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