Biomicrofluidics, Journal Year: 2022, Volume and Issue: 16(6)
Published: Nov. 2, 2022
First Page
Language: Английский
International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(5), P. 1806 - 1806
Published: Feb. 20, 2025
Resource-limited settings and supply chain difficulties faced throughout the COVID-19 pandemic prompted development of rapid alternative methods detecting SARS-CoV-2. These include reverse-transcription loop-mediated isothermal amplification (RT-LAMP), recombinase polymerase (RT-RPA), CRISPR-Cas12a fluorescence detection. We describe RT-LAMP, RT-RPA, assays for detection N E-gene amplicons SARS-CoV-2 optimization various assay components, including incubation temperatures, Cas12a enzymes, reporter molecules, use a lyophilized RT-LAMP master mix. also testing one-tube RT-LAMP-CRISPR-Cas12a assay. The showed promise in reducing hands-on time improving time-to-result. found no improvements sensitivity with but did achieve at lower copy number mix compared to liquid reagent (50 vs. 100 copies 20 min). When used detect presence RNA clinical saliva samples from 75 infected patients, discriminatory ability optimized RT-LAMP-CRISPR was be comparable RT-qPCR, minor reduction sensitivity.
Language: Английский
Citations
2
Foods, Journal Year: 2024, Volume and Issue: 13(11), P. 1635 - 1635
Published: May 24, 2024
Loop-mediated isothermal amplification, LAMP, is nowadays the most popular nucleic acid amplification technique, and as such, several commercial, ready-to-use master mixes have flourished. Unfortunately, independent studies to determine their performance are limited. The current study performed an evaluation of existing commercial LAMP WarmStart® Kit, LavaLAMP™ DNA Master Mix, Saphir Bst Turbo GreenMaster, OptiGene Fast Mix ISO-004, SynLAMP Mix. To reduce bias, three different genes, namely ttr (Salmonella spp.), rfbE (E. coli O157), hly (Listeria monocytogenes), were targeted. comparison was based on speed, with decreasing concentrations, effect five typical reaction additives (betaine, DMSO, pullulan, TMAC, GuHCl). Significant differences observed among mixes. provided fastest showed less detrimental effects associated supplements evaluated. Out chemicals tested, pullulan best results in terms speed. It noteworthy that impacted differently. Overall, provides insights into mixes, which can be value for scientific community better select appropriate reagents when developing new methods.
Language: Английский
Citations
4
Gaceta Médica Boliviana, Journal Year: 2025, Volume and Issue: 48(1), P. 7 - 13
Published: Jan. 15, 2025
Objetivos: actualmente, la técnica estándar de detección virus qRT-PCR, requiere personal capacitado, equipos especializados y reactivos relativamente costosos. La amplificación isotérmica mediada por bucle, LAMP, ha resurgido como una alternativa prometedora previamente durante pandemia COVID-19 su estudio tiene potencial para el control enfermedades infecciosas emergentes. Métodos: en este trabajo, presentamos un comparativo realizado SEDES Tarija octubre del 2020 donde se comparó muestras pacientes voluntarios utilizando RT-PCR LAMP hisopados nasofaríngeos saliva. Resultados: demostró concordancia 100% con PCR ARN nasofaríngeo, 93,5% Conclusiones: diagnóstico rápido práctico emergentes es crucial establecer estrategias eficaces que minimicen impacto económico a vez maximicen protección sobre sus ciudadanos. Nuestros resultados validan sensibilidad uso saliva menos invasiva. Este trabajo confirma validez fácil implementación áreas rurales recursos limitados virales
Citations
0
Elsevier eBooks, Journal Year: 2025, Volume and Issue: unknown, P. 209 - 223
Published: Jan. 1, 2025
Language: Английский
Citations
0
Aquaculture, Journal Year: 2025, Volume and Issue: unknown, P. 742483 - 742483
Published: March 1, 2025
Language: Английский
Citations
0
Microchemical Journal, Journal Year: 2025, Volume and Issue: unknown, P. 113724 - 113724
Published: April 1, 2025
Language: Английский
Citations
0
ACS Sensors, Journal Year: 2024, Volume and Issue: 9(10), P. 5541 - 5549
Published: Oct. 8, 2024
Isothermal nucleic acid amplification tests, NAATs, such as reverse transcription-loop-mediated isothermal (RT-LAMP), offer promising capabilities to perform real-time semiquantitative detection of viral pathogens. These tests provide rapid results, utilize simple instrumentation for single-temperature reactions, support efficient user workflows, and are suitable field use. Herein, we present a novel robust method monitoring HIV-1 RNA RT-LAMP utilizing implementation particle diffusometry (PD), diffusivity quantification technique using fluorescent particles, quantify concentration in nuclease-free water. We monitor changes diffusion dynamics 400 nm fluorescently labeled particles throughout the water, enabling measurement within 20 min concentrations low 25 virus per μL. Moreover, single-blind study, demonstrate by accurately determining initial an unknown 10% absolute error margin. results highlight potential PD readout quantifying via RT-LAMP, offering promise load HIV other chronic infections.
Language: Английский
Citations
2
Biosensors, Journal Year: 2022, Volume and Issue: 12(11), P. 978 - 978
Published: Nov. 7, 2022
The COVID-19 pandemic caused by SARS-CoV-2 has drawn attention to the need for fast and accurate diagnostic testing. Concerns from emerging variants other circulating respiratory viral pathogens further underscore importance of expanding testing multiplex detection, as single-plex may fail detect viruses, while sequencing can be too slow expensive a tool. As result, there have been significant advances in nucleic-acid-based virus testing, creating timely review. This review first introduces frequent nucleic acid targets tests, then proceeds comprehensive up-to-date overview assays that incorporate various detection reactions readout modalities. performances, advantages, disadvantages these are discussed, followed highlights platforms amenable point-of-care use. Finally, this points out remaining technical challenges shares perspectives on future research development. By examining state art synthesizing existing development provide useful resource facilitating ultimately combating COVID-19.
Language: Английский
Citations
10
BMC Microbiology, Journal Year: 2023, Volume and Issue: 23(1)
Published: March 7, 2023
Abstract Background In the absence of effective antiviral drugs or vaccines, early and accurate detection SARS-CoV-2 infection is essential to COVID-19 pandemic. This study developed evaluated a novel rapid One-Step LAMP assay directly detect RNA from nasopharyngeal (NP) swab samples patients with suspected living in deprived areas comparison Real-time PCR. Methods Two hundred fifty-four NP western Iran were tested by TaqMan RT-qPCR fast assays. Tenfold serial dilutions standard strain where viral copy number each dilution was previously determined using qPCR various templates used investigate analytical sensitivity specificity triplicate. Also, efficacy reliability method compared positive negative clinical samples. Results The results tests 131 (51.6%) 127 (50%) participants, respectively. Based on Cohen’s kappa coefficient (κ), agreement between two 97%, which statistically significant ( P < 0.001). limit for 1 × 10 copies per reaction less than an hour triplicates. Negative all non-SARS-CoV-2 represent 100% specificity. Conclusions showed that efficient consistent technique detecting among individuals due its simplicity, speed, low cost, sensitivity, Therefore, it has great potential as useful diagnostic tool disease epidemic control, timely treatment, public health protection, especially poor underdeveloped countries.
Language: Английский
Citations
4
Journal of Thrombosis and Haemostasis, Journal Year: 2024, Volume and Issue: 22(6), P. 1627 - 1639
Published: Feb. 19, 2024
Language: Английский
Citations
1