Verification of lateral flow antigen tests for SARS-CoV-2 by qPCR directly from the test device

Czibere, Ludwig; Burggraf, Siegfried; Becker, Marc; Durner, Jürgen; Draenert, M.E.

doi:10.1016/j.dental.2022.03.005

Cited by 5 publications

(2 citation statements)

References 11 publications

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“…Based on this analysis, our 95% LoD threshold corresponds to 6.6 log10 RNA copies/mL. Studies on the analytical performance of RATs are diverse due to differences in study design and methods, sample size, and type of RAT evaluated, which demonstrated variable performance, and sometimes with reports of contradicting results for similar assays, making comparisons difficult [ 74 , 75 , 76 ]. Furthermore, variability in viral quantification, such as TCID 50 , plaque-forming units (pfu), or RT-PCR-based viral RNA copies, introduce bias to make comparisons [ 77 ].…”

Section: Resultsmentioning

confidence: 99%

Development and Analytical Evaluation of a Point-of-Care Electrochemical Biosensor for Rapid and Accurate SARS-CoV-2 Detection

Meshesha,

Sardar,

Supekar

et al. 2023

Sensors

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The COVID-19 pandemic has underscored the critical need for rapid and accurate screening and diagnostic methods for potential respiratory viruses. Existing COVID-19 diagnostic approaches face limitations either in terms of turnaround time or accuracy. In this study, we present an electrochemical biosensor that offers nearly instantaneous and precise SARS-CoV-2 detection, suitable for point-of-care and environmental monitoring applications. The biosensor employs a stapled hACE-2 N-terminal alpha helix peptide to functionalize an in situ grown polypyrrole conductive polymer on a nitrocellulose membrane backbone through a chemical process. We assessed the biosensor’s analytical performance using heat-inactivated omicron and delta variants of the SARS-CoV-2 virus in artificial saliva (AS) and nasal swab (NS) samples diluted in a strong ionic solution, as well as clinical specimens with known Ct values. Virus identification was achieved through electrochemical impedance spectroscopy (EIS) and frequency analyses. The assay demonstrated a limit of detection (LoD) of 40 TCID50/mL, with 95% sensitivity and 100% specificity. Notably, the biosensor exhibited no cross-reactivity when tested against the influenza virus. The entire testing process using the biosensor takes less than a minute. In summary, our biosensor exhibits promising potential in the battle against pandemic respiratory viruses, offering a platform for the development of rapid, compact, portable, and point-of-care devices capable of multiplexing various viruses. The biosensor has the capacity to significantly bolster our readiness and response to future viral outbreaks.

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Section: Resultsmentioning

confidence: 99%

Development and Analytical Evaluation of a Point-of-Care Electrochemical Biosensor for Rapid and Accurate SARS-CoV-2 Detection

Meshesha,

Sardar,

Supekar

et al. 2023

Sensors

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“…Rapid and robust detection plays a vital role in precise diagnosis and effective disease control. − Among these, the lateral flow immunoassay (LFIA) for rapid antigen detection can be effectively performed in the laboratory and in point-of-care testing (POCT) due to its advantages of rapidity, simplicity, accessibility, and readability. , However, the gold nanoparticle (GNP)-based conventional LFIA suffers from low sensitivity, false positives, and false negatives, which may be suitable for initial screening of the population. − To address these challenges, various signal enhancement strategies have been investigated to improve the sensing performance. − Emerging nanozymes attract great interest due to their high enzyme activities, good biocompatibility, and chemical stability, which have been widely used in biosensing. − Nanozymes feature the merits: (1) their own colors having colorimetric signals and (2) providing signal enhancement through catalytic activity, which have good prospects for possible signal tags in the LFIA. To date, peroxidase-like nanozymes have been widely used in the LFIA, − which may encounter issues such as longer color development time, weaker signal intensity, and inconvenient surface modifications compared to natural enzymes. − On the other hand, molecular recognition elements with high specificity and good affinity are another key factor in developing the LFIA.…”

