Development of a loop-mediated isothermal amplification (LAMP) and a direct LAMP for the specific detection of Nosema ceranae, a parasite of honey bees
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 inex...
| Autores principales: | , , , , , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Springer
2022
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| Materias: | |
| Acceso en línea: | 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 |
| _version_ | 1855036514374778880 |
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| author | Lannutti, Lucas Mira, Anabela Basualdo, Marina Rodriguez, Graciela Adriana Erler, Silvio Silva, Victoria Ayelén Gisder, Sebastian Genersch, Elke Florin-Christensen, Mónica Schnittger, Leonhard |
| author_browse | Basualdo, Marina Erler, Silvio Florin-Christensen, Mónica Genersch, Elke Gisder, Sebastian Lannutti, Lucas Mira, Anabela Rodriguez, Graciela Adriana Schnittger, Leonhard Silva, Victoria Ayelén |
| 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_sort | Lannutti, Lucas |
| collection | INTA Digital |
| 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. |
| format | info:ar-repo/semantics/artículo |
| id | INTA11124 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA111242022-01-14T12:48:50Z 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 Mira, Anabela Basualdo, Marina Rodriguez, Graciela Adriana Erler, Silvio Silva, Victoria Ayelén Gisder, Sebastian Genersch, Elke Florin-Christensen, Mónica Schnittger, Leonhard Nosema Abeja Melífera Parásitos Apis mellifera Nosema apis Honey Bees Parasites Nosema ceranae 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 2022-01-14T12:46:17Z 2022-01-14T12:46:17Z 2020-10 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng info:eu-repograntAgreement/INTA/2019-PE-E1-I017-001/2019-PE-E1-I017-001/AR./DESARROLLO DEL SECTOR APÍCOLA ORGANIZADO, SUSTENTABLE Y COMPETITIVO info:eu-repo/semantics/restrictedAccess application/pdf Springer Parasitology Research 119 : 3947-3956 (2020) |
| spellingShingle | Nosema Abeja Melífera Parásitos Apis mellifera Nosema apis Honey Bees Parasites Nosema ceranae Lannutti, Lucas Mira, Anabela Basualdo, Marina Rodriguez, Graciela Adriana Erler, Silvio Silva, Victoria Ayelén Gisder, Sebastian Genersch, Elke Florin-Christensen, Mónica Schnittger, Leonhard 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 |
| 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_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_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 |
| topic | Nosema Abeja Melífera Parásitos Apis mellifera Nosema apis Honey Bees Parasites Nosema ceranae |
| 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 |
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