Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees

Autores
Lannutti, Lucas; Mira, Anabela; Basualdo, Marina; Rodriguez, Graciela Adriana; Erler, Silvio; Silva, Victoria Ayelén; Gisder, Sebastian; Genersch, Elke; Florin-Christensen, Mónica; Schnittger, Leonhard
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Nosema ceranae is a ubiquitous microsporidian pathogen infecting the midgut of honey bees. The infection causes bee nosemosis, a disease associated with malnutrition, dysentery, and lethargic behavior, and results in considerable economic losses in apiculture. The use of a rapid, sensitive, and inexpensive DNA-based molecular detection method assists in the surveillance and eventual control of this pathogen. To this end, a loop-mediated isothermal amplification (LAMP) assay targeting the single-copy gene encoding the polar tube protein 3 (PTP3) has been developed. Genomic DNA of N. ceranae–infected forager bees sampled from distant geographic regions could be reliably amplified using the established LAMP assay. The N. ceranae-LAMP showed higher sensitivity than a classical reference PCR (98.6 vs 95.7%), when both approaches were applied to the detection of N. ceranae. LAMP detected a ten-fold lower infection rate than the reference PCR (1 pg vs 10 pg genomic DNA, respectively). In addition, we show highly specific and sensitive detection of N. ceranae from spore preparations in a direct LAMP format. No cross-reactions with genomic DNA and/or spores from N. apis, often co-infecting A. mellifera, or from N. bombi, infecting bumble bees, were observed. This low-cost and time-saving molecular detection method can be easily applied in simple laboratory settings, facilitating a rapid detection of N. ceranae in honey bees in epidemiological studies, surveillance and control, as well as evaluation of therapeutic measures against nosemosis.
Instituto de Patobiología
Fil: Lannutti, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Lannutti, Lucas. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; Argentina
Fil: Mira, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina.
Fil: Basualdo, Marina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. PROANVET; Argentina
Fil: Rodriguez, Graciela Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; Argentina
Fil: Erler, Silvio. Martin-Luther-Universität Halle-Wittenberg. Institut für Biologie, Molekulare Ökologie; Alemania
Fil: Erler, Silvio. Federal Research Centre for Cultivated Plants. Julius Kühn-Institut. Institute for Bee Protection; Alemania
Fil: Silva, Victoria Ayelén. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Gisder, Sebastian. Institute for Bee Research. Department of Molecular Microbiology and Bee Diseases; Alemania
Fil: Genersch, Elke. Institute for Bee Research. Department of Molecular Microbiology and Bee Diseases; Alemania
Fil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina
Fil: Florin-Christensen, Monica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Florin-Christensen, Monica. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; Argentina
Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina
Fil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Schnittger, Leonhard. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; Argentina
Fuente
Parasitology Research 119 : 3947-3956 (2020)
Materia
Nosema
Abeja Melífera
Parásitos
Apis mellifera
Nosema apis
Honey Bees
Parasites
Nosema ceranae
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/11124
Acceder
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oai_identifier_str oai:localhost:20.500.12123/11124
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network_name_str INTA Digital (INTA)
spelling Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey beesLannutti, LucasMira, AnabelaBasualdo, MarinaRodriguez, Graciela AdrianaErler, SilvioSilva, Victoria AyelénGisder, SebastianGenersch, ElkeFlorin-Christensen, MónicaSchnittger, LeonhardNosemaAbeja MelíferaParásitosApis melliferaNosema apisHoney BeesParasitesNosema ceranaeNosema ceranae is a ubiquitous microsporidian pathogen infecting the midgut of honey bees. The infection causes bee nosemosis, a disease associated with malnutrition, dysentery, and lethargic behavior, and results in considerable economic losses in apiculture. The use of a rapid, sensitive, and inexpensive DNA-based molecular detection method assists in the surveillance and eventual control of this pathogen. To this end, a loop-mediated isothermal amplification (LAMP) assay targeting the single-copy gene encoding the polar tube protein 3 (PTP3) has been developed. Genomic DNA of N. ceranae–infected forager bees sampled from distant geographic regions could be reliably amplified using the established LAMP assay. The N. ceranae-LAMP showed higher sensitivity than a classical reference PCR (98.6 vs 95.7%), when both approaches were applied to the detection of N. ceranae. LAMP detected a ten-fold lower infection rate than the reference PCR (1 pg vs 10 pg genomic DNA, respectively). In addition, we show highly specific and sensitive detection of N. ceranae from spore preparations in a direct LAMP format. No cross-reactions with genomic DNA and/or spores from N. apis, often co-infecting A. mellifera, or from N. bombi, infecting bumble bees, were observed. This low-cost and time-saving molecular detection method can be easily applied in simple laboratory settings, facilitating a rapid detection of N. ceranae in honey bees in epidemiological studies, surveillance and control, as well as evaluation of therapeutic measures against nosemosis.Instituto de PatobiologíaFil: Lannutti, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lannutti, Lucas. