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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12 pages, 1818 KiB

Open AccessArticle

Rapid and Low-Cost Detection and Quantification of SARS-CoV-2 Antibody Titers of ICU Patients with Respiratory Deterioration Using a Handheld Thermo-Photonic Device

by Damber Thapa, Nakisa Samadi, Andrew Baker, Claudia dos Santos, Uriel Trahtemberg and Nima Tabatabaei

Biomedicines 2022, 10(6), 1424; https://doi.org/10.3390/biomedicines10061424 - 15 Jun 2022

Cited by 2 | Viewed by 1679

Abstract

While research suggests that COVID-19 vaccines are effective in producing anti-SARS-CoV-2 antibodies that reduce the risk of COVID-19 and its potentially severe complications, how long these antibodies persist after the infection/vaccination is unknown. Longitudinal studies and rapid and scalable platforms are needed for large-scale sero-diagnosis and vaccine evaluation. In this study, we examine the efficacy of our recently-developed handheld thermo-photonic device for rapid and low-cost assessment of the adaptive immune response of COVID⁺ and COVID⁻ patients admitted to the intensive care unit (ICU) at a local hospital due to respiratory deterioration. Antibody testing included detection and quantification of IgG and IgM via thermo-photonic sensing of a commercially available COVID-19 IgG/IgM rapid test as well as standard measurements with quantitative enzyme-linked immunosorbent assays (qELISA). The results demonstrate that the thermo-photonic reader in conjunction with COVID-19 IgG/IgM test cassettes can detect and quantify IgG levels in COVID-19 antibody assays within the clinically relevant range and with a high correlation to those obtained from qELISA. We also found that the IgG antibody is more reliable for detecting individuals with an adaptive immune response to SARS-CoV-2 compared to the IgM antibody. The developed reader offers a low-cost, portable, and scalable solution for accessing the antibody titer of individuals against SARS-CoV-2 and can be used in local hospital settings. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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7 pages, 1158 KiB

Open AccessBrief Report

Evaluation of a Commercial Point-of-Care RT-LAMP Assay for Rapid Detection of SARS-CoV-2

by Janet Hei Yin Law, Wai Sing Chan, Tsun Leung Chan, Edmond Shiu Kwan Ma and Bone Siu Fai Tang

Biomedicines 2023, 11(9), 2344; https://doi.org/10.3390/biomedicines11092344 - 23 Aug 2023

Viewed by 673

Abstract

The goal of this study was to evaluate the performance of a commercial reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay (Detect COVID-19 Test) in the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A total of 202 human respiratory and viral culture specimens were tested retrospectively. The performance of the Detect COVID-19 Test was comparable to that of commercial real-time polymerase chain reaction assays (sensitivity: 93.42%; specificity: 100%), and better than that of the rapid antigen test (sensitivity: 48.00%; specificity: 100%) for specimens with threshold cycle (Ct) values of less than 30. The Beta, Delta, and Omicron variants of concern were successfully detected. With their simplicity of use and good assay sensitivity, point-of-care RT-LAMP assays may be a viable option for SARS-CoV-2 testing at home, or in regions without sophisticated laboratory facilities. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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