A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.

Croci, Carolina - Martínez de la Escalera, Gabriela - Scavone, Paola - Kruk, Carla - Segura, Angel - Piccini​, Claudia

Resumen:

One of the major concerns about cyanobacterial blooms is that some of them are capable of producing metabolites that are toxic to animals and humans (cyanotoxins). Thus, their presence in water affects human uses, from direct consumption, irrigation, aquaculture, to recreational uses. In Uruguay, the cyanobacteria most frequently associated with these events are species of the genus Microcystis, which includes toxic (carrying the mcyA-J cluster) and non-toxic populations. Since microscopy-based methods are not able to distinguish between them and analytic methods are timeconsuming, the development of early detection techniques to generate early warnings and avoid health risk is straightforward. In this work, we developed a method for monitoring toxic Microcystis quickly, at low cost and without specific or expensive equipment to read the results. A loop-mediated isothermal amplification protocol (LAMP) was designed using the mcyJ gene, which has been described not to suffer recombination and is present in single copy in toxic populations, as a target. The detection of mcyJ gene was achieved for natural samples by LAMP reaction and was confirmed by real-time quantitative PCR. The limit of detection showed that it was able to detect 28 pg/μL of DNA (corresponding to 3.65 x 104 toxic cells of Microcystis) in 19 min with 100% sensitivity. By extending the incubation time to 40 min, ca. 5000 toxic Microcystis cells could be detected but with lower sensitivity (< 80%). This method constitutes a promising tool for early warning that could be used to make monitoring and water management decisions.

Detalles Bibliográficos
2025
Agencia Nacional de Investigación e Innovación
cyanotoxins
mcyJ
LAMP
early warning
Ciencias Naturales y Exactas
Ciencias Biológicas
Bioquímica y Biología Molecular
Inglés
Instituto de Investigaciones Biológicas Clemente Estable
IIBCE en REDI
https://hdl.handle.net/20.500.12381/5360
Acceso abierto
Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND)
_version_ 1877653939640336384
author Croci, Carolina
author2 Martínez de la Escalera, Gabriela
Scavone, Paola
Kruk, Carla
Segura, Angel
Piccini​, Claudia
author2_role author
author
author
author
author
author_facet Croci, Carolina
Martínez de la Escalera, Gabriela
Scavone, Paola
Kruk, Carla
Segura, Angel
Piccini​, Claudia
author_role author
bitstream.checksum.fl_str_mv fcc8b2ddb26c820405a37f9e0cae7f18
becc2f3c5b9734666a7a45e974a17bf8
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
bitstream.url.fl_str_mv https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5360/2/license.txt
https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5360/1/Abstract%20ICHA.pdf
collection IIBCE en REDI
dc.creator.none.fl_str_mv Croci, Carolina
Martínez de la Escalera, Gabriela
Scavone, Paola
Kruk, Carla
Segura, Angel
Piccini​, Claudia
dc.date.accessioned.none.fl_str_mv 2026-01-12T18:30:26Z
dc.date.available.none.fl_str_mv 2026-01-12T18:30:26Z
dc.date.issued.none.fl_str_mv 2025-10
dc.description.abstract.none.fl_txt_mv One of the major concerns about cyanobacterial blooms is that some of them are capable of producing metabolites that are toxic to animals and humans (cyanotoxins). Thus, their presence in water affects human uses, from direct consumption, irrigation, aquaculture, to recreational uses. In Uruguay, the cyanobacteria most frequently associated with these events are species of the genus Microcystis, which includes toxic (carrying the mcyA-J cluster) and non-toxic populations. Since microscopy-based methods are not able to distinguish between them and analytic methods are timeconsuming, the development of early detection techniques to generate early warnings and avoid health risk is straightforward. In this work, we developed a method for monitoring toxic Microcystis quickly, at low cost and without specific or expensive equipment to read the results. A loop-mediated isothermal amplification protocol (LAMP) was designed using the mcyJ gene, which has been described not to suffer recombination and is present in single copy in toxic populations, as a target. The detection of mcyJ gene was achieved for natural samples by LAMP reaction and was confirmed by real-time quantitative PCR. The limit of detection showed that it was able to detect 28 pg/μL of DNA (corresponding to 3.65 x 104 toxic cells of Microcystis) in 19 min with 100% sensitivity. By extending the incubation time to 40 min, ca. 5000 toxic Microcystis cells could be detected but with lower sensitivity (< 80%). This method constitutes a promising tool for early warning that could be used to make monitoring and water management decisions.
