Advancements and applications of loop-mediated isothermal amplification technology: a comprehensive overview

Frontiers in Microbiology, Journal Year: 2024, Volume and Issue: 15

Published: July 17, 2024

Loop-mediated isothermal amplification (LAMP) is a novel method for nucleic acid detection known its properties, high efficiency, sensitivity, and specificity. LAMP employs 4 to 6 primers targeting 8 regions of the desired sequence, allowing at temperatures between 60 65°C production up 10

Language: Английский

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Graphene Metasurface-Based Surface Plasmon Resonance Biosensor for Rapid COVID-19 Detection with Machine Learning Optimization

Plasmonics, Journal Year: 2025, Volume and Issue: unknown

Published: March 12, 2025

Language: Английский

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A Rapid PCR-LAMP Assay for the Early Detection of Lasiodiplodia theobromae from Basal Stem Rot-Infected Passion Fruit Plants

Molecular Biotechnology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 17, 2025

Language: Английский

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Rapid and visual identification of HIV-1 using reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticle-based lateral flow assay platform

Frontiers in Microbiology, Journal Year: 2023, Volume and Issue: 14

Published: July 12, 2023

Human immunodeficiency virus type one (HIV-1) infection remains a major public health problem worldwide. Early diagnosis of HIV-1 is crucial to treat and control this effectively. Here, for the first time, we reported novel molecular diagnostic assay called reverse transcription loop-mediated isothermal amplification combined with visual gold nanoparticle-based lateral flow (RT-LAMP-AuNPs-LFA), which devised rapid, specific, sensitive, identification HIV-1. The unique LAMP primers were successfully designed based on pol gene from genotypes CRF01_AE, CRF07_BC, CRF08_BC, subtype B, are prevalent in China. optimal HIV-1-RT-LAMP-AuNPs-LFA reaction conditions determined be 68°C 35 min. detection procedure, including crude genomic RNA isolation (approximately 5 min), RT-LAMP (35 result readout (<2 can completed within 45 Our has limit 20 copies per test, did not observe any cross-reactivity other pathogen our testing. Hence, preliminary results indicated that potentially serve as useful point-of-care tool clinical setting.

Language: Английский

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Development of LAMP assay for early detection of Yersinia ruckeri in aquaculture

PeerJ, Journal Year: 2025, Volume and Issue: 13, P. e19015 - e19015

Published: Feb. 25, 2025

Yersinia ruckeri is the causative agent of yersiniosis or enteric red mouth disease (ERM) that causes significant economic losses in salmonid aquaculture industry. Due to an increasing number outbreaks, lack effective vaccines and bacteria’s ability survive environment for long periods, there a necessity novel measures control ERM. New techniques capable rapidly detecting Y. are critical aid programs. Molecular methods, like real-time polymerase chain reaction, can detect ; however, methodology not field-deployable cannot support local decision-making during outbreak. We present molecular assay using loop mediated isothermal amplification (LAMP) water filtering method detection eDNA from samples improve current surveillance methods. The was optimised amplify glutamine synthetase gene ( glnA ) under 20 min. demonstrated high specificity sensitivity, as it did any non-target bacteria typically sources. It achieved limit (LOD) 0.5 × 10 −7 ng/µl, significantly surpassing LOD −4 ng/µl obtained through conventional reaction (cPCR). When applied environmental spiked with transformed Escherichia coli containing G-block target gene, Yr -LAMP exhibited analytical sensitivity 0.08 cells/µl initial filtered sample. Notably, cumulative time sample preparation 1 h. simplicity developed Yr- LAMP makes suitable routine procedure monitor fish ERM infection. This will enable informed on mitigating pathogen prevalence farms.

Language: Английский

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A new approach for the detection of genetic alterations utilizing modified loop-mediated isothermal amplification reaction (LAMP)

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: March 8, 2025

The increasing use of genetic testing for personalised therapy, highlights the need rapid, reliable diagnostics. Current methods are hindered by complex workflows, requiring advanced equipment, skilled personnel, and invasive tissue sampling. Loop-mediated isothermal amplification (LAMP) has emerged as a more efficient alternative to traditional PCR. LAMP eliminates thermal cycling, allowing faster, cost-effective tests, is less sensitive inhibitors, enabling from minimally processed samples. Although newer longer assay development time than PCR, its potential in oncology, particularly detecting changes, promising. We have developed LAMP-based method variations, optimized point-of-care testing. This technique uses modified primers with alterations at 3' end either F2 or B2 primers, ensuring specificity altered sequences. only produces positive signal when variant present, distinguishing it wild-type DNA. Our findings demonstrate that this high sensitivity, even samples both mutated material. Paired portable device, diagnostic could revolutionize alteration detection, offering quicker results improving treatment outcomes, targeted therapies.

