| Autor: |
Lannutti L; Escuela Superior de Ciencias Exactas y Naturales, Universidad de Morón, Morón, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina., Mira A; Instituto de Patobiología Veterinaria, CICVyA, INTA-Castelar, Los Reseros y Nicolas Repetto s/n, 1686 Hurlingham, Buenos Aires, Argentina., Basualdo M; Facultad de Ciencias Veterinarias, PROANVET, Universidad Nacional del Centro de la Provincia de Buenos Aires, Tandil, 7000, Buenos Aires, Argentina., Rodriguez G; Instituto Nacional de Tecnología Agropecuaria EEA Hilario Ascasubi, Hilario Ascasubi, Buenos Aires, Argentina., Erler S; Institut für Biologie, Molekulare Ökologie, Martin-Luther-Universität Halle-Wittenberg, 06099, Halle (Saale), Germany.; Institute for Bee Protection, Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, 38104, Braunschweig, Germany., Silva V; Instituto de Patobiología Veterinaria, CICVyA, INTA-Castelar, Los Reseros y Nicolas Repetto s/n, 1686 Hurlingham, Buenos Aires, Argentina., Gisder S; Department of Molecular Microbiology and Bee Diseases, Institute for Bee Research, Hohen Neuendorf, Germany., Genersch E; Department of Molecular Microbiology and Bee Diseases, Institute for Bee Research, Hohen Neuendorf, Germany., Florin-Christensen M; Escuela Superior de Ciencias Exactas y Naturales, Universidad de Morón, Morón, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.; Instituto de Patobiología Veterinaria, CICVyA, INTA-Castelar, Los Reseros y Nicolas Repetto s/n, 1686 Hurlingham, Buenos Aires, Argentina., Schnittger L; Escuela Superior de Ciencias Exactas y Naturales, Universidad de Morón, Morón, Argentina. schnittger.leonhard@inta.gob.ar.; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. schnittger.leonhard@inta.gob.ar.; Instituto de Patobiología Veterinaria, CICVyA, INTA-Castelar, Los Reseros y Nicolas Repetto s/n, 1686 Hurlingham, Buenos Aires, Argentina. schnittger.leonhard@inta.gob.ar. |
| Abstrakt: |
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. |
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