dc.description.sponsorship.none.fl_txt_mv Agencia Nacional de Investigación e Innovación
dc.identifier.anii.es.fl_str_mv FMV_1_2021_1_166354
FCE_3_2022_1_172223
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12381/5360
dc.language.iso.none.fl_str_mv eng
dc.rights.*.fl_str_mv Acceso abierto
dc.rights.license.none.fl_str_mv Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.source.es.fl_str_mv International Conference on Harmful Algae (ICHA). Punta Arenas, Chile, 19-24 de octubre de 2025
dc.source.none.fl_str_mv reponame:IIBCE en REDI
instname:Instituto de Investigaciones Biológicas Clemente Estable
instacron:Instituto de Investigaciones Biológicas Clemente Estable
dc.subject.anii.none.fl_str_mv Ciencias Naturales y Exactas
Ciencias Biológicas
Bioquímica y Biología Molecular
dc.subject.es.fl_str_mv cyanotoxins
mcyJ
LAMP
early warning
dc.title.none.fl_str_mv A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
dc.type.es.fl_str_mv Documento de conferencia
dc.type.none.fl_str_mv info:eu-repo/semantics/conferenceObject
dc.type.version.es.fl_str_mv Publicado
dc.type.version.none.fl_str_mv info:eu-repo/semantics/publishedVersion
description One of the major concerns about cyanobacterial blooms is that some of them are capable of producing metabolites that are toxic to animals and humans (cyanotoxins). Thus, their presence in water affects human uses, from direct consumption, irrigation, aquaculture, to recreational uses. In Uruguay, the cyanobacteria most frequently associated with these events are species of the genus Microcystis, which includes toxic (carrying the mcyA-J cluster) and non-toxic populations. Since microscopy-based methods are not able to distinguish between them and analytic methods are timeconsuming, the development of early detection techniques to generate early warnings and avoid health risk is straightforward. In this work, we developed a method for monitoring toxic Microcystis quickly, at low cost and without specific or expensive equipment to read the results. A loop-mediated isothermal amplification protocol (LAMP) was designed using the mcyJ gene, which has been described not to suffer recombination and is present in single copy in toxic populations, as a target. The detection of mcyJ gene was achieved for natural samples by LAMP reaction and was confirmed by real-time quantitative PCR. The limit of detection showed that it was able to detect 28 pg/μL of DNA (corresponding to 3.65 x 104 toxic cells of Microcystis) in 19 min with 100% sensitivity. By extending the incubation time to 40 min, ca. 5000 toxic Microcystis cells could be detected but with lower sensitivity (< 80%). This method constitutes a promising tool for early warning that could be used to make monitoring and water management decisions.
