dc.contributor.author | Crego Vicente, Beatriz | |
dc.contributor.author | Fernández Soto, Pedro | |
dc.contributor.author | García-Bernalt Diego, Juan | |
dc.contributor.author | Febrer Sendra, Begoña | |
dc.contributor.author | Muro Álvarez, Antonio | |
dc.date.accessioned | 2024-01-24T12:39:27Z | |
dc.date.available | 2024-01-24T12:39:27Z | |
dc.date.issued | 2023-01-03 | |
dc.identifier.citation | Crego-Vicente, B., Fernández-Soto, P., García-Bernalt Diego, J., Febrer-Sendra, B., & Muro, A. (2023). Development of a Duplex LAMP Assay with Probe-Based Readout for Simultaneous Real-Time Detection of Schistosoma mansoni and Strongyloides spp.-A Laboratory Approach to Point-Of-Care. International Journal of Molecular Sciences, 24(1), 893. | es_ES |
dc.identifier.issn | 1661-6596 | |
dc.identifier.uri | http://hdl.handle.net/10366/154618 | |
dc.description.abstract | [ENG] isothermal amplification (LAMP) is the most popular technology for point-of-care testing applications due its rapid, sensitive and specific detection with simple instrumentation compared to PCR-based methods. Many systems for reading the results of LAMP amplifications exist, including real-time fluorescence detection using fluorophore-labelled probes attached to oligonucleotide sequences complementary to the target nucleic acid. This methodology allows the simultaneous detection of multiple targets (multiplexing) in one LAMP assay. A method for multiplexing LAMP is the amplification by release of quenching (DARQ) technique by using a 5′-quencher modified LAMP primer annealed to 3′-fluorophore-labelled acting as detection oligonucleotide. The main application of multiplex LAMP is the rapid and accurate diagnosis of infectious diseases, allowing differentiation of co-infecting pathogens in a single reaction. Schistosomiasis, caused among other species by Schistosoma mansoni and strongyloidiasis, caused by Strongyloides stercoralis, are the most common helminth-parasite infections worldwide with overlapping distribution areas and high possibility of coinfections in the human population. It would be of great interest to develop a duplex LAMP to detect both pathogens in the same reaction. In this study, we investigate the use of our two previously developed and well-stablished LAMP assays for S. mansoni and Strongyloides spp. DNA detection in a new duplex real-time eight-primer system based on a modified DARQ probe method that can be performed in a portable isothermal fluorimeter with minimal laboratory resources. We also applied a strategy to stabilize the duplexed DARQ-LAMP mixtures at room temperature for use as ready-to-use formats facilitating analysis in field settings as point-of-care diagnostics for schistosomiasis and strongyloidiasis. | es_ES |
dc.format.mimetype | application/pdf | |
dc.language.iso | eng | es_ES |
dc.publisher | MDPI | es_ES |
dc.rights | CC0 1.0 Universal | * |
dc.rights.uri | http://creativecommons.org/publicdomain/zero/1.0/ | * |
dc.subject | Loop-mediated isothermal amplification | es_ES |
dc.subject | Schistosoma mansoni | es_ES |
dc.subject | Strongyloides | es_ES |
dc.subject | Point-of-care | es_ES |
dc.subject | Duplex LAMP assay | es_ES |
dc.subject | Multiplex LAMP | es_ES |
dc.subject.mesh | Schistosomiasis mansoni | * |
dc.subject.mesh | Point-of-Care Systems | * |
dc.subject.mesh | Schistosoma mansoni | * |
dc.subject.mesh | Molecular Diagnostic Techniques | * |
dc.subject.mesh | Strongyloides | * |
dc.title | Development of a Duplex LAMP Assay with Probe-Based Readout for Simultaneous Real-Time Detection of Schistosoma mansoni and Strongyloides spp. -A Laboratory Approach to Point-Of-Care | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.relation.publishversion | https://doi.org/10.3390/ijms24010893 | es_ES |
dc.subject.unesco | 24 Ciencias de la vida | es_ES |
dc.subject.unesco | 3207.12 Parasitología | es_ES |
dc.identifier.doi | 10.3390/ijms24010893 | |
dc.relation.projectID | This work was funded by the Institute of Health Carlos III, ISCIII, through the project PI19/01727 and cofunded by the European Union | es_ES |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | es_ES |
dc.identifier.essn | 1422-0067 | |
dc.journal.title | International Journal of Molecular Sciences | es_ES |
dc.volume.number | 24 | es_ES |
dc.issue.number | 1 | es_ES |
dc.page.initial | 893 | es_ES |
dc.type.hasVersion | info:eu-repo/semantics/publishedVersion | es_ES |
dc.subject.decs | Schistosoma mansoni | * |
dc.subject.decs | Strongyloides | * |
dc.subject.decs | esquistosomiasis intestinal | * |
dc.subject.decs | técnicas de diagnóstico molecular | * |
dc.subject.decs | sistemas de atención en la cabecera del paciente | * |