Language: Английский

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Rapid Detection of Salmonella and Listeria monocytogenes in Fresh Beef Using Loop-Mediated Isothermal Amplification

Foodborne Pathogens and Disease, Journal Year: 2025, Volume and Issue: unknown

Published: April 28, 2025

This study developed and optimized loop-mediated isothermal amplification (LAMP) assays for rapid detection of Salmonella L. monocytogenes in fresh beef. LAMP primers targeting the invA gene hly were used. Reaction conditions including temperature, dNTP concentration, Mg2+ primer ratio, Bst DNA polymerase amount each pathogen. Differences observed between two pathogens optimal requirements, highlighting importance pathogen-specific optimization. The demonstrated high sensitivity with limits 120 fg/μL 130 monocytogenes, achieving within 40 60 min, respectively. Specificity tests confirmed both highly specific their target beef samples no cross-reactivity observed. Addition hydroxynaphthol blue enabled simple visual positive results, eliminating need specialized equipment. offer rapid, sensitive, these important foodborne pathogens.

Language: Английский

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Loop-Mediated Isothermal Amplification Assay for Visual Detection of Salmonella enterica Serovar Typhimurium in Food Animal Meat Products

Foods, Journal Year: 2025, Volume and Issue: 14(10), P. 1731 - 1731

Published: May 13, 2025

Detection of Salmonella, a highly diverse foodborne pathogen, is paramount to ensure safety and protection the animal industry its consumers. Salmonella enterica serovar Typhimurium among most important non-typhoidal serovars causing gastroenteritis worldwide. However, traditional identification labor- resource-intensive, while typical molecular tools require expensive reagents equipment. Hence, this study developed optimized calcein-based closed-tube loop-mediated isothermal amplification (LAMP)-based assay detect S. following enrichment steps compared with an PCR assay. The showed 100% specificity in silico confirmed through DNA sequencing. For actual testing, both LAMP Typhimurium. sensitivity, limit detection 22 pg/μL, 100-fold higher sensitivity at 220 fg/μL. Meanwhile, for pure culture assays detected least 4.98 × 104 CFU/mL. Parallel testing 208 raw meat samples from wet markets Metro Manila, Philippines, corroboration statistical association 89.42% 90.87% positivity rates Typhimurium, respectively. potentially powerful yet simple, sensitive, fast method detection.

Language: Английский

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Bringing the heat: Thermostable analogs of Bst polymerase allow high‐temperature LAMP

European Journal of Clinical Investigation, Journal Year: 2025, Volume and Issue: unknown

Published: May 13, 2025

Abstract Background Loop‐mediated isothermal amplification (LAMP) is a nucleic acid method that gained prominence during the early months of COVID‐19 pandemic due to its simplicity, sensitivity and robustness. However, this technique susceptible non‐specific amplifications, raising concerns about false‐positive results reduced diagnostic accuracy. A primary contributor testing primer dimerization, which can theoretically be mitigated by performing reactions at higher temperatures. Unfortunately, strand‐displacing DNA polymerases typically used in LAMP, such as Bst, exhibit efficiency elevated To address limitation, we hypothesised naturally occurring thermophilic analogs Bst may capable supporting LAMP temperatures, thereby improving reaction specificity. Methods Bioinformatics recombinant enzyme production allowed identification synthesis several analogs. These were tested real‐time assays detect diverse targets, wide range temperatures (63°C–75°C) presence typical qPCR inhibitors. Results Three polymerases—Bst_7, Bst_8 Bst_15—demonstrated exceptional activity robust stability temperature conditions (up 72.5°C), while displaying considerable resistance common Conclusions The identified represent potential solution for mitigation further boosting application molecular settings.

Language: Английский

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Double domain fusion improves the reverse transcriptase activity and inhibitor tolerance of Bst DNA polymerase

International Journal of Biological Macromolecules, Journal Year: 2024, Volume and Issue: 274, P. 133243 - 133243

Published: June 18, 2024

Language: Английский

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