eu_rights_str_mv openAccess
format conferenceObject
id IIBCE_7119eea112a818065146d2390152c4dd
identifier_str_mv FMV_1_2021_1_166354
FCE_3_2022_1_172223
instacron_str Instituto de Investigaciones Biológicas Clemente Estable
institution Instituto de Investigaciones Biológicas Clemente Estable
instname_str Instituto de Investigaciones Biológicas Clemente Estable
language eng
network_acronym_str IIBCE
network_name_str IIBCE en REDI
oai_identifier_str oai:redi.anii.org.uy:20.500.12381/5360
publishDate 2025
reponame_str IIBCE en REDI
repository.mail.fl_str_mv vcarballo@iibce.edu.uy
repository.name.fl_str_mv IIBCE en REDI - Instituto de Investigaciones Biológicas Clemente Estable
repository_id_str 9421_3
rights_invalid_str_mv Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND)
Acceso abierto
spelling Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional. (CC BY-NC-ND)Acceso abiertoinfo:eu-repo/semantics/openAccess2026-01-12T18:30:26Z2026-01-12T18:30:26Z2025-10https://hdl.handle.net/20.500.12381/5360FMV_1_2021_1_166354FCE_3_2022_1_172223One of the major concerns about cyanobacterial blooms is that some of them are capable of producing metabolites that are toxic to animals and humans (cyanotoxins). Thus, their presence in water affects human uses, from direct consumption, irrigation, aquaculture, to recreational uses. In Uruguay, the cyanobacteria most frequently associated with these events are species of the genus Microcystis, which includes toxic (carrying the mcyA-J cluster) and non-toxic populations. Since microscopy-based methods are not able to distinguish between them and analytic methods are timeconsuming, the development of early detection techniques to generate early warnings and avoid health risk is straightforward. In this work, we developed a method for monitoring toxic Microcystis quickly, at low cost and without specific or expensive equipment to read the results. A loop-mediated isothermal amplification protocol (LAMP) was designed using the mcyJ gene, which has been described not to suffer recombination and is present in single copy in toxic populations, as a target. The detection of mcyJ gene was achieved for natural samples by LAMP reaction and was confirmed by real-time quantitative PCR. The limit of detection showed that it was able to detect 28 pg/μL of DNA (corresponding to 3.65 x 104 toxic cells of Microcystis) in 19 min with 100% sensitivity. By extending the incubation time to 40 min, ca. 5000 toxic Microcystis cells could be detected but with lower sensitivity (< 80%). This method constitutes a promising tool for early warning that could be used to make monitoring and water management decisions.Agencia Nacional de Investigación e InnovaciónengInternational Conference on Harmful Algae (ICHA). Punta Arenas, Chile, 19-24 de octubre de 2025reponame:IIBCE en REDIinstname:Instituto de Investigaciones Biológicas Clemente Estableinstacron:Instituto de Investigaciones Biológicas Clemente EstablecyanotoxinsmcyJLAMPearly warningCiencias Naturales y ExactasCiencias BiológicasBioquímica y Biología MolecularA loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.Documento de conferenciaPublicadoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectInstituto de Investigaciones Biologicas Clemente Estable//Ciencias Naturales y Exactas/Ciencias Biológicas/Bioquímica y Biología MolecularCroci, CarolinaMartínez de la Escalera, GabrielaScavone, PaolaKruk, CarlaSegura, AngelPiccini​, ClaudiaLICENSElicense.txtlicense.txttext/plain; charset=utf-85151https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5360/2/license.txtfcc8b2ddb26c820405a37f9e0cae7f18MD52ORIGINALAbstract ICHA.pdfAbstract ICHA.pdfResumen presentado para participar del congreso ICHAapplication/pdf64422https://redi.anii.org.uy/jspui/bitstream/20.500.12381/5360/1/Abstract%20ICHA.pdfbecc2f3c5b9734666a7a45e974a17bf8MD5120.500.12381/53602026-01-12 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científico-tecnológicohttps://www.gub.uy/ministerio-educacion-cultura/iibcehttps://redi.anii.org.uy/oai/requestvcarballo@iibce.edu.uyUruguayopendoar:9421_32026-01-12T18:30:27IIBCE en REDI - Instituto de Investigaciones Biológicas Clemente Establefalse
spellingShingle A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
Croci, Carolina
cyanotoxins
mcyJ
LAMP
early warning
Ciencias Naturales y Exactas
Ciencias Biológicas
Bioquímica y Biología Molecular
status_str publishedVersion
title A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
title_full A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
title_fullStr A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
title_full_unstemmed A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
title_short A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
title_sort A loop-mediated isothermal amplification method (LAMP) for detecting toxic Microcystis.
topic cyanotoxins
mcyJ
LAMP
early warning
Ciencias Naturales y Exactas
Ciencias Biológicas
Bioquímica y Biología Molecular
url https://hdl.handle.net/20.500.12381/5360