A Smart Single‐Loop‐Mediated Isothermal Amplification Facilitates Flexible SNP Probe Design for On‐Site Rapid Differentiation of SARS‐CoV‐2 Omicron Variants

Advanced Science, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Rapid on-site typing methods for SARS-CoV-2 variants of concern are crucial its effective surveillance and control. Herein, a smart single-loop-mediated isothermal amplification (ssLAMP) method with the absence an inner primer but addition swarm differentiation Omicron is developed. This unique design strategy offers greater flexibility in introducing single nucleotide polymorphism (SNP) identification probes enables multiple detection assays including BA.1, BA.2, BA.3, BA.4, BA.5. A 3D-printed portable dual fluorescence visualization device smartphone app developed to enable point-of-care testing. assay rapid (within 90 min), highly sensitive (100 copies/reaction), specific (identification SNP) SARA-CoV-2 variants. The ssLAMP identifies five BA.5-positive samples among 97 nasopharyngeal swab from clinic, 100% concordance rate Sanger sequencing. system expected be utilized on-site, specific, variants, great application potential pathogen genotyping, early cancer screening, other areas SNP mutation detection.

Language: Английский

Two‐Stage Mixed‐Dye‐Based Isothermal Amplification with Ribonuclease‐Cleavable Enhanced Probes for Dual‐Visualization Detection of SARS‐CoV‐2 Variants of Interest

Advanced Science, Journal Year: 2024, Volume and Issue: 11(29)

Published: June 3, 2024

Abstract Rapid and visual detection of SARS‐CoV‐2 variants is vital for timely assessment variant transmission in resource‐limited settings. Here, a closed‐tube, two‐stage, mixed‐dye‐based isothermal amplification method with ribonuclease‐cleavable enhanced probes (REP), termed REP‐TMAP, dual‐visualization including JN.1, BA.2, BA.4/5, Delta introduced. The first stage REP‐TMAP reverse transcription recombinase polymerase the second synergistically mediated by REP mixed dyes cresol red hydroxy naphthol blue. In reaction, color change under ambient light indicates infection, while fluorescence blue excitation specifies type. On detecting transcribed RNA spike gene, this assay rapid (within 40 min), highly sensitive (10–200 copies per reaction), specific (identification single‐base mutations). Furthermore, has been clinically validated to accurately detect BA.4/5 from 102 human oropharyngeal swabs. proposed therefore holds great potentials provide rapid, dual‐visualization, sensitive, specific, point‐of‐care beyond.

Language: Английский

Duplex in-one-tube detection of two important shrimp farming pathogens with color indication using a method combining RPA and PfAgo

Aquaculture, Journal Year: 2024, Volume and Issue: unknown, P. 741949 - 741949

Published: Nov. 1, 2024

Language: Английский