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; ArgentinaFil: Mira, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina.Fil: Basualdo, Marina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. PROANVET; ArgentinaFil: Rodriguez, Graciela Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; ArgentinaFil: Erler, Silvio. Martin-Luther-Universität Halle-Wittenberg. Institut für Biologie, Molekulare Ökologie; AlemaniaFil: Erler, Silvio. Federal Research Centre for Cultivated Plants. Julius Kühn-Institut. Institute for Bee Protection; AlemaniaFil: Silva, Victoria Ayelén. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Gisder, Sebastian. Institute for Bee Research. Department of Molecular Microbiology and Bee Diseases; AlemaniaFil: Genersch, Elke. Institute for Bee Research. Department of Molecular Microbiology and Bee Diseases; AlemaniaFil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; ArgentinaFil: Florin-Christensen, Monica. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Florin-Christensen, Monica. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; ArgentinaFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; ArgentinaFil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Schnittger, Leonhard. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; ArgentinaSpringer2022-01-14T12:46:17Z2022-01-14T12:46:17Z2020-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/11124https://link.springer.com/article/10.1007/s00436-020-06915-w0932-01131432-1955https://doi.org/10.1007/s00436-020-06915-wParasitology Research 119 : 3947-3956 (2020)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2019-PE-E1-I017-001/2019-PE-E1-I017-001/AR./DESARROLLO DEL SECTOR APÍCOLA ORGANIZADO, SUSTENTABLE Y COMPETITIVOinfo:eu-repo/semantics/restrictedAccess2025-10-16T09:30:21Zoai:localhost:20.500.12123/11124instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-10-16 09:30:22.656INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
title Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
spellingShingle Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
Lannutti, Lucas
Nosema
Abeja Melífera
Parásitos
Apis mellifera
Nosema apis
Honey Bees
Parasites
Nosema ceranae
title_short Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
title_full Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
title_fullStr Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
title_full_unstemmed Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
title_sort Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
dc.creator.none.fl_str_mv Lannutti, Lucas
Mira, Anabela
Basualdo, Marina
Rodriguez, Graciela Adriana
Erler, Silvio
Silva, Victoria Ayelén
Gisder, Sebastian
Genersch, Elke
Florin-Christensen, Mónica
Schnittger, Leonhard
author Lannutti, Lucas
author_facet Lannutti, Lucas
Mira, Anabela
Basualdo, Marina
Rodriguez, Graciela Adriana
Erler, Silvio
Silva, Victoria Ayelén
Gisder, Sebastian
Genersch, Elke
Florin-Christensen, Mónica
Schnittger, Leonhard
author_role author
author2 Mira, Anabela
Basualdo, Marina
Rodriguez, Graciela Adriana
Erler, Silvio
Silva, Victoria Ayelén
Gisder, Sebastian
Genersch, Elke
Florin-Christensen, Mónica
Schnittger, Leonhard
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Nosema
Abeja Melífera
Parásitos
Apis mellifera
Nosema apis
Honey Bees
Parasites
Nosema ceranae
topic Nosema
Abeja Melífera
Parásitos
Apis mellifera
Nosema apis
Honey Bees
Parasites
Nosema ceranae
dc.description.none.fl_txt_mv Nosema ceranae is a ubiquitous microsporidian pathogen infecting the midgut of honey bees. The infection causes bee nosemosis, a disease associated with malnutrition, dysentery, and lethargic behavior, and results in considerable economic losses in apiculture. The use of a rapid, sensitive, and inexpensive DNA-based molecular detection method assists in the surveillance and eventual control of this pathogen. To this end, a loop-mediated isothermal amplification (LAMP) assay targeting the single-copy gene encoding the polar tube protein 3 (PTP3) has been developed. Genomic DNA of N. ceranae–infected forager bees sampled from distant geographic regions could be reliably amplified using the established LAMP assay. The N. ceranae-LAMP showed higher sensitivity than a classical reference PCR (98.6 vs 95.7%), when both approaches were applied to the detection of N. ceranae. LAMP detected a ten-fold lower infection rate than the reference PCR (1 pg vs 10 pg genomic DNA, respectively). In addition, we show highly specific and sensitive detection of N. ceranae from spore preparations in a direct LAMP format. No cross-reactions with genomic DNA and/or spores from N. apis, often co-infecting A. mellifera, or from N. bombi, infecting bumble bees, were observed. This low-cost and time-saving molecular detection method can be easily applied in simple laboratory settings, facilitating a rapid detection of N. ceranae in honey bees in epidemiological studies, surveillance and control, as well as evaluation of therapeutic measures against nosemosis.
