Commercial Strip‐Inspired One‐Pot CRISPR‐Based Chip for Multiplexed Detection of Respiratory Viruses

Small Methods, Journal Year: 2024, Volume and Issue: 9(1)

Published: Sept. 19, 2024

The absence of sensitive, multiplexed, and point-of-care assays poses a critical obstacle in promptly responding to emerging human respiratory virus (HRV) pandemics. Herein, RECOGNIZER (re-building commercial pregnancy strips via large-size nanoflowers), an innovative one-pot CRISPR assay, is presented that employs commercially available identify several types HRVs. superiority the assay mainly relies on two aspects: (i) DNA nanoflowers possessing high surface-to-volume ratio well-defined surface allow for considerable probe loading density minimized non-specific interaction, achieving impressive signal-to-noise proportion exceeding tenfold at 1 nM target. (ii) design reaction, multi-channel chip, custom-made app enables rapid, sample-to-answer, multiplexed analysis four HRVs 25 min. This demonstrates sensitivity 5.42 pM synthetic SARS-CoV-2 RNA 10 copies µL-1 plasmids after pre-amplification. Finally, proposed approach indicated 100% accuracy 50 clinical swab samples, demonstrating robust performance distinguishing from other versatility scalability renders it user-friendly platform infection monitoring, offering significant potential improving pandemic response efforts.

Language: Английский

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Nanoplasmonic microarray–based solid-phase amplification for highly sensitive and multiplexed molecular diagnostics: application for detecting SARS-CoV-2

Microchimica Acta, Journal Year: 2024, Volume and Issue: 191(11)

Published: Oct. 29, 2024

A novel approach is introduced using nanoplasmonic microarray–based solid-phase recombinase polymerase amplification (RPA) that offers high sensitivity and multiplexing capabilities for gene detection. Nanoplasmonic microarrays were developed through one-step immobilization of streptavidin/biotin primers fine-tuning the amplicon size to achieve plasmon-enhanced fluorescence (PEF) on substrate, thereby improving sensitivity. The specificity RPA was evaluated in detecting E, N, RdRP genes SARS-CoV-2. High achieved by minimizing primer-dimer formation employing a stringent washing process obtained with limit detection four copies per reaction within 30 min. In clinical testing nasopharyngeal swab samples (n = 30), demonstrated 100% consistency PCR results SARS-CoV-2, including differentiation Omicron mutations BA.1 BA.2. This overcomes issue amplification, as well rapidity, capabilities, simplified equipment isothermal reaction, making it valuable tool on-site molecular diagnostics.

Language: Английский

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State-of-the-Art Signal Amplification Strategies for Nucleic Acid and Non-Nucleic Acid Biosensors

Sensors and Actuators Reports, Journal Year: 2024, Volume and Issue: unknown, P. 100268 - 100268

Published: Dec. 1, 2024

Language: Английский

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Multiplexed discrimination of SARS-CoV-2 variants via duplex-specific nuclease combined MALDI-TOF MS

Analytical and Bioanalytical Chemistry, Journal Year: 2024, Volume and Issue: 416(8), P. 1833 - 1842

Published: Feb. 17, 2024

The frequent mutations in SARS-CoV-2 significantly increase the virus's pathogenicity and transmissibility while also diminishing effectiveness of vaccines. Consequently, assays capable rapidly simultaneously identifying multiple variants are essential for large-scale applications that aim to monitor evolution virus. In this work, we propose a method combining duplex-specific nuclease (DSN)-assisted cyclic amplification with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) detection, enabling simultaneous identification at high-throughput. Due high specificity DSN, single-base can be resolved by method. With ultra-sensitive detection MALDI-TOF MS, limit 100 pM viral RNA fragment was demonstrated. assay used typing Alpha, Beta, Delta variants. whole accomplished within 3 h, is performed under constant temperature, making technique simple operation efficient. It feasible extend many other We expect add value rapid screening play an important role pandemic control.

Language: Английский

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Deciphering ACE2-RBD binding affinity through peptide scanning: A molecular dynamics simulation approach

Computers in Biology and Medicine, Journal Year: 2024, Volume and Issue: 173, P. 108325 - 108325

Published: March 16, 2024

Language: Английский

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