Section: Introductionmentioning

confidence: 99%

Sensitive Lateral Flow Immunoassay Based on Specific Peptide and Superior Oxidase Mimics with a Universal Dual-Mode Significant Signal Amplification

et al. 2023

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Rapid and sensitive antigen detection using a lateral flow immunoassay (LFIA) is crucial for diagnosing infectious diseases due to its simplicity, speed, and user-friendly features. However, it remains a critical issue to explore specific biorecognition elements and powerful signal amplification. In this study, taking SARS-CoV-2 as a proof of concept, a specific peptide, WFLNDSELIML, binding to the SARS-CoV-2 spike (S) antigen was identified by a nonamplified biopanning method, which exhibited high affinity to the target, with a dissociation constant of 9.29 ± 1.55 nM. Molecular docking analysis reveals that this peptide binds to the N-terminal domain of the SARS-CoV-2 S antigen. Then, using this peptide as a capture probe and angiotensin-converting enzyme 2 as a detection probe, a peptide-based lateral flow immunoassay (pLFIA) for the sensitive detection of the SARS-CoV-2 S antigen without any antibody was developed, for which a polydopamine nanosphere (PDA)@MnO 2 nanocomposite with excellent oxidase-like activity was used as a colorimetric label, exhibiting dual-mode remarkable signal amplification of natural melanin and on-demand nanozyme catalytic enhancement. The PDA@MnO 2 -based pLFIA is capable of detecting the SARS-CoV-2 S antigen with a limit of detection of 8.01 pg/mL, which is 18.7 times lower than that of a conventional pLFIA tagged with gold nanoparticles. Additionally, the as-proposed PDA@MnO 2 -based pLFIA can detect up to 150 transduction units/mL SARS-CoV-2 pseudoviruses spiked in saliva samples. Given the outstanding analytical performance, the proposed PDA@MnO 2 -based pLFIA may offer a reliable option for the rapid diagnosis of SARS-CoV-2.

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Development and Analytical Evaluation of a Point-of-Care Electrochemical Biosensor for Rapid and Accurate SARS-CoV-2 Detection

Meshesha

Sardar

Supekar

et al. 2023

Preprint

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The COVID-19 pandemic has underscored the critical need for rapid and accurate screening and diagnostic methods for potential respiratory viruses. Existing COVID-19 diagnostic approaches face limitations either in terms of turnaround time or accuracy. In this study, we present an electrochemical biosensor that offers nearly instantaneous and precise SARS-CoV-2 detection, suitable for point-of-care and environmental monitoring applications. The biosensor employs a stapled hACE-2 N-terminal alpha helix peptide to functionalize an in-situ grown polypyrrole conductive polymer on a nitrocellulose membrane backbone through a chemical process. We assessed the biosensor’s analytical performance using heat-inactivated omicron and delta variants of the SARS-CoV-2 virus in artificial saliva (AS) and nasal swabs (NS) samples diluted in a strong ionic solution. Virus identification was achieved through electrochemical impedance spectroscopy (EIS) and frequency analyses. The assay demonstrated a limit of detection of 40 TCID50/mL, with 95% sensitivity and 100% specificity. Notably, the biosensor exhibited no cross-reactivity when tested against the influenza virus. The entire testing process using the biosensor takes less than a minute. In summary, our biosensor exhibits promising potential in the battle against pandemic respiratory viruses, offering a platform for the creation of rapid, compact, portable, and point-of-care devices capable of multiplexing various viruses. This groundbreaking development has the capacity to significantly bolster our readiness and response to future viral outbreaks.

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Verification of lateral flow antigen tests for SARS-CoV-2 by qPCR directly from the test device

Cited by 5 publications

References 11 publications

Development and Analytical Evaluation of a Point-of-Care Electrochemical Biosensor for Rapid and Accurate SARS-CoV-2 Detection

Development and Analytical Evaluation of a Point-of-Care Electrochemical Biosensor for Rapid and Accurate SARS-CoV-2 Detection

Sensitive Lateral Flow Immunoassay Based on Specific Peptide and Superior Oxidase Mimics with a Universal Dual-Mode Significant Signal Amplification

Development and Analytical Evaluation of a Point-of-Care Electrochemical Biosensor for Rapid and Accurate SARS-CoV-2 Detection

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