Instituto de Patobiología
Fil: Lannutti, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Lannutti, Lucas. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; Argentina
Fil: Mira, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina.
Fil: Basualdo, Marina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. PROANVET; Argentina
Fil: Rodriguez, Graciela Adriana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Hilario Ascasubi; Argentina
Fil: Erler, Silvio. Martin-Luther-Universität Halle-Wittenberg. Institut für Biologie, Molekulare Ökologie; Alemania
Fil: Erler, Silvio. Federal Research Centre for Cultivated Plants. Julius Kühn-Institut. Institute for Bee Protection; Alemania
Fil: Silva, Victoria Ayelén. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Gisder, Sebastian. Institute for Bee Research. Department of Molecular Microbiology and Bee Diseases; Alemania
Fil: Genersch, Elke. Institute for Bee Research. Department of Molecular Microbiology and Bee Diseases; Alemania
Fil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina
Fil: Florin-Christensen, Monica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Florin-Christensen, Monica. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; Argentina
Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina
Fil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Schnittger, Leonhard. Universidad de Morón. Escuela Superior de Ciencias Exactas y Naturales; Argentina
description Nosema ceranae is a ubiquitous microsporidian pathogen infecting the midgut of honey bees. The infection causes bee nosemosis, a disease associated with malnutrition, dysentery, and lethargic behavior, and results in considerable economic losses in apiculture. The use of a rapid, sensitive, and inexpensive DNA-based molecular detection method assists in the surveillance and eventual control of this pathogen. To this end, a loop-mediated isothermal amplification (LAMP) assay targeting the single-copy gene encoding the polar tube protein 3 (PTP3) has been developed. Genomic DNA of N. ceranae–infected forager bees sampled from distant geographic regions could be reliably amplified using the established LAMP assay. The N. ceranae-LAMP showed higher sensitivity than a classical reference PCR (98.6 vs 95.7%), when both approaches were applied to the detection of N. ceranae. LAMP detected a ten-fold lower infection rate than the reference PCR (1 pg vs 10 pg genomic DNA, respectively). In addition, we show highly specific and sensitive detection of N. ceranae from spore preparations in a direct LAMP format. No cross-reactions with genomic DNA and/or spores from N. apis, often co-infecting A. mellifera, or from N. bombi, infecting bumble bees, were observed. This low-cost and time-saving molecular detection method can be easily applied in simple laboratory settings, facilitating a rapid detection of N. ceranae in honey bees in epidemiological studies, surveillance and control, as well as evaluation of therapeutic measures against nosemosis.
publishDate 2020
dc.date.none.fl_str_mv 2020-10
2022-01-14T12:46:17Z
2022-01-14T12:46:17Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/11124
https://link.springer.com/article/10.1007/s00436-020-06915-w
0932-0113
1432-1955
https://doi.org/10.1007/s00436-020-06915-w
url http://hdl.handle.net/20.500.12123/11124
https://link.springer.com/article/10.1007/s00436-020-06915-w
https://doi.org/10.1007/s00436-020-06915-w
identifier_str_mv 0932-0113
1432-1955
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2019-PE-E1-I017-001/2019-PE-E1-I017-001/AR./DESARROLLO DEL SECTOR APÍCOLA ORGANIZADO, SUSTENTABLE Y COMPETITIVO
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv Parasitology Research 119 : 3947-3956 